动态高压微射流技术制备中链脂肪酸纳米脂质体及其性质的研究
|
摘要
中链脂肪酸(MCFAs)具有吸收快、易供能和抑制体内脂肪蓄积等优点,但过量摄入会引发恶心、刺激胆囊素分泌和胃肠道不适等状况。脂质体作为一种优良的可生物降解性载体,可以提高药物的生物利用率。
本课题以MCFAs为油溶性模型材料,采用动态高压微射流(DHPM)作为核心技术,结合薄膜分散法和冻融法,通过工艺优化,制备薄膜分散-DHPM法的MCFAs纳米脂质体Ⅰ和DHPM-冻融法的MCFAs纳米脂质体Ⅱ,并且研究了这两种脂质体的物理化学性(粒径,多分散系数,zeta电位,包封率,载药量,外观形貌,DSC,渗漏率,表面张力,长期贮藏稳定性)和生理功能性(易供能性和抑制体内脂肪蓄积性)。
本课题建立了MCFAs体外分析方法：得到MCFAs含量为95.94%,辛酸与癸酸的质量比为5：4；确定了MCFAs脂质体包封率的测定方法为离心-正已烷萃取分离游离药物、碱法甲酯化MCFAs、气相色谱测定包封率。
制备了MCFAs纳米脂质体Ⅰ和Ⅱ,研究了各个配方因素和工艺因素,结果表明,MCFAs纳米脂质体Ⅰ的最优配方和工艺为：脂质浓度是8%, MCFAs浓度为15mg/mL,磷脂与胆固醇质量比为6：1,吐温-80浓度为30%,PBS浓度是0.05mol/L,PBS的pH值为7.4,脂质体的制备温度为40℃,处理压力是140MPa,处理次数为4次。MCFAs纳米脂质体Ⅱ的最优配方和工艺为：冷冻保护剂为麦芽糖,冷冻保护剂用量为3%,冻融次数为3次,冷冻时间为0.5h,融化温度为40℃。两种方法的精密度均较高。
对MCFAs纳米脂质体Ⅰ和Ⅱ进行了性质表征和质量评价,结果表明,MCFAs脂质体Ⅰ较好：外观形貌呈球形或椭球形,粒度为76.2±1.2nm, zeta电位为-50.51±0.4mV,分散系数为0.207±0.003,EE为70.5±6.5%,载药量为9.4±0.4%,表面张力为33.86±0.36dyne/cm,相变温度为138℃,渗透率低、长期贮藏稳定性好。
研究了MCFAs纳米脂质体Ⅰ和Ⅱ的易供能性。结果表明：用MCFAs纳米脂质体Ⅰ和Ⅱ灌胃小鼠,相比正常组,两种脂质体的高剂量组均可以显著提高小鼠负重游泳时间(分别为104±29min和105±31min)和肝糖原浓度(分别为16.40±1.45mg/g和14.98±3.56mg/g),可明显降低血清尿素氮(分别为891.5±113.4mg/L和839.5±111.9mg/L)和血乳酸(分别为6.05±1.40mmol/L和5.70±1.02mmol/L);另外,MCFAs组与两种MCFAs纳米脂质体组间没有显著性差异。
研究了MCFAs纳米脂质体Ⅰ和Ⅱ的抑制体内脂肪蓄积性。结果表明：用MCFAs纳米脂质体Ⅰ和Ⅱ灌胃小鼠两周(短期)和五周(长期),与正常组相比,两种脂质体对小鼠体重、粮食摄入量、脂肪重量和肝脏重量均无显著差异,但对TG(分别为0.550±0.140mmol/L和0.384±0.150mmol/L)、TCH(分别为0.621±0.097和0.540±0.247)有显著性差异；另外,MCFAs组与两种MCFAs纳米脂质体组间并无显著性差异。
总之,用薄膜分散-DHPM法和DHPM-冻融法制备的两种MCFAs纳米脂质体物理化学性状稳定,既克服了MCFAs自身的弱点,易供能性和抑制体内脂肪积蓄性又与单独使用MCFAs时没有显著性差异,可作为一种潜在的易供能产品和治疗肥胖症的膳食营养素。
Medium-chain fatty acids (MCFAs) possess the advantages of fast absorption, easy-energy supply and suppression of body fat accumulation. However, an excess intake of MCFAs can lead to nausea, stimulation to the secretion of cholecystokinin and gastrointestinal discomfort. As a promising biodegraded delivery system, liposomes can improve the bioavailability of drugs.
MCFAs were used as modeling materials in this study and incorporated into nanoliposomes. As the main technology, dynamic high pressure microfluidization (DHPM) was combined with thin-layer dispersion and freeze-thawing methods to prepared MCFAs nanoliposomes. In addition, the physical-chemical (size, polydisperse index, zeta potential, entrapment efficiency, drug loading, morphology, DSC, leakage rate, surface tension, long-time storage stability) and biological (easy-energy supply and suppression body fat accumulation) properties were investigated.
The analysis method of MCFAs in vitro was established, with the content of MCFAs 95.94% and the mass ratio between octoic acid and decoic acid 5/4. Besides, the percentage of MCFAs incorporated into liposomes was measured by gas chromatography after free drugs had been separated from liposomes by centrifugation, alkaline process methyl esterification of MCFAs and N-hexane extraction.
Thin-layer dispersion-DHPM and DHPM-freeze-thawing methods were developed to prepare MCFAs nanoliposomesⅠandⅡ, respectively. Each factor was analyzed and the orthogonal test was taken to optimize the formula. The optimized formula of MCFAs nanoliposomesⅠas follows:the concentration of lipids and MCFAs were 8% and 15 mg/mL, respectively; the mass ratio between phospholipid and cholesterol was 6/1;the concentration of tween-80 was 30%; the concentration and pH of PBS were 0.05 mol/L and 7.4, respectively; the preparation temperature was 40℃; the treatment pressure and passes were 140 MPa and 4, respectively. The optimized formula of MCFAs nanoliposomesⅡas following: cryoprotectant was maltose; the dosage of cryoprotectant was 3%; the freeze-thawing cycles were 3; the freezing time was 0.5 h and thawing temperature was 40℃. Both of the precision of the two methods were good.
The characteristics and qualities of both MCFAs nanoliposomesⅠandⅡwere evaluated. Compared with MCFAs nanoliposomesⅡ, MCFAs nanoliposomesⅠexhibited great advantages in their characteristics with regular spherical or elliptical in shape, a small average diameter (76.2±1.2 nm), narrow size distribution (PDI=0.207±0.003), high zeta potential (-50.51±0.4 mV), large entrapment efficiency (70.5±6.5%) and drug loading (9.4±0.4%), low surface tension (33.86±0.36 dyne/cm), high Tm (138℃), low leakage rate and good long-time storage stability.
The easy-energy-supply property of these two kinds of MCFAs nanoliposomes were estimated by the relive-fatigue experiments to the mice. The high-dose groups ofⅠandⅡ(680 mg/kg) suggested a significantly longer weight-loaded swimming time (104±29 min and 105±31min, respectively) and a higher hepatic glycogen (HG) (16.40±1.45 mg/g and 14.98±3.56 mg/g, respectively) than the control group. Moreover, the serum urea nitrogen (SUN) (891.5±113.4mg/L and 839.5±111.9mg/L, respectively) and blood lactic acid (LD) (6.05±1.40mmol/L and 5.70±1.02mmol/L, respectively) of high-dose groups were also significantly lower than those of the control group. However, the MCFAs group and the two MCFAs nanoliposomes groups showed no significant difference.
The suppression accumulation of body fat property of MCFAs nanoliposomesⅠandⅡwas investigated by short-term (two weeks) and long-term (five weeks) oral administration to the mice. Compared to the control group, the body weight of the mice, food intake, the adipose tissue and liver weights of the two MCFAs nanoliposomes were no significant difference. Nevertheless, the triglyceride (0.550±0.140 mmol/L and 0.384±0.150 mmol/L, respectively) and total cholesterol (0.621±0.097 and 0.540±0.247, respectively) were significantly lower than the control group. The MCFAs group and the tow MCFAs nanoliposomes groups showed no significant difference.
All the results indicated that MCFAs nanoliposomes prepared by thin-layer dispersion-DHPM and freeze-thawing-DHPM methods have overcome the drawbacks of MCFAs and exhibited good physicochemical properties. The easy-energy-supply and suppression of body fat accumulation characteristics revealed the MCFAs nanoliposomes can be a potential easy-energy-supply drug candidate.
本课题以MCFAs为油溶性模型材料,采用动态高压微射流(DHPM)作为核心技术,结合薄膜分散法和冻融法,通过工艺优化,制备薄膜分散-DHPM法的MCFAs纳米脂质体Ⅰ和DHPM-冻融法的MCFAs纳米脂质体Ⅱ,并且研究了这两种脂质体的物理化学性(粒径,多分散系数,zeta电位,包封率,载药量,外观形貌,DSC,渗漏率,表面张力,长期贮藏稳定性)和生理功能性(易供能性和抑制体内脂肪蓄积性)。
本课题建立了MCFAs体外分析方法：得到MCFAs含量为95.94%,辛酸与癸酸的质量比为5：4；确定了MCFAs脂质体包封率的测定方法为离心-正已烷萃取分离游离药物、碱法甲酯化MCFAs、气相色谱测定包封率。
制备了MCFAs纳米脂质体Ⅰ和Ⅱ,研究了各个配方因素和工艺因素,结果表明,MCFAs纳米脂质体Ⅰ的最优配方和工艺为：脂质浓度是8%, MCFAs浓度为15mg/mL,磷脂与胆固醇质量比为6：1,吐温-80浓度为30%,PBS浓度是0.05mol/L,PBS的pH值为7.4,脂质体的制备温度为40℃,处理压力是140MPa,处理次数为4次。MCFAs纳米脂质体Ⅱ的最优配方和工艺为：冷冻保护剂为麦芽糖,冷冻保护剂用量为3%,冻融次数为3次,冷冻时间为0.5h,融化温度为40℃。两种方法的精密度均较高。
对MCFAs纳米脂质体Ⅰ和Ⅱ进行了性质表征和质量评价,结果表明,MCFAs脂质体Ⅰ较好：外观形貌呈球形或椭球形,粒度为76.2±1.2nm, zeta电位为-50.51±0.4mV,分散系数为0.207±0.003,EE为70.5±6.5%,载药量为9.4±0.4%,表面张力为33.86±0.36dyne/cm,相变温度为138℃,渗透率低、长期贮藏稳定性好。
研究了MCFAs纳米脂质体Ⅰ和Ⅱ的易供能性。结果表明：用MCFAs纳米脂质体Ⅰ和Ⅱ灌胃小鼠,相比正常组,两种脂质体的高剂量组均可以显著提高小鼠负重游泳时间(分别为104±29min和105±31min)和肝糖原浓度(分别为16.40±1.45mg/g和14.98±3.56mg/g),可明显降低血清尿素氮(分别为891.5±113.4mg/L和839.5±111.9mg/L)和血乳酸(分别为6.05±1.40mmol/L和5.70±1.02mmol/L);另外,MCFAs组与两种MCFAs纳米脂质体组间没有显著性差异。
研究了MCFAs纳米脂质体Ⅰ和Ⅱ的抑制体内脂肪蓄积性。结果表明：用MCFAs纳米脂质体Ⅰ和Ⅱ灌胃小鼠两周(短期)和五周(长期),与正常组相比,两种脂质体对小鼠体重、粮食摄入量、脂肪重量和肝脏重量均无显著差异,但对TG(分别为0.550±0.140mmol/L和0.384±0.150mmol/L)、TCH(分别为0.621±0.097和0.540±0.247)有显著性差异；另外,MCFAs组与两种MCFAs纳米脂质体组间并无显著性差异。
总之,用薄膜分散-DHPM法和DHPM-冻融法制备的两种MCFAs纳米脂质体物理化学性状稳定,既克服了MCFAs自身的弱点,易供能性和抑制体内脂肪积蓄性又与单独使用MCFAs时没有显著性差异,可作为一种潜在的易供能产品和治疗肥胖症的膳食营养素。
Medium-chain fatty acids (MCFAs) possess the advantages of fast absorption, easy-energy supply and suppression of body fat accumulation. However, an excess intake of MCFAs can lead to nausea, stimulation to the secretion of cholecystokinin and gastrointestinal discomfort. As a promising biodegraded delivery system, liposomes can improve the bioavailability of drugs.
MCFAs were used as modeling materials in this study and incorporated into nanoliposomes. As the main technology, dynamic high pressure microfluidization (DHPM) was combined with thin-layer dispersion and freeze-thawing methods to prepared MCFAs nanoliposomes. In addition, the physical-chemical (size, polydisperse index, zeta potential, entrapment efficiency, drug loading, morphology, DSC, leakage rate, surface tension, long-time storage stability) and biological (easy-energy supply and suppression body fat accumulation) properties were investigated.
The analysis method of MCFAs in vitro was established, with the content of MCFAs 95.94% and the mass ratio between octoic acid and decoic acid 5/4. Besides, the percentage of MCFAs incorporated into liposomes was measured by gas chromatography after free drugs had been separated from liposomes by centrifugation, alkaline process methyl esterification of MCFAs and N-hexane extraction.
Thin-layer dispersion-DHPM and DHPM-freeze-thawing methods were developed to prepare MCFAs nanoliposomesⅠandⅡ, respectively. Each factor was analyzed and the orthogonal test was taken to optimize the formula. The optimized formula of MCFAs nanoliposomesⅠas follows:the concentration of lipids and MCFAs were 8% and 15 mg/mL, respectively; the mass ratio between phospholipid and cholesterol was 6/1;the concentration of tween-80 was 30%; the concentration and pH of PBS were 0.05 mol/L and 7.4, respectively; the preparation temperature was 40℃; the treatment pressure and passes were 140 MPa and 4, respectively. The optimized formula of MCFAs nanoliposomesⅡas following: cryoprotectant was maltose; the dosage of cryoprotectant was 3%; the freeze-thawing cycles were 3; the freezing time was 0.5 h and thawing temperature was 40℃. Both of the precision of the two methods were good.
The characteristics and qualities of both MCFAs nanoliposomesⅠandⅡwere evaluated. Compared with MCFAs nanoliposomesⅡ, MCFAs nanoliposomesⅠexhibited great advantages in their characteristics with regular spherical or elliptical in shape, a small average diameter (76.2±1.2 nm), narrow size distribution (PDI=0.207±0.003), high zeta potential (-50.51±0.4 mV), large entrapment efficiency (70.5±6.5%) and drug loading (9.4±0.4%), low surface tension (33.86±0.36 dyne/cm), high Tm (138℃), low leakage rate and good long-time storage stability.
The easy-energy-supply property of these two kinds of MCFAs nanoliposomes were estimated by the relive-fatigue experiments to the mice. The high-dose groups ofⅠandⅡ(680 mg/kg) suggested a significantly longer weight-loaded swimming time (104±29 min and 105±31min, respectively) and a higher hepatic glycogen (HG) (16.40±1.45 mg/g and 14.98±3.56 mg/g, respectively) than the control group. Moreover, the serum urea nitrogen (SUN) (891.5±113.4mg/L and 839.5±111.9mg/L, respectively) and blood lactic acid (LD) (6.05±1.40mmol/L and 5.70±1.02mmol/L, respectively) of high-dose groups were also significantly lower than those of the control group. However, the MCFAs group and the two MCFAs nanoliposomes groups showed no significant difference.
The suppression accumulation of body fat property of MCFAs nanoliposomesⅠandⅡwas investigated by short-term (two weeks) and long-term (five weeks) oral administration to the mice. Compared to the control group, the body weight of the mice, food intake, the adipose tissue and liver weights of the two MCFAs nanoliposomes were no significant difference. Nevertheless, the triglyceride (0.550±0.140 mmol/L and 0.384±0.150 mmol/L, respectively) and total cholesterol (0.621±0.097 and 0.540±0.247, respectively) were significantly lower than the control group. The MCFAs group and the tow MCFAs nanoliposomes groups showed no significant difference.
All the results indicated that MCFAs nanoliposomes prepared by thin-layer dispersion-DHPM and freeze-thawing-DHPM methods have overcome the drawbacks of MCFAs and exhibited good physicochemical properties. The easy-energy-supply and suppression of body fat accumulation characteristics revealed the MCFAs nanoliposomes can be a potential easy-energy-supply drug candidate.
引文
[1]Bach A C, Babayan V K. Medium-chain triglycerides:an update [J]. The American Journal of Clinical Nutrition,1982,36(5):950-962.
[2]Takeuchi H, Sekine S, Seto A. Medium-chain fatty acids-Nutritional function and application to cooking oil [J]. Lipid Technology,2008,20(1):9-12.
[3]Temme E H, Mensink R P, Hornstra G. Effects of medium chain fatty acids (MCFA), myristic acid, and oleic acid on serum lipoproteins in healthy subjects [J]. Journal of lipid research,1997, 38(9):1746-1754.
[4]Babayan V K. Medium chain triglycerides and structured lipids [J]. Lipids,1987,22(6): 417-420.
[5]Tsuji H, Kasai M, Takeuchi H, et al. Dietary medium-chain triacylglycerols suppress accumulation of body fat in a double-blind, controlled trial in healthy men and women [J]. Human Nutrition and Metabolism,2001,131(11):2853-2859.
[6]Jeukendrup A E, Aldred S. Fat supplementation, health, and endurance performance [J]. Nutrition,2004,20(7-8):678-688.
[7]Decuypere J A, Dierick N A. The combined use of triacylglycerols containing medium-chain fatty acids and exogenous lipolytic enzymes as an alternative to in-feed antibiotics in piglets: concept, possibilities and limitations. Nutrition Research Reviews. An overview,2003,16(2): 193-210.
[8]Traul K A, Driedger A, Ingle D L, et al. Review of the toxicologic properties of medium-chain triglycerides [J]. Food and Chemical Toxicology,2000,38(1):79-98.
[9]Traul K A, Driedger A, Ingle D L, et al. Review of the toxicologic properties of medium-chain triglycerides [J]. Food and Chemical Toxicology,2000,38(1):79-98.
[10]Steinbuch, M. and Audran, R. The isolation of IgG from mammalian sera with the aid of caprylic acid [J]. Archives of Biochemistry and Biophysics,1969,134:279-284.
[11]Rukmini C, Raghuram C C. Nutritional and biochemical aspects of the hypolipidemic action of rice bran:a review [J]. Journal of the American College of Nutrition,1991,10:593-601.
[12]Megremis C J. Medium chain triglycerides:a non-conventional fat [J]. Food Technology, 1991,45,108-110.
[13]Aoyama T, Nosaka N, Kasai M. Research on the nutritional characteristics of medium-chain fatty acids [J]. Journal of Medical Investigation.2007,54(3-4):385-388.
[14]Arifin N, Cheong L Z, Koh S P, et al. Physicochemical Properties and Sensory Attributes of Medium-and Long-Chain Triacylglycerols (MLCT)-Enriched Bakery Shortening [J]. Food and Bioprocess Technology.2009. DOI:10.1007/s1 1947-009-0204-0.
[15]St Onge M P, Ross R, Parsons W D, et al. Medium-chain triglycerides increase energy expenditure and decrease adiposity in overweight men [J]. Obesity Research,2003,11:395-402.
[16]Kasai M, Maiki H, Nosaka N, et al. Effect of medium-chain triglycerides on the postprandial triglyceride concentration in healthy men [J]. Bioscience, Biotechnology, and Biochemistry.2003, 67(1):46-53.
[17]Krothkiewski M. Value of VLCD supplementation with medium chain triglycerides [J]. International Journal of Obesity.2001,25(9):1393-1400.
08] Bangham A D, Standish M M, Watkins J C. Diffusion of univalentions across thelamellae of swollen phospholipids [J]. Journal of Molecular Biology,1965,13:238-252.
[19]Mulvey M, Huntgren S. Bacterial spelunkers [J]. Science,2000,289:732-733.
[20]Neville M, Richau K, Boni L, et al. A comparison of biodistribution of liposomal and soluble IL-2 by a new method based on time-resolved fluorometry of europium [J]. Cytokine,2000, 12(11):1702-1711.
[21]Lonez C, Vandenbranden M, Ruysschaert J M. Cationic liposomal lipids:from gene carriers to cell signaling [J]. Progress in Lipid Research,2008,47(5):340-347.
[22]Klibanov A L, Huang L. Long-circulating liposomes:development and perspectives [J]. Journal of liposome research,1992,2(3):321-334.
[23]Torchilin V P. Recent advances with liposomes as pharmaceutical carriers [J]. Nature Reviews Drug Discovery,2005,4(2):145-60.
[24]Bangham A D, Standish M M, Watkins J C. The action of steroids and steptolysin S on the permeability of phospholipid structures to cations [J]. Journal of Molecular Biology,1965, 13:138.
[25]Barenholz Y, Gibbes D, Litman B J, et al. A simple method for the preparation of homogeneous phospholipid vesicles [J]. Biochemistry.1977,16(12):2806-0810.
[26]Pick U. Liposomes with a large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures [J]. Archive Biochemistry and Biophysics,1981,212(1): 186-194.
[27]Szoka F, Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse phase evaporation [J]. Biochemistry,1978,75(9): 4194-4198.
[28]Barnadas-Rodriguez R, Sabes M. Factors involved in the production of liposomes with a high-pressure homogenizer [J]. International Journal of Pharmaceutics,2001,213(1-2):175-186.
[29]Zheng S, Alkan-Onyuksel H, Beissinger RL, et al. Liposome microencapsulations without using any organic solvent [J]. Journal of Dispersion Science and Technology,1999,20(4): 1189-1203.
[30]陈丽凤,张西臣,杨永胜,等.脂质体的制备及其在兽医学中的应用[J].中国兽医寄生虫病.1999,7(4)：57-61.
[31]Mozafari MR, Khosravi-Darani K, Borazan G G, et al. Encapsulation of food ingredients using nanoliposome technology [J]. International Journal of Food properties,2008,11:833-844.
[32]Taylor TM, Davidson PM, Bruce BD, et al. Liposomal nanocapsules in food science and agriculture [J]. Critical Reviews in Food Science and Nutrition,2005,45(7-8):587-605.
[33]Reineccius G A. Flavour manufacturing [M]. New Youk:In source bood of flavours; Reineccius GA; Chapman and Hall,1994,572-576.
[34]Paul P. Technological Properties of High Pressure Homogenizers:The Effect of Fat Globules, Milk Proteins, and Polysaccharides [J]. International Dairy Journal,1999,9:329-335.
[35]Liu W, Liu J H, Xie M Y, et al. Characterization and High Pressure icrofluidization-induced Activation of Polyphenoloxidase from Chinese Pear (Pyrus Pyrifolia Nakai) [J]. Journal of Agricultural and Food Chemistry,2009,57 (12):5376-5380.
[36]Kim H Y; Baianu I C. Novel liposome microencapsulation techniques for food applications [J]. Trends Food Science Technology,1991,2:55-61.
[37]Pupo E, Padron A, Santana E, et al. Preparation of plasmid DNA-containing liposomes using a high-pressure homogenization-extrusion technique [J]. Journal of Controlled Release,2005, 105:379-396.
[38]Zheng S, Alkan-Onyusel H, Beissinger R L, et al. Liposome meicroencapsulations without using any organic solvent [J]. Journal of dispersion science and technology,1999,20(4): 1189-1203.
[39]Vuillemard J C. Recent advances in the large-scale production of lipid vesicles for use in food products:microfluidization [J]. Journal of microencapsulation,1991,8(4):547-562.
[40]Kasaai M R, Charlet G, Paquin P, et al. Fragmentation of chitosan by microfluidization process [J]. Innovative Food Science Emerging Technology,2003,4(4):403-413.
[41]Maa Y F, Hsu C C. Performance of sonoication and microfluidization for liquid-liquid emulsification [J]. Pharm. Dev. Technol.1999,4:233-240.
[42]Mayhew E, Lazo R, Vail W J, et al. Characterization of liposomes prepared using a microemulsifier [J].Biochimica et Biophysica Acta,1984,775:169-174.
[43]Washington C, Davis S S, The production of parenteral feeding emulsions by Microfluidizer [J]. International Journal of Pharmaceutics,1988,44:169-176.
[44]Gregoriadis G S H, Florence A T. A procedure for the efficient entrapment of drugs in dehydration rehydration liposomes (DRVs) [J]. International Journal of Pharmaceutics,1990,65: 235-242.
[45]Barenholz. Y, Liposome production:historical aspects [M]. New York:Liposome Dermatics, Springer-Verlag,1992,69-81.
[46]Barnadas-Rodriguez R, Sabes M. Factors involved in the production of liposomes with a high-pressure homogenizer [J]. International Journal of Pharmaceutics,2001,213:175-186.
[47]Takahashi M, Kitamoto D, Imura T, et al. Characterization and bioavailability of liposomes containing a ukon extract [J]. Bioscience, Biotechnology, and Biochemistry,2008,72(5): 1199-1205.
[48]潘家祯,崔宁.用超高压微射流技术制备纳米级材料[C].纳微粉体制备与技术应用研讨会.北京,2002.
[49]张小宁,张郁,姬海红,等.微射流法制备莪术油纳米脂质体的研究[J].中国药学杂志 2004,39(5)：356-358.
[50]于波涛.提高脂质体包封率的方法及其研究进展[J].中国医药工业杂志,2002,2300：564-568
[51]冯怡,沈岚,徐德生,等.麦冬皂苷肠溶微球的载药量和包封率研究[J].中成药.2004,26(8)：611-613.
[52]Mozafari M R. Liposomes:an overview of manufacturing techniques [J]. Cellular and Molecular Biology Letters,2005,10:711-719.
[53]Picon A, Gaya P, Medina M, et al. The Effect of Liposome Encapsulation of Chymosin Derived by Fermentation on Manchego Cheese Ripening [J]. Journal of Dairy Science,1994,77: 16-23.
[54]Lasic D D, Joannic R, Keller B C, et al. Spontaneous Vesiculation [J]. Advances in Colloid and Interface Science,2001,89-90:337-349.
[55]Vemuri S, Rhodes C T. Preparation and Characterization of Liposomes as Therapeutic Delivery Systems:A Review [J]. Pharmaceutica Acta Helvetiae,1995,70:95-111.
[56]Coderch L, Fonollosa J, De Pera M, et al. Influence of cholesterol on liposome fluidity by EPR. Relationship with percutaneous absorption [J]. Journal of controlled release,2000,68(1): 85-95.
[57]Sulkowski W W, Pentak D, Nowak K. et al. The influence of temperature, cholesterol content and pH on liposome stability [J]. Journal of Molecular Structure,2005,744-747:737-747.
[58]Carrion C, Domingo J C, Madariaga M A. Preparation of long-circulating immuno-liposomes using PEG-cholesterol conjugates:effect of the spacer arm between PEG and cholesterol on liposomal characteristics [J]. Chemistry and Physics of Lipids,2001,113:97-110.
[59]Tasi L M, Liu D Z, Chen W Y. Microcalorimetric investigation of the interaction of polysorbate surfactants with unilamellar phosphatidylcholines liposomes [J]. Collids and surfaces A,2003,213:7-14.
[60]Xia S Q, Xu S Y. Ferrous sulfate liposomes:preparation, stability and application in fluid milk [J]. Food Research International,2005,38:289-296.
[61]Brandl M, Bachmann D, Drechsler M, et al. Liposome Preparation by a New High Pressure Homogenizer Gaulin Micron Lab 40 [J]. Drug Development and Industrial Phannacy,1990, 16(14):2167-2191.
[62]Barnadas-Rodriguez R, Sabes M. Factors Involved in the Production of Liposomes with a High-pressure Homogenizer [J]. International Journal of Pharmaceutics,2001. 213:175-186.
[63]Takahashi M, Inafuku K, Miyagi T, et al. Efficient Preparation of Liposomes Encapsulating Food Materials Using Lecithins by a Mechanochemical Method [J]. Journal of Oles Science,2007, 56(1):35-42.
[64]Traikia M, Warschawski D E, Recouvreur M, et al. Formation of unilamellar vesicles by repetitive freeze-thaw cycles:characterization by electron microscopy and 31P-nuclear magnetic resonance [J]. European Biophysics Journal,2000,29(3):184-195.
[65]Mayer L D, Hope M J, Cullis P R, et al. Solute distributions and trapping efficiencies observed in freeze-thawed multilamellar vesicles. Biochim Biophys Acta,1985,817:193-196.
[66]Miyajima. Koichiro Role of saccharides for the freeze-thawing and freeze drying of liposome [J]. Advanced Drug Delivery Reviews,1997,24(2,3):151-159.
[67]Pick U L. Liposomes with a large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures [J]. Archives of Biochemistry Biophysics,1981:212(1): 1986-194.
[68]Sriwongsitanont S, Ueno M. Effect of freeze-thawing and polyethylene glycon (PEG) lipid on fusion and fission of phospholipid vesicles [J]. Chemical & Pharmaceutical Bulletin,2004, 52(5):641-642.
[69]Ueno M, Sriwongsitanont S. Effect of PEG lipid on fusion and fission of phospholipid vesicles in the process of freeze-thawing [J]. Polymer.2005,46:1257-1267.
[70]张灵芝.脂质体的制备及其在生物医学中的应用[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.
[71]Kajiwara E, Kawano K, Hattori Y, et al. Long-circulating liposome-encapsulated ganciclovir enhances the efficacy of HSV-TK suicide gene therapy [J]. Journal of Control Release,2007, 120(1-2):104-110.
[72]Koopman J, Turkish V, Monto A. Infant formula and gastrointestinal illness [J]. American Journal of Public Health,1985,75:477-480.
[73]Crook T, Tinklenberg J, Yesavage J, et al. Effects of phosphatidylserine in age-associated memory impairment [J]. Neurology,1991,41:644-649.
[74]Yuh-Fun M, Chung C H. Performance of sonication and microfluidization for liquid-liquid emulsification [J]. Pharmaceutical Development and Technology,1999,4(2):233-240.
[75]Gruner S M, Lenk R P, Janoff A S, et al. Novel multilayered lipid vesicles:comparison of physical characteristics of multilamellar liposomes and stable plurilamellar vesicles [J]. Biochemistry,1985,24(4):2833-2842.
[76]Heurtault B, Saulnier P, Pech B, et al. Physico-chemical stability of colloidal lipid particles [J]. Biomaterials,2003,24:4283-4300.
[77]DeLuca T, Kaszuba M & Mattison K. Optimizing silicone emulsion stability using zeta potential [J]. American Laboratory,2006,38:14-15.
[78]Cui J X, Li C L, Guo W M, et al. Direct comparison of two pegylated liposomal doxorubicin formulations:Is AUC predictive for toxicity and efficacy? [J] Journal of Controlled Release,2007, 118:204-215.
[79]Liu N, Park H J. Chitosan-coated nanoliposome as vitamin E carrier [J]. Journal of Microencapsulation,2009,26:235-242.
[80]Labbe C, Crowe L M, Crowe J H. Stability of the lipid component of trout sperm plasma membrane during freeze-thawing [J]. Cryobiology,1997,34:176-182.
[81]Demetzos C. Differential scanning calorimetry (DSC):a tool to study the thermal behavior of lipid bilayers and liposomal stability [J]. Journal of Liposome Research,2008,18:159-173.
[82]Poste G. Liposome targeting in vivo:problems and opportunities [J]. Biology of the Cell, 1983,47:19-37.
[83]范明辉.红景天苷纳米脂质体的研制[D],江南大学,2008.
[84]Law L, Huang K J, Chou H Y. Preparation of desmopressin-containing liposomes for intranasal delivery [J]. Journal of Controlled Release,2001,70(3):375-382.
[85]Wolfram G. Medium-chain triglycerides (MCT) for total parenteral nutrition [J]. World Journal of Surgery,1986,10:33-37.
[86]Guillot E, Vaugelade P, Lemarchali P, et al. Intestinal absorption and liver uptake of medium-chain fatty acids in non-anaesthetized pigs [J]. British Journal of Nutrition,1993,69: 431-442.
[87]Megremis C J. Medium-chain triglycerides:a nonconventional fat [J]. Food Technology, 1991,45:108-110.
[88]Decuypere J A, Dierick N A. The combined use of triacylglycerols containing medium-chain fatty acids and exogenous lipolytic enzymes as an alternative to in-feed antibiotics in piglets: concept, possibilities and limitations [J]. An overview. Nutrition Research Reviews,2003,16: 193-210.
[89]Chen Y, Kong L D, Xia X, et al. Behavioral and biochemical studies of total furocoumarins from seeds of Psoralea corylifolia in the forced swimming test in mice [J]. Journal of Ethnopharmacology,2005,96(3):451-459.
[90]Hui Y H, Huang N H, Ebbert L, et al. Pharmacokinetic comparisons of tail-bleeding with cannula-or retro-orbital bleeding techniques in rats using six marketed drugs [J]. Journal of Pharmacological Toxicological Methods,2007,56:256-64.
[91]Belluardo N, Westerblad H, Mudo G, et al. Neuromuscular junction disassembly and muscle fatigue in mice lacking neurotrophin-4 [J]. Molecular and Cell Neuroscience,2001,18:56-67.
[92]Mague S D, Pliakas A M, Todtenkopf M S, et al. Antidepressant-like effects of κ-opioid receptor antagonists in the forced swim test in rats [J]. Journal of Pharmacology and Experimental Therapeutics,2003,305:323-330.
[93]Lapvetelainen T, Tiihonen A, Koskela P, et al. Training a large number of laboratory mice using running wheels and analyzing running behavior by use of a computer-assisted system [J]. Laboratory Animal Science,1997,47:172-179.
[94]Siekmeier R, Scheuch G. Systemic treatment by inhalation of macromolecules-principles, problems, and examples [J]. Journal of Physiology and Pharmacology,2008,59:53-79.
[95]Hettema E H, Tabak H F. Transport of fatty acids and metabolites across the peroxisomal membrane [J]. Biochim Biophys Acta,2000,1486:18-27.
[96]Tang W Y, hang Y Z, Gao J, et al. The anti-fatigue effect of 20 (R)-Ginsenoside Rg3 in mice by intranasally administration [J]. Biological and Pharmaceutical Bulletin,2008,31:2024-2027.
[97]Zhang Y, Yao X B, Bao B L, et al. Anti-fatigue activity of a triterpenoid-rich extract from Chinese bamboo shavings (Caulis bamfusae in taeniam) [J]. Phytotherapy Research,2006,20: 872-6.
[98]Tsopanakis C, Tsopanakis A. Stress hormonal factors, fatigue, and antioxidant responses to prolonged speed driving [J]. Pharmacology Biochemistry and Behavior,1998,60:747-751.
[99]An H J, Choi H M, Park H S, et al. Oral administration of hot water extracts of Chlorella vulgaris increases physical stamina in mice [J]. Annals of Nutrition and Metabolism,2006,50: 380-386.
[100]Tamminga S. The effect of the supply of rumen degradable protein and metabolisable protein on negative energy balance and fertility in dairy cows [J]. Animal Reproduction Science,2006,96: 227-239.
[101]Sahlin K, Tonkonogi M, Soderlund K. Energy supply and muscle fatigue in humans [J]. Acta Physiologica Scandinavica,1998,162:261-6.
[102]Okazaki Y, Ito Y, Kyo K, et al. Corrosion resistance and corrosion fatigue strength of new titanium alloys for medical implants without V and AI [J]. Material Science and Engineering:A, 1996,213:138-147
[103]Ma L, Cai D L, Li H X, et al. Anti-fatigue effects of salidroside in mice [J]. Journal of Medical Colleges of PLA,2008,23:88-93.
[104]Young A J, Castellani J W. Exertion-induced fatigue and thermoregulation in the cold [J]. Comparative Biochemistry Physiology Part A:Molecular and Integrative Physiology,2001,128: 759-776
[105]Bergstrom J, Hultman E. A study of the glycogen metabolism during exercise in man [J]. Scandinavian Journal of Clinical and Laboratory Investigation,1967,19:218-228.
[106]Azevedo Jr J L, Linderman J K, Lehman SL, et al. Training decreases muscle glycogen turnover during exercise [J]. European Journal of Applied Physiology,1998,78:479-486.
[107]Birkhahn R H, Border J R. Alternate or supplemental energy sources [J]. Journal of Parenteral and Enteral Nutrition,1981,5:24-31.
[108]Seaton TB, Welle S, Warenko M, et al. Thermic e□ect of medium-chain and long-chain triglycerides in man [J]. American Journal of Clinical Nutrition.1986,44 (5):630-634.
[109]Greenberger N J, Skillman T G. Medium-chain triglycerides [J]. The New England Journal of Medicine,1969,280:1045-1058.
[110]Nosaka N, Maki H, Suzuki Y, et al. E□ects of margarine containing medium-chain triacylglycerols on body fat reduction in humans [J]. Journal of Atherosclerosis and Thrombosis, 2003,10 (5),290-298.
[111]Matsuo T, Takeuchi H. E□ects of structured medium-and long-chain triacylglycerols in diets with various levels of fat on body fat accumulation in rats [J]. The British Journal of Nutrition, 2004,91:219-225.
[112]Matulka R A, Noguchi O, Nosaka N. Safety evaluation of a medium-and long-chain triacylglycerol oil produced from medium-chain triacylglycerols and edible vegetable oil [J]. Food and Chemical Toxicology.2006,44:1530-1538.
[113]Bray G A, Lee M, Bray T L. Weight-gain of rats fed medium-chain triglycerides is less than rats fed long-chain triglycerides [J]. International Journal of Obesity,1980,4:27-32.
[114]Noguchi O, Takeuchi H, Kubota F, et al. Larger diet-induced thermogenesis and less body fat accumulation in rats fed medium-chain triacylglycerols than in those fed long-chain triacylglycerols [J]. Journal of Nutritional Science and Vitaminology,2002,48:524-529.
[115]Papamandjaris A A, MacDougall D E, Jones P J H. Medium chain fatty acid metabolism and energy expenditure:Obesity treatment implications [J]. Life Sciences,1998,62:1203-1215.
[116]Hill J O, Peters J C, Swift L L, et al. Changes in blood-lipids during 6 days of overfeeding with medium or long-chain triglycerides [J]. Journal of Lipid Research,1990,31:407-416.
[117]Shinohara H, Shimada H, Noguchi O, et al. Effect of medium-chain fatty acids-containing dietary oil on hepatic fatty acid oxidation enzyme activity in rats [J]. Journal of Oleo Science, 2002,51(10):621-626.
[118]Takeuchi H, Sekine S, Seto A. Medium-chain fatty acids-nutritional function and application to cooking oil [J]. European Journal of Lipid Science andTechnology,2008,20(1):9-12.
019] Guo W, Choi J K, Kirkland J L, et al. Esterification of free fatty acids in adipocytes:a comparison between octanoate and oleate [J]. Biochemical Journal,2000,349:463-471.
[120]Wahlqvist M L. Dietary fat and the prevention of chronic disease [J]. Asia Pacific Journal of Clinical Nutrition,2005,14(4):313-318.
[121]Ikeda I, T omari Y, Sugano M, et al. Lymphatic Absorption of Structured Glycerolipids containing medium-chain fatty acids and linoleic acid, and their effect on cholesterol absorption in rats [J]. Lipids,1991,26(5):369-373.
[2]Takeuchi H, Sekine S, Seto A. Medium-chain fatty acids-Nutritional function and application to cooking oil [J]. Lipid Technology,2008,20(1):9-12.
[3]Temme E H, Mensink R P, Hornstra G. Effects of medium chain fatty acids (MCFA), myristic acid, and oleic acid on serum lipoproteins in healthy subjects [J]. Journal of lipid research,1997, 38(9):1746-1754.
[4]Babayan V K. Medium chain triglycerides and structured lipids [J]. Lipids,1987,22(6): 417-420.
[5]Tsuji H, Kasai M, Takeuchi H, et al. Dietary medium-chain triacylglycerols suppress accumulation of body fat in a double-blind, controlled trial in healthy men and women [J]. Human Nutrition and Metabolism,2001,131(11):2853-2859.
[6]Jeukendrup A E, Aldred S. Fat supplementation, health, and endurance performance [J]. Nutrition,2004,20(7-8):678-688.
[7]Decuypere J A, Dierick N A. The combined use of triacylglycerols containing medium-chain fatty acids and exogenous lipolytic enzymes as an alternative to in-feed antibiotics in piglets: concept, possibilities and limitations. Nutrition Research Reviews. An overview,2003,16(2): 193-210.
[8]Traul K A, Driedger A, Ingle D L, et al. Review of the toxicologic properties of medium-chain triglycerides [J]. Food and Chemical Toxicology,2000,38(1):79-98.
[9]Traul K A, Driedger A, Ingle D L, et al. Review of the toxicologic properties of medium-chain triglycerides [J]. Food and Chemical Toxicology,2000,38(1):79-98.
[10]Steinbuch, M. and Audran, R. The isolation of IgG from mammalian sera with the aid of caprylic acid [J]. Archives of Biochemistry and Biophysics,1969,134:279-284.
[11]Rukmini C, Raghuram C C. Nutritional and biochemical aspects of the hypolipidemic action of rice bran:a review [J]. Journal of the American College of Nutrition,1991,10:593-601.
[12]Megremis C J. Medium chain triglycerides:a non-conventional fat [J]. Food Technology, 1991,45,108-110.
[13]Aoyama T, Nosaka N, Kasai M. Research on the nutritional characteristics of medium-chain fatty acids [J]. Journal of Medical Investigation.2007,54(3-4):385-388.
[14]Arifin N, Cheong L Z, Koh S P, et al. Physicochemical Properties and Sensory Attributes of Medium-and Long-Chain Triacylglycerols (MLCT)-Enriched Bakery Shortening [J]. Food and Bioprocess Technology.2009. DOI:10.1007/s1 1947-009-0204-0.
[15]St Onge M P, Ross R, Parsons W D, et al. Medium-chain triglycerides increase energy expenditure and decrease adiposity in overweight men [J]. Obesity Research,2003,11:395-402.
[16]Kasai M, Maiki H, Nosaka N, et al. Effect of medium-chain triglycerides on the postprandial triglyceride concentration in healthy men [J]. Bioscience, Biotechnology, and Biochemistry.2003, 67(1):46-53.
[17]Krothkiewski M. Value of VLCD supplementation with medium chain triglycerides [J]. International Journal of Obesity.2001,25(9):1393-1400.
08] Bangham A D, Standish M M, Watkins J C. Diffusion of univalentions across thelamellae of swollen phospholipids [J]. Journal of Molecular Biology,1965,13:238-252.
[19]Mulvey M, Huntgren S. Bacterial spelunkers [J]. Science,2000,289:732-733.
[20]Neville M, Richau K, Boni L, et al. A comparison of biodistribution of liposomal and soluble IL-2 by a new method based on time-resolved fluorometry of europium [J]. Cytokine,2000, 12(11):1702-1711.
[21]Lonez C, Vandenbranden M, Ruysschaert J M. Cationic liposomal lipids:from gene carriers to cell signaling [J]. Progress in Lipid Research,2008,47(5):340-347.
[22]Klibanov A L, Huang L. Long-circulating liposomes:development and perspectives [J]. Journal of liposome research,1992,2(3):321-334.
[23]Torchilin V P. Recent advances with liposomes as pharmaceutical carriers [J]. Nature Reviews Drug Discovery,2005,4(2):145-60.
[24]Bangham A D, Standish M M, Watkins J C. The action of steroids and steptolysin S on the permeability of phospholipid structures to cations [J]. Journal of Molecular Biology,1965, 13:138.
[25]Barenholz Y, Gibbes D, Litman B J, et al. A simple method for the preparation of homogeneous phospholipid vesicles [J]. Biochemistry.1977,16(12):2806-0810.
[26]Pick U. Liposomes with a large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures [J]. Archive Biochemistry and Biophysics,1981,212(1): 186-194.
[27]Szoka F, Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse phase evaporation [J]. Biochemistry,1978,75(9): 4194-4198.
[28]Barnadas-Rodriguez R, Sabes M. Factors involved in the production of liposomes with a high-pressure homogenizer [J]. International Journal of Pharmaceutics,2001,213(1-2):175-186.
[29]Zheng S, Alkan-Onyuksel H, Beissinger RL, et al. Liposome microencapsulations without using any organic solvent [J]. Journal of Dispersion Science and Technology,1999,20(4): 1189-1203.
[30]陈丽凤,张西臣,杨永胜,等.脂质体的制备及其在兽医学中的应用[J].中国兽医寄生虫病.1999,7(4)：57-61.
[31]Mozafari MR, Khosravi-Darani K, Borazan G G, et al. Encapsulation of food ingredients using nanoliposome technology [J]. International Journal of Food properties,2008,11:833-844.
[32]Taylor TM, Davidson PM, Bruce BD, et al. Liposomal nanocapsules in food science and agriculture [J]. Critical Reviews in Food Science and Nutrition,2005,45(7-8):587-605.
[33]Reineccius G A. Flavour manufacturing [M]. New Youk:In source bood of flavours; Reineccius GA; Chapman and Hall,1994,572-576.
[34]Paul P. Technological Properties of High Pressure Homogenizers:The Effect of Fat Globules, Milk Proteins, and Polysaccharides [J]. International Dairy Journal,1999,9:329-335.
[35]Liu W, Liu J H, Xie M Y, et al. Characterization and High Pressure icrofluidization-induced Activation of Polyphenoloxidase from Chinese Pear (Pyrus Pyrifolia Nakai) [J]. Journal of Agricultural and Food Chemistry,2009,57 (12):5376-5380.
[36]Kim H Y; Baianu I C. Novel liposome microencapsulation techniques for food applications [J]. Trends Food Science Technology,1991,2:55-61.
[37]Pupo E, Padron A, Santana E, et al. Preparation of plasmid DNA-containing liposomes using a high-pressure homogenization-extrusion technique [J]. Journal of Controlled Release,2005, 105:379-396.
[38]Zheng S, Alkan-Onyusel H, Beissinger R L, et al. Liposome meicroencapsulations without using any organic solvent [J]. Journal of dispersion science and technology,1999,20(4): 1189-1203.
[39]Vuillemard J C. Recent advances in the large-scale production of lipid vesicles for use in food products:microfluidization [J]. Journal of microencapsulation,1991,8(4):547-562.
[40]Kasaai M R, Charlet G, Paquin P, et al. Fragmentation of chitosan by microfluidization process [J]. Innovative Food Science Emerging Technology,2003,4(4):403-413.
[41]Maa Y F, Hsu C C. Performance of sonoication and microfluidization for liquid-liquid emulsification [J]. Pharm. Dev. Technol.1999,4:233-240.
[42]Mayhew E, Lazo R, Vail W J, et al. Characterization of liposomes prepared using a microemulsifier [J].Biochimica et Biophysica Acta,1984,775:169-174.
[43]Washington C, Davis S S, The production of parenteral feeding emulsions by Microfluidizer [J]. International Journal of Pharmaceutics,1988,44:169-176.
[44]Gregoriadis G S H, Florence A T. A procedure for the efficient entrapment of drugs in dehydration rehydration liposomes (DRVs) [J]. International Journal of Pharmaceutics,1990,65: 235-242.
[45]Barenholz. Y, Liposome production:historical aspects [M]. New York:Liposome Dermatics, Springer-Verlag,1992,69-81.
[46]Barnadas-Rodriguez R, Sabes M. Factors involved in the production of liposomes with a high-pressure homogenizer [J]. International Journal of Pharmaceutics,2001,213:175-186.
[47]Takahashi M, Kitamoto D, Imura T, et al. Characterization and bioavailability of liposomes containing a ukon extract [J]. Bioscience, Biotechnology, and Biochemistry,2008,72(5): 1199-1205.
[48]潘家祯,崔宁.用超高压微射流技术制备纳米级材料[C].纳微粉体制备与技术应用研讨会.北京,2002.
[49]张小宁,张郁,姬海红,等.微射流法制备莪术油纳米脂质体的研究[J].中国药学杂志 2004,39(5)：356-358.
[50]于波涛.提高脂质体包封率的方法及其研究进展[J].中国医药工业杂志,2002,2300：564-568
[51]冯怡,沈岚,徐德生,等.麦冬皂苷肠溶微球的载药量和包封率研究[J].中成药.2004,26(8)：611-613.
[52]Mozafari M R. Liposomes:an overview of manufacturing techniques [J]. Cellular and Molecular Biology Letters,2005,10:711-719.
[53]Picon A, Gaya P, Medina M, et al. The Effect of Liposome Encapsulation of Chymosin Derived by Fermentation on Manchego Cheese Ripening [J]. Journal of Dairy Science,1994,77: 16-23.
[54]Lasic D D, Joannic R, Keller B C, et al. Spontaneous Vesiculation [J]. Advances in Colloid and Interface Science,2001,89-90:337-349.
[55]Vemuri S, Rhodes C T. Preparation and Characterization of Liposomes as Therapeutic Delivery Systems:A Review [J]. Pharmaceutica Acta Helvetiae,1995,70:95-111.
[56]Coderch L, Fonollosa J, De Pera M, et al. Influence of cholesterol on liposome fluidity by EPR. Relationship with percutaneous absorption [J]. Journal of controlled release,2000,68(1): 85-95.
[57]Sulkowski W W, Pentak D, Nowak K. et al. The influence of temperature, cholesterol content and pH on liposome stability [J]. Journal of Molecular Structure,2005,744-747:737-747.
[58]Carrion C, Domingo J C, Madariaga M A. Preparation of long-circulating immuno-liposomes using PEG-cholesterol conjugates:effect of the spacer arm between PEG and cholesterol on liposomal characteristics [J]. Chemistry and Physics of Lipids,2001,113:97-110.
[59]Tasi L M, Liu D Z, Chen W Y. Microcalorimetric investigation of the interaction of polysorbate surfactants with unilamellar phosphatidylcholines liposomes [J]. Collids and surfaces A,2003,213:7-14.
[60]Xia S Q, Xu S Y. Ferrous sulfate liposomes:preparation, stability and application in fluid milk [J]. Food Research International,2005,38:289-296.
[61]Brandl M, Bachmann D, Drechsler M, et al. Liposome Preparation by a New High Pressure Homogenizer Gaulin Micron Lab 40 [J]. Drug Development and Industrial Phannacy,1990, 16(14):2167-2191.
[62]Barnadas-Rodriguez R, Sabes M. Factors Involved in the Production of Liposomes with a High-pressure Homogenizer [J]. International Journal of Pharmaceutics,2001. 213:175-186.
[63]Takahashi M, Inafuku K, Miyagi T, et al. Efficient Preparation of Liposomes Encapsulating Food Materials Using Lecithins by a Mechanochemical Method [J]. Journal of Oles Science,2007, 56(1):35-42.
[64]Traikia M, Warschawski D E, Recouvreur M, et al. Formation of unilamellar vesicles by repetitive freeze-thaw cycles:characterization by electron microscopy and 31P-nuclear magnetic resonance [J]. European Biophysics Journal,2000,29(3):184-195.
[65]Mayer L D, Hope M J, Cullis P R, et al. Solute distributions and trapping efficiencies observed in freeze-thawed multilamellar vesicles. Biochim Biophys Acta,1985,817:193-196.
[66]Miyajima. Koichiro Role of saccharides for the freeze-thawing and freeze drying of liposome [J]. Advanced Drug Delivery Reviews,1997,24(2,3):151-159.
[67]Pick U L. Liposomes with a large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures [J]. Archives of Biochemistry Biophysics,1981:212(1): 1986-194.
[68]Sriwongsitanont S, Ueno M. Effect of freeze-thawing and polyethylene glycon (PEG) lipid on fusion and fission of phospholipid vesicles [J]. Chemical & Pharmaceutical Bulletin,2004, 52(5):641-642.
[69]Ueno M, Sriwongsitanont S. Effect of PEG lipid on fusion and fission of phospholipid vesicles in the process of freeze-thawing [J]. Polymer.2005,46:1257-1267.
[70]张灵芝.脂质体的制备及其在生物医学中的应用[M].北京：北京医科大学、中国协和医科大学联合出版社,1998.
[71]Kajiwara E, Kawano K, Hattori Y, et al. Long-circulating liposome-encapsulated ganciclovir enhances the efficacy of HSV-TK suicide gene therapy [J]. Journal of Control Release,2007, 120(1-2):104-110.
[72]Koopman J, Turkish V, Monto A. Infant formula and gastrointestinal illness [J]. American Journal of Public Health,1985,75:477-480.
[73]Crook T, Tinklenberg J, Yesavage J, et al. Effects of phosphatidylserine in age-associated memory impairment [J]. Neurology,1991,41:644-649.
[74]Yuh-Fun M, Chung C H. Performance of sonication and microfluidization for liquid-liquid emulsification [J]. Pharmaceutical Development and Technology,1999,4(2):233-240.
[75]Gruner S M, Lenk R P, Janoff A S, et al. Novel multilayered lipid vesicles:comparison of physical characteristics of multilamellar liposomes and stable plurilamellar vesicles [J]. Biochemistry,1985,24(4):2833-2842.
[76]Heurtault B, Saulnier P, Pech B, et al. Physico-chemical stability of colloidal lipid particles [J]. Biomaterials,2003,24:4283-4300.
[77]DeLuca T, Kaszuba M & Mattison K. Optimizing silicone emulsion stability using zeta potential [J]. American Laboratory,2006,38:14-15.
[78]Cui J X, Li C L, Guo W M, et al. Direct comparison of two pegylated liposomal doxorubicin formulations:Is AUC predictive for toxicity and efficacy? [J] Journal of Controlled Release,2007, 118:204-215.
[79]Liu N, Park H J. Chitosan-coated nanoliposome as vitamin E carrier [J]. Journal of Microencapsulation,2009,26:235-242.
[80]Labbe C, Crowe L M, Crowe J H. Stability of the lipid component of trout sperm plasma membrane during freeze-thawing [J]. Cryobiology,1997,34:176-182.
[81]Demetzos C. Differential scanning calorimetry (DSC):a tool to study the thermal behavior of lipid bilayers and liposomal stability [J]. Journal of Liposome Research,2008,18:159-173.
[82]Poste G. Liposome targeting in vivo:problems and opportunities [J]. Biology of the Cell, 1983,47:19-37.
[83]范明辉.红景天苷纳米脂质体的研制[D],江南大学,2008.
[84]Law L, Huang K J, Chou H Y. Preparation of desmopressin-containing liposomes for intranasal delivery [J]. Journal of Controlled Release,2001,70(3):375-382.
[85]Wolfram G. Medium-chain triglycerides (MCT) for total parenteral nutrition [J]. World Journal of Surgery,1986,10:33-37.
[86]Guillot E, Vaugelade P, Lemarchali P, et al. Intestinal absorption and liver uptake of medium-chain fatty acids in non-anaesthetized pigs [J]. British Journal of Nutrition,1993,69: 431-442.
[87]Megremis C J. Medium-chain triglycerides:a nonconventional fat [J]. Food Technology, 1991,45:108-110.
[88]Decuypere J A, Dierick N A. The combined use of triacylglycerols containing medium-chain fatty acids and exogenous lipolytic enzymes as an alternative to in-feed antibiotics in piglets: concept, possibilities and limitations [J]. An overview. Nutrition Research Reviews,2003,16: 193-210.
[89]Chen Y, Kong L D, Xia X, et al. Behavioral and biochemical studies of total furocoumarins from seeds of Psoralea corylifolia in the forced swimming test in mice [J]. Journal of Ethnopharmacology,2005,96(3):451-459.
[90]Hui Y H, Huang N H, Ebbert L, et al. Pharmacokinetic comparisons of tail-bleeding with cannula-or retro-orbital bleeding techniques in rats using six marketed drugs [J]. Journal of Pharmacological Toxicological Methods,2007,56:256-64.
[91]Belluardo N, Westerblad H, Mudo G, et al. Neuromuscular junction disassembly and muscle fatigue in mice lacking neurotrophin-4 [J]. Molecular and Cell Neuroscience,2001,18:56-67.
[92]Mague S D, Pliakas A M, Todtenkopf M S, et al. Antidepressant-like effects of κ-opioid receptor antagonists in the forced swim test in rats [J]. Journal of Pharmacology and Experimental Therapeutics,2003,305:323-330.
[93]Lapvetelainen T, Tiihonen A, Koskela P, et al. Training a large number of laboratory mice using running wheels and analyzing running behavior by use of a computer-assisted system [J]. Laboratory Animal Science,1997,47:172-179.
[94]Siekmeier R, Scheuch G. Systemic treatment by inhalation of macromolecules-principles, problems, and examples [J]. Journal of Physiology and Pharmacology,2008,59:53-79.
[95]Hettema E H, Tabak H F. Transport of fatty acids and metabolites across the peroxisomal membrane [J]. Biochim Biophys Acta,2000,1486:18-27.
[96]Tang W Y, hang Y Z, Gao J, et al. The anti-fatigue effect of 20 (R)-Ginsenoside Rg3 in mice by intranasally administration [J]. Biological and Pharmaceutical Bulletin,2008,31:2024-2027.
[97]Zhang Y, Yao X B, Bao B L, et al. Anti-fatigue activity of a triterpenoid-rich extract from Chinese bamboo shavings (Caulis bamfusae in taeniam) [J]. Phytotherapy Research,2006,20: 872-6.
[98]Tsopanakis C, Tsopanakis A. Stress hormonal factors, fatigue, and antioxidant responses to prolonged speed driving [J]. Pharmacology Biochemistry and Behavior,1998,60:747-751.
[99]An H J, Choi H M, Park H S, et al. Oral administration of hot water extracts of Chlorella vulgaris increases physical stamina in mice [J]. Annals of Nutrition and Metabolism,2006,50: 380-386.
[100]Tamminga S. The effect of the supply of rumen degradable protein and metabolisable protein on negative energy balance and fertility in dairy cows [J]. Animal Reproduction Science,2006,96: 227-239.
[101]Sahlin K, Tonkonogi M, Soderlund K. Energy supply and muscle fatigue in humans [J]. Acta Physiologica Scandinavica,1998,162:261-6.
[102]Okazaki Y, Ito Y, Kyo K, et al. Corrosion resistance and corrosion fatigue strength of new titanium alloys for medical implants without V and AI [J]. Material Science and Engineering:A, 1996,213:138-147
[103]Ma L, Cai D L, Li H X, et al. Anti-fatigue effects of salidroside in mice [J]. Journal of Medical Colleges of PLA,2008,23:88-93.
[104]Young A J, Castellani J W. Exertion-induced fatigue and thermoregulation in the cold [J]. Comparative Biochemistry Physiology Part A:Molecular and Integrative Physiology,2001,128: 759-776
[105]Bergstrom J, Hultman E. A study of the glycogen metabolism during exercise in man [J]. Scandinavian Journal of Clinical and Laboratory Investigation,1967,19:218-228.
[106]Azevedo Jr J L, Linderman J K, Lehman SL, et al. Training decreases muscle glycogen turnover during exercise [J]. European Journal of Applied Physiology,1998,78:479-486.
[107]Birkhahn R H, Border J R. Alternate or supplemental energy sources [J]. Journal of Parenteral and Enteral Nutrition,1981,5:24-31.
[108]Seaton TB, Welle S, Warenko M, et al. Thermic e□ect of medium-chain and long-chain triglycerides in man [J]. American Journal of Clinical Nutrition.1986,44 (5):630-634.
[109]Greenberger N J, Skillman T G. Medium-chain triglycerides [J]. The New England Journal of Medicine,1969,280:1045-1058.
[110]Nosaka N, Maki H, Suzuki Y, et al. E□ects of margarine containing medium-chain triacylglycerols on body fat reduction in humans [J]. Journal of Atherosclerosis and Thrombosis, 2003,10 (5),290-298.
[111]Matsuo T, Takeuchi H. E□ects of structured medium-and long-chain triacylglycerols in diets with various levels of fat on body fat accumulation in rats [J]. The British Journal of Nutrition, 2004,91:219-225.
[112]Matulka R A, Noguchi O, Nosaka N. Safety evaluation of a medium-and long-chain triacylglycerol oil produced from medium-chain triacylglycerols and edible vegetable oil [J]. Food and Chemical Toxicology.2006,44:1530-1538.
[113]Bray G A, Lee M, Bray T L. Weight-gain of rats fed medium-chain triglycerides is less than rats fed long-chain triglycerides [J]. International Journal of Obesity,1980,4:27-32.
[114]Noguchi O, Takeuchi H, Kubota F, et al. Larger diet-induced thermogenesis and less body fat accumulation in rats fed medium-chain triacylglycerols than in those fed long-chain triacylglycerols [J]. Journal of Nutritional Science and Vitaminology,2002,48:524-529.
[115]Papamandjaris A A, MacDougall D E, Jones P J H. Medium chain fatty acid metabolism and energy expenditure:Obesity treatment implications [J]. Life Sciences,1998,62:1203-1215.
[116]Hill J O, Peters J C, Swift L L, et al. Changes in blood-lipids during 6 days of overfeeding with medium or long-chain triglycerides [J]. Journal of Lipid Research,1990,31:407-416.
[117]Shinohara H, Shimada H, Noguchi O, et al. Effect of medium-chain fatty acids-containing dietary oil on hepatic fatty acid oxidation enzyme activity in rats [J]. Journal of Oleo Science, 2002,51(10):621-626.
[118]Takeuchi H, Sekine S, Seto A. Medium-chain fatty acids-nutritional function and application to cooking oil [J]. European Journal of Lipid Science andTechnology,2008,20(1):9-12.
019] Guo W, Choi J K, Kirkland J L, et al. Esterification of free fatty acids in adipocytes:a comparison between octanoate and oleate [J]. Biochemical Journal,2000,349:463-471.
[120]Wahlqvist M L. Dietary fat and the prevention of chronic disease [J]. Asia Pacific Journal of Clinical Nutrition,2005,14(4):313-318.
[121]Ikeda I, T omari Y, Sugano M, et al. Lymphatic Absorption of Structured Glycerolipids containing medium-chain fatty acids and linoleic acid, and their effect on cholesterol absorption in rats [J]. Lipids,1991,26(5):369-373.