神经生长因子脂质体的制备及药代动力学初步研究
摘要
目的
提纯神经生长因子(NGF)，并使用自制脂质体包裹，初步研究神经生长因子脂质体的药代动力学。
方法
上部为神经生长因子脂质体的制备。取小鼠颌下腺组织，经过离心、CM52上柱分离，得到NGF，电泳法检测纯度，Bradford法测定NGF含量，鸡胚背根神经节培养鉴定NGF活性。改进传统的反向蒸发法制备脂质体，并使用磁力搅拌法包裹神经生长因子，采用DEAE Sephadex A50微型柱离心法测定脂质体对NGF的包封率。
下部为静脉用NGF脂质体在家兔体内血药浓度检测及药代动力学参数的分析，并初步观察其组织分布，使用~(99m)Tc标记NGF，测定~(99m)Tc放射性计数与NGF含量的线性关系，从而设计检测家兔血浆中NGF浓度的检测方法，将家兔分为静脉注射单纯NGF组和脂质体包裹NGF组，通过TCA沉淀法处理血样，测定不同时间的血药浓度，使用3P97软件分析药代动力学参数及房室模型。使用SPECT检测NGF的全身分布，初步观察其代谢，搜集尿液分析代谢情况，另外使用ELISA法检测脑脊液中NGF，初步分析其在脑部分布。
结果
上部结果：提纯的NGF得率为125μg／g，通过电泳法显示提取的蛋白质为2.5S NGF，无明显杂质条带，鸡胚背根神经节培养显示活性良好，Bradford法测定NGF含量为1mg／ml。成功制备NGF脂质体静脉注射剂。
下部结果：通过同位素示踪法检测NGF含量与放射性计数成线
」il)洲卜人分一‘吹卜L片训t仑又
性关系，在剂量为40冬、g/kg时·脂质体包裹NGF的药代动力学参数
为:t，:。，一o.2338h，t，:j厂1 .322911，Al一;C一1 06.57(ng/l二I)*h，单纯NG「
的药代动力学参数为t.:《，一o.077h，t}:l于0.88h，AUC一98.656(ng/ml)*h，
SPECT显示脂质体包裹后的99mTc一NGF在脑部有浓聚，而未包裹则
无明显脑分布迹象，脑脊液ELISA检测显示只有脂质体NGF在用药
2，J、时左右检测到NGF，尿液及粪便放射性计数显示脂质体包裹NGF
为肾脏、胆汁排泄，而单纯NGF通过肾脏排泄，尿液沉淀实验显示
NGF肾脏排泄以小分子片段形式，而不是原型或大分子多肤片段形
式排出。
结论
自制脂质体包裹NGF后可以对NGF的吸收、分布、排泄、代谢
产生一定的变化，初步结果显示使用脂质体包裹后NGF代谢时间延
长并可能分布于脑组织。
Objective
To pack nerve growth factor with liposome and to study its pharmacokinetic parameters.
Methods
In first part, NGF-liposome was produced. Tissue of mice's submaxillary glands was centrifuged and separated in CM52 column, thus 2.5S NGF was produced. Electrophoresis was used to measure the purity and Bradford way to detect the content. To identify activity of NGF, cultivating DPG of chick embryo was performed. NGF was packed with liposome to produce NGF-liposome and correlative parameters were detected.
In second part, pharmacokinetics of NGF-liposome in vivo was studied. NGF was labeled with 99mTc and isotopic tracing was utilized to detect concentration of NGF in rabbit blood. Compared with packed group, unpacked NGF group were detected. Data were inputted into computer to be analyzed of pharmacokinetic parameters, compartment model etc. by software (3p97). From SPECT distribution of lipo-99mTc-NGF in rabbit body was shown as well 99mTc-NGF to be compared with. Urine and dejecta was collected for the analysis of excretion in rabbit body. ELISA was the method to detect concentration of NGF in CSF and TCA method was used to detect radioactivity of deposit of urine.
Results
In first part, it was indicated that the abstracted protein was NGF of which the content was ling/ml and biologic activity was fine. NGF was packed with liposome and NGF-liposome was produced.
In second part, we have found that the relationship of concentration of NGF and count of -ray was nearly linear. From the test of recovery and precision, it was suggested that labeling NGF with 99mTc to detect concentration of NGF was feasible. Pharmacokinetic parameters of NGF packed with liposome injected in vivo in rabbit body were: t1/2=0.2338h, t1/2=1.3229h, AUC=106.57(ng/ml)*h. Parameters of unpacked NGF were: t1/2 =0.077h, t1/2 =0.88h,AUC=98.656(ng/ml)*h. It was shown in pictures of SPECT, if NGF was packed with liposome there was NGF distributing in brain. Analysis of urine and dejecta indicated that main excretion route of unpacked NGF was urine and that of NGF packed with liposome was urine as well bile. Detection of NGF in CSF pointed that if NGF was packed peak of concentration in CSF was at 2hours whereas if NGF was unpacked NGF was undetectable. From SPECT, it was shown that there was radiological concentration in brain if 99mTc-NGF was packed with liposome. Radioactivity of urine deposit was u
ndetectable in TCA method.
Conclusion
When NGF is packed with liposome, absorption, distribution, metabolism and excretion of NGF will change. T1/2 and T1/2p are prolonged and NGF can pass BBB.
提纯神经生长因子(NGF)，并使用自制脂质体包裹，初步研究神经生长因子脂质体的药代动力学。
方法
上部为神经生长因子脂质体的制备。取小鼠颌下腺组织，经过离心、CM52上柱分离，得到NGF，电泳法检测纯度，Bradford法测定NGF含量，鸡胚背根神经节培养鉴定NGF活性。改进传统的反向蒸发法制备脂质体，并使用磁力搅拌法包裹神经生长因子，采用DEAE Sephadex A50微型柱离心法测定脂质体对NGF的包封率。
下部为静脉用NGF脂质体在家兔体内血药浓度检测及药代动力学参数的分析，并初步观察其组织分布，使用~(99m)Tc标记NGF，测定~(99m)Tc放射性计数与NGF含量的线性关系，从而设计检测家兔血浆中NGF浓度的检测方法，将家兔分为静脉注射单纯NGF组和脂质体包裹NGF组，通过TCA沉淀法处理血样，测定不同时间的血药浓度，使用3P97软件分析药代动力学参数及房室模型。使用SPECT检测NGF的全身分布，初步观察其代谢，搜集尿液分析代谢情况，另外使用ELISA法检测脑脊液中NGF，初步分析其在脑部分布。
结果
上部结果：提纯的NGF得率为125μg／g，通过电泳法显示提取的蛋白质为2.5S NGF，无明显杂质条带，鸡胚背根神经节培养显示活性良好，Bradford法测定NGF含量为1mg／ml。成功制备NGF脂质体静脉注射剂。
下部结果：通过同位素示踪法检测NGF含量与放射性计数成线
」il)洲卜人分一‘吹卜L片训t仑又
性关系，在剂量为40冬、g/kg时·脂质体包裹NGF的药代动力学参数
为:t，:。，一o.2338h，t，:j厂1 .322911，Al一;C一1 06.57(ng/l二I)*h，单纯NG「
的药代动力学参数为t.:《，一o.077h，t}:l于0.88h，AUC一98.656(ng/ml)*h，
SPECT显示脂质体包裹后的99mTc一NGF在脑部有浓聚，而未包裹则
无明显脑分布迹象，脑脊液ELISA检测显示只有脂质体NGF在用药
2，J、时左右检测到NGF，尿液及粪便放射性计数显示脂质体包裹NGF
为肾脏、胆汁排泄，而单纯NGF通过肾脏排泄，尿液沉淀实验显示
NGF肾脏排泄以小分子片段形式，而不是原型或大分子多肤片段形
式排出。
结论
自制脂质体包裹NGF后可以对NGF的吸收、分布、排泄、代谢
产生一定的变化，初步结果显示使用脂质体包裹后NGF代谢时间延
长并可能分布于脑组织。
Objective
To pack nerve growth factor with liposome and to study its pharmacokinetic parameters.
Methods
In first part, NGF-liposome was produced. Tissue of mice's submaxillary glands was centrifuged and separated in CM52 column, thus 2.5S NGF was produced. Electrophoresis was used to measure the purity and Bradford way to detect the content. To identify activity of NGF, cultivating DPG of chick embryo was performed. NGF was packed with liposome to produce NGF-liposome and correlative parameters were detected.
In second part, pharmacokinetics of NGF-liposome in vivo was studied. NGF was labeled with 99mTc and isotopic tracing was utilized to detect concentration of NGF in rabbit blood. Compared with packed group, unpacked NGF group were detected. Data were inputted into computer to be analyzed of pharmacokinetic parameters, compartment model etc. by software (3p97). From SPECT distribution of lipo-99mTc-NGF in rabbit body was shown as well 99mTc-NGF to be compared with. Urine and dejecta was collected for the analysis of excretion in rabbit body. ELISA was the method to detect concentration of NGF in CSF and TCA method was used to detect radioactivity of deposit of urine.
Results
In first part, it was indicated that the abstracted protein was NGF of which the content was ling/ml and biologic activity was fine. NGF was packed with liposome and NGF-liposome was produced.
In second part, we have found that the relationship of concentration of NGF and count of -ray was nearly linear. From the test of recovery and precision, it was suggested that labeling NGF with 99mTc to detect concentration of NGF was feasible. Pharmacokinetic parameters of NGF packed with liposome injected in vivo in rabbit body were: t1/2=0.2338h, t1/2=1.3229h, AUC=106.57(ng/ml)*h. Parameters of unpacked NGF were: t1/2 =0.077h, t1/2 =0.88h,AUC=98.656(ng/ml)*h. It was shown in pictures of SPECT, if NGF was packed with liposome there was NGF distributing in brain. Analysis of urine and dejecta indicated that main excretion route of unpacked NGF was urine and that of NGF packed with liposome was urine as well bile. Detection of NGF in CSF pointed that if NGF was packed peak of concentration in CSF was at 2hours whereas if NGF was unpacked NGF was undetectable. From SPECT, it was shown that there was radiological concentration in brain if 99mTc-NGF was packed with liposome. Radioactivity of urine deposit was u
ndetectable in TCA method.
Conclusion
When NGF is packed with liposome, absorption, distribution, metabolism and excretion of NGF will change. T1/2 and T1/2p are prolonged and NGF can pass BBB.
引文
1. Weihong Pan, William A. Banks, Abba J. Kastin. Permeability of the blood- brain barrier to neurotrophins. Brain Research 788 1998: 87～94
2. Thoenen H, Barde Y-A, Physiology of growth factor. Physiological Rev, 1980,60(4):1284-1335
3. Bradshaw RA. Nerve growth factor. Ann Rev Bilchem, 1978, 47:191-216
4. Marius C. Hoener, Eleanore Hewitt, James M. Conner, James W. Costello, Silvio Varon. Nerve growth factor (NGF) content in adult rat brain tissues is several-fold higher than generally reported and is largely associated with sedimentable fractions. Brain Research 728 1996:47-56
5. G. Vantini, S. D. Skaper. Liposome targeting to mouse brain: mannose as a recognition marker.Biochemical and biophysical Res Communication, 1998,153(3): 1038-1044
7. Ruben J. Boado, Haruhisa Tsukamoto, William. Paroride Drugs Delivery of Antisense Molecules to the brain for treatment of Alzheimer's disease and cerebral Aids .Journal of Pharmaceutical Sciences 1998, 87(11): 1308-1315
8. Ikumi Tanai, Akira Tsuji. Transpoter-Mediated permeation of Drugs across the BBB. J. Pharmaceutical Sciences, 2000, 89 (11): 1371-1888
9. M. Catherine Gustilo, Alicja L. Markowska. Evidence That Nerve Growth Factor Influences Recent Memory Through Structural Changes in Septohippocampal Cholinergic Neurons. The Journal Of Comparative Neurology 1999,405:491～507
10. Joseph F.Poduslo, Geoffrey L.Curran. Permeability at the blood-brain and blood-nerve barriers or the nertrophic factors: NGF,
CNTF, NF3, BDNF. Molecular Brain Res, 1999, (36): 280-286
11. Horikawa-I, Miwa-T TrkA expression in olfactory epithelium and bulb during development, Neuroport. 1999 Jul 13:10(10):2205-8.
12. Miwa-T, Retrograde transport of NGF from olfactory bulb to olfactory epithelium, Neuroreport, 1998,9(1):153-5
13. G.Vantini, S.D.Skaper. Innovations in drug delivery to the center nerve system. Drug of Today, 1999,35(6) :435-448
14. Maria Antonetta Tria, Maria Fusco, Guido Vantini etc. Controlled release of NGF from a polymeric impact. Brain Res, 1990, (515): 309-311
15. Tony L. Yaksh, Jongdae Jang, Yukiko Nishiuchi. The utility of 2-hydroxypropyl-β-cyclodextrin as a vehicle for the intracerebral and intratheral administration of drugs. Life Sciences, 1999,48
16. Suider W D. Functions of the neurotrophins during nerucells system development: what the knockouts are teaching US. Cell, 1994; 77:627
17. Levi-Montalicini R. The nerve growth factor thirty-five years later. Science, 1987; 237(4819):1154～1162
18. Ullish A Gray a. et al.Human beta NGF sequence highly homologous to that of mouse. Nature, 1983,303(3):821～825
19. Levi-Montalicini R, Meyer H, Hamburger V. In vitro experiments on the effects of mouse Sarcomas 180 and 37 on the spinal and sympathetic ganglia of the chick embryo. Cancer Res., 1954, 14(1):49
20. Tiscbler AS, Green L A. Nerve growth factor- induced process formation by cultured rat pheochromocytolna cells. Nature, 1975, 258(5333):341
21. Waterfield MD. Growth factors-Introduction. Brit Med Bull, 1989;45(2): 317
22. Bradford M M. A rapid and sensitive method for the quantitation of microgram of protein utilizing the principle of protein dye binding [J]. Anal BIOCHEM,1976,72:248-254.
23. Dryer R L, Lata G F. Experimental biochemistry [M]. New York: Oxford Yniversity Press, 1989:346-347.
24. Lowey O H, Rosebrough N J, Farr A L, et al. Protein measurement with the Folin phenol reagent. J Biol Chem,1951,193:265-269
25. Laemmli UK, Beguin F, Gujer- Kellenberger G. Cleavage of Structual proteins during the assembly of the head of bacteriophage T4. Nature, 1970, 227(5259): 680～685
26. Bangham AD, Standish M M, Watkins J C. Diffusion of univalent inos across the lamella of swollen phospholipids. J. Mol. Biol. 1965, 13:238～252.
27. Ng KY, Zhao L, Liu Y, et al. The effects ofpolyethyleneeglycol (PEG)-derived lipid on the activity of target-sensitive immunoliposome. Int J Pharm, 2000, 193(2):157～166
28. Xiaoyun Xua, Hanry Yua, Shujun Gaoa, Hai-Quan Maob, Kam W. Leong,Shu Wanga. Polyphosphoester microspheres for sustained release of biologically active nerve growth factor. Biomaterials 23 (2002)
29. Szoka F J, Papahadjopoulos D. Proc. Natl. Acad. Sci. USA, 1978, 75:4194～4198
30. Tria M A,Fusoo M. Pharmacokinetics of nerve growth factor (NGF) following different routes of administration to adult rats. Experimental Neurology, 1994;127:178～83
31. TANG Gang-hua, WANG Mingfang, TANG Xiao-Lan,et al Pharrnacokinetics and Radiation Dosimetry of O"(2-[18 F] fluoroethyl-L- Tyrosine as Oncologic PET Tracer. Int J Appl Radiat Isot,2003,
58(2):219-225.
32. Gupta PK. Quantitive evaluation of targeted drug delivery system. Int J Pharm, 1989, 56:27-34
33. Angeletti R H, Angeletti P U, Levi-Montalcini R. Brain Res, 1972,46:421-425
34. Tria M A, Fusco M, Vantini G, et al. Exp Neurol, 1994,127:178-183
35. Lawrince F, et al.Science, 1986, 235:214
36．王国华 陈南春 惠宏襄．小鼠颌下腺神经生长因子的制备和活性鉴定．第四军医大学学报，1998，19(6)：646～648
37．黄秉仁，蔡良琬．NGF家族及其受体研究进展．生理科学进展，1995，26(2)：115-120
38．丁玄宙．1986年诺理尔生理学或医学奖：神经生长因子和表皮生长因子的发现．生理科学进展，1987，18(2)：191
39．蔡文琴，李海标．发育神经生物学．北京：科学出版社，2001．76-97．142-174．
40．王占友，石玉秀．神经生长因子与运动神经再生[J]．解剖科学进展，1997,3(2)：126—129．
41．柳川．糖尿病周围神经病与 NGF．Foreign Medical Sciences section on pharmacy,1998．(788)：87-94
42．姜国辉，唐刚华，张云．神经生长因子的药代动力学研究．中国药学杂志，2000，35(3)：181～184
43．赵志刚．血脑屏障及药物通过BBB方法研究进展．中国药学杂志，2000，35(4)：227-230
44．王凤山、张天民、史群等神经生长因子及其研究概况 氨基酸杂志 1990(3)：36
45．张灵芝、凌世长、赵晶等．脂质体导向给药治疗心脑血管疾病的实验研究．北京医科大学学报，1994,vol26,：增刊：144～152
46．邵景范，罗永湘．神经生长因子的载体及其控释系统．中华显微外科杂志．1995，18(4)：303-305
47．平其能．现代药剂学．中国医药科技出版社，北京．1998年第1版：594～604
48．苏振刚，曹晓建，陈骐．国外医学脑血管疾病分册．2003，11(4)：290～293
49．柳川，李晋萍，华仲慰等．小鼠颌下腺2．5S神经生长因子的纯化和活性鉴定．军事医学科学院院刊，1 997,(11)2：28．
50．李素芹，李法卿，谭维国．人神经生长因子的规模纯化．中国生化药物杂志．1999，20(2)：123～125
51．董纯定．生化药物分析．中国药科大学出版社，南京．1995：142-150．
52．邹一愚，顾学裘，崛越勇。肝动脉注射阿霉素温度敏感脂质体的制剂研究[J]．药学学报，1991，26(8)：622～626．
53．张青，朱家壁．脂质体药物传输系统的研究进展．药学进展，2004，28(2)：68～71
54．谢溱，沈心亮，辛芳．ELISA法对神经生长因子在大鼠体内药代动力学的研究．中国药理学通报，1999，15(3)：268～70
55．唐刚华．正电子发射断层显像与新药研究[J]．药学学报，2001，36(6)：470—474
56．王世真，林俊．核医学与核技术(基础和应用)．科学出版社，北京．1990，第一版：366～372
57．李永健，孙其薰．放射性药物与标记化合物．科学出版社，北京．1996，第一版：40～45
58．朱桐，叶德华．放射性锝标记药物．见：马寄晓，刘秀杰．实用临床核医学．第一版．北京：原子能出版社，1990．54-55
59．常桂民、段芳龄、盛剑秋．半乳糖脂修饰的脂质体靶肝特性实验研究．中华核医学杂志．1998，18(4)：256～258
60．姜国辉，唐刚华，陈力真．神经生长因子在小鼠体内的分布与排泄实验研究．核技术．1998，21(11)：649～652
61．汤仲明，刘秀文，柴彪新．蛋白质多肽类药物药代动力学研究的方法学和实验设计．中国药理学与毒理学杂志 1996，10(3)：161～168
2. Thoenen H, Barde Y-A, Physiology of growth factor. Physiological Rev, 1980,60(4):1284-1335
3. Bradshaw RA. Nerve growth factor. Ann Rev Bilchem, 1978, 47:191-216
4. Marius C. Hoener, Eleanore Hewitt, James M. Conner, James W. Costello, Silvio Varon. Nerve growth factor (NGF) content in adult rat brain tissues is several-fold higher than generally reported and is largely associated with sedimentable fractions. Brain Research 728 1996:47-56
5. G. Vantini, S. D. Skaper. Liposome targeting to mouse brain: mannose as a recognition marker.Biochemical and biophysical Res Communication, 1998,153(3): 1038-1044
7. Ruben J. Boado, Haruhisa Tsukamoto, William. Paroride Drugs Delivery of Antisense Molecules to the brain for treatment of Alzheimer's disease and cerebral Aids .Journal of Pharmaceutical Sciences 1998, 87(11): 1308-1315
8. Ikumi Tanai, Akira Tsuji. Transpoter-Mediated permeation of Drugs across the BBB. J. Pharmaceutical Sciences, 2000, 89 (11): 1371-1888
9. M. Catherine Gustilo, Alicja L. Markowska. Evidence That Nerve Growth Factor Influences Recent Memory Through Structural Changes in Septohippocampal Cholinergic Neurons. The Journal Of Comparative Neurology 1999,405:491～507
10. Joseph F.Poduslo, Geoffrey L.Curran. Permeability at the blood-brain and blood-nerve barriers or the nertrophic factors: NGF,
CNTF, NF3, BDNF. Molecular Brain Res, 1999, (36): 280-286
11. Horikawa-I, Miwa-T TrkA expression in olfactory epithelium and bulb during development, Neuroport. 1999 Jul 13:10(10):2205-8.
12. Miwa-T, Retrograde transport of NGF from olfactory bulb to olfactory epithelium, Neuroreport, 1998,9(1):153-5
13. G.Vantini, S.D.Skaper. Innovations in drug delivery to the center nerve system. Drug of Today, 1999,35(6) :435-448
14. Maria Antonetta Tria, Maria Fusco, Guido Vantini etc. Controlled release of NGF from a polymeric impact. Brain Res, 1990, (515): 309-311
15. Tony L. Yaksh, Jongdae Jang, Yukiko Nishiuchi. The utility of 2-hydroxypropyl-β-cyclodextrin as a vehicle for the intracerebral and intratheral administration of drugs. Life Sciences, 1999,48
16. Suider W D. Functions of the neurotrophins during nerucells system development: what the knockouts are teaching US. Cell, 1994; 77:627
17. Levi-Montalicini R. The nerve growth factor thirty-five years later. Science, 1987; 237(4819):1154～1162
18. Ullish A Gray a. et al.Human beta NGF sequence highly homologous to that of mouse. Nature, 1983,303(3):821～825
19. Levi-Montalicini R, Meyer H, Hamburger V. In vitro experiments on the effects of mouse Sarcomas 180 and 37 on the spinal and sympathetic ganglia of the chick embryo. Cancer Res., 1954, 14(1):49
20. Tiscbler AS, Green L A. Nerve growth factor- induced process formation by cultured rat pheochromocytolna cells. Nature, 1975, 258(5333):341
21. Waterfield MD. Growth factors-Introduction. Brit Med Bull, 1989;45(2): 317
22. Bradford M M. A rapid and sensitive method for the quantitation of microgram of protein utilizing the principle of protein dye binding [J]. Anal BIOCHEM,1976,72:248-254.
23. Dryer R L, Lata G F. Experimental biochemistry [M]. New York: Oxford Yniversity Press, 1989:346-347.
24. Lowey O H, Rosebrough N J, Farr A L, et al. Protein measurement with the Folin phenol reagent. J Biol Chem,1951,193:265-269
25. Laemmli UK, Beguin F, Gujer- Kellenberger G. Cleavage of Structual proteins during the assembly of the head of bacteriophage T4. Nature, 1970, 227(5259): 680～685
26. Bangham AD, Standish M M, Watkins J C. Diffusion of univalent inos across the lamella of swollen phospholipids. J. Mol. Biol. 1965, 13:238～252.
27. Ng KY, Zhao L, Liu Y, et al. The effects ofpolyethyleneeglycol (PEG)-derived lipid on the activity of target-sensitive immunoliposome. Int J Pharm, 2000, 193(2):157～166
28. Xiaoyun Xua, Hanry Yua, Shujun Gaoa, Hai-Quan Maob, Kam W. Leong,Shu Wanga. Polyphosphoester microspheres for sustained release of biologically active nerve growth factor. Biomaterials 23 (2002)
29. Szoka F J, Papahadjopoulos D. Proc. Natl. Acad. Sci. USA, 1978, 75:4194～4198
30. Tria M A,Fusoo M. Pharmacokinetics of nerve growth factor (NGF) following different routes of administration to adult rats. Experimental Neurology, 1994;127:178～83
31. TANG Gang-hua, WANG Mingfang, TANG Xiao-Lan,et al Pharrnacokinetics and Radiation Dosimetry of O"(2-[18 F] fluoroethyl-L- Tyrosine as Oncologic PET Tracer. Int J Appl Radiat Isot,2003,
58(2):219-225.
32. Gupta PK. Quantitive evaluation of targeted drug delivery system. Int J Pharm, 1989, 56:27-34
33. Angeletti R H, Angeletti P U, Levi-Montalcini R. Brain Res, 1972,46:421-425
34. Tria M A, Fusco M, Vantini G, et al. Exp Neurol, 1994,127:178-183
35. Lawrince F, et al.Science, 1986, 235:214
36．王国华 陈南春 惠宏襄．小鼠颌下腺神经生长因子的制备和活性鉴定．第四军医大学学报，1998，19(6)：646～648
37．黄秉仁，蔡良琬．NGF家族及其受体研究进展．生理科学进展，1995，26(2)：115-120
38．丁玄宙．1986年诺理尔生理学或医学奖：神经生长因子和表皮生长因子的发现．生理科学进展，1987，18(2)：191
39．蔡文琴，李海标．发育神经生物学．北京：科学出版社，2001．76-97．142-174．
40．王占友，石玉秀．神经生长因子与运动神经再生[J]．解剖科学进展，1997,3(2)：126—129．
41．柳川．糖尿病周围神经病与 NGF．Foreign Medical Sciences section on pharmacy,1998．(788)：87-94
42．姜国辉，唐刚华，张云．神经生长因子的药代动力学研究．中国药学杂志，2000，35(3)：181～184
43．赵志刚．血脑屏障及药物通过BBB方法研究进展．中国药学杂志，2000，35(4)：227-230
44．王凤山、张天民、史群等神经生长因子及其研究概况 氨基酸杂志 1990(3)：36
45．张灵芝、凌世长、赵晶等．脂质体导向给药治疗心脑血管疾病的实验研究．北京医科大学学报，1994,vol26,：增刊：144～152
46．邵景范，罗永湘．神经生长因子的载体及其控释系统．中华显微外科杂志．1995，18(4)：303-305
47．平其能．现代药剂学．中国医药科技出版社，北京．1998年第1版：594～604
48．苏振刚，曹晓建，陈骐．国外医学脑血管疾病分册．2003，11(4)：290～293
49．柳川，李晋萍，华仲慰等．小鼠颌下腺2．5S神经生长因子的纯化和活性鉴定．军事医学科学院院刊，1 997,(11)2：28．
50．李素芹，李法卿，谭维国．人神经生长因子的规模纯化．中国生化药物杂志．1999，20(2)：123～125
51．董纯定．生化药物分析．中国药科大学出版社，南京．1995：142-150．
52．邹一愚，顾学裘，崛越勇。肝动脉注射阿霉素温度敏感脂质体的制剂研究[J]．药学学报，1991，26(8)：622～626．
53．张青，朱家壁．脂质体药物传输系统的研究进展．药学进展，2004，28(2)：68～71
54．谢溱，沈心亮，辛芳．ELISA法对神经生长因子在大鼠体内药代动力学的研究．中国药理学通报，1999，15(3)：268～70
55．唐刚华．正电子发射断层显像与新药研究[J]．药学学报，2001，36(6)：470—474
56．王世真，林俊．核医学与核技术(基础和应用)．科学出版社，北京．1990，第一版：366～372
57．李永健，孙其薰．放射性药物与标记化合物．科学出版社，北京．1996，第一版：40～45
58．朱桐，叶德华．放射性锝标记药物．见：马寄晓，刘秀杰．实用临床核医学．第一版．北京：原子能出版社，1990．54-55
59．常桂民、段芳龄、盛剑秋．半乳糖脂修饰的脂质体靶肝特性实验研究．中华核医学杂志．1998，18(4)：256～258
60．姜国辉，唐刚华，陈力真．神经生长因子在小鼠体内的分布与排泄实验研究．核技术．1998，21(11)：649～652
61．汤仲明，刘秀文，柴彪新．蛋白质多肽类药物药代动力学研究的方法学和实验设计．中国药理学与毒理学杂志 1996，10(3)：161～168
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |