乳酸菌对高糖高脂2型糖尿病小鼠血脂的影响
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摘要
目的:研究两株乳酸菌对糖尿病小鼠血糖、血脂、血清炎症因子、肝脏抗氧化水平、胰腺病理变化和附睾脂肪组织白细胞介素-10(interleukin-10,IL-10)及脂联素(adiponectin,Adipoq)基因表达的影响。方法:通过降胆固醇、耐酸及耐胆盐等体外实验从6株乳酸菌中筛选降脂较好的2株乳酸菌。利用链脲佐菌素(streptozotocin,STZ)结合高糖高脂饲料的方法建立小鼠糖尿病模型。将100只雄性BALB/c小鼠随机分为4组,每组25只,对照组(N)(正常饮食)、模型组(M)(高糖高脂饮食)、SY13菌组(SY13)(高糖高脂饮食外加SY13菌液灌胃)、36号菌组(D36)(高糖高脂饮食外加36号菌液灌胃),分别在灌胃6、10周和14周时处死小鼠,动态检测小鼠血糖、血脂等相关生化指标水平,利用苏木精-伊红染色观察胰腺组织病理变化,通过逆转录实时荧光定量聚合酶链式反应方法检测附睾脂肪组织中IL-10和Adipoq基因的m RNA表达情况。结果:筛选出干酪乳酸菌SY13和D36有较强的降胆固醇、耐酸和耐胆盐能力。与M组相比,通过一段时间菌液的灌胃,显著抑制了小鼠血糖水平和血清中总胆固醇、甘油三酯、低密度脂蛋白胆固醇水平的上升(P<0.05),提高了高密度脂蛋白胆固醇水平和抗炎因子IL-10水平(P<0.05),不同程度提高了肝脏谷胱甘肽过氧化物酶、超氧化物歧化酶和过氧化氢酶活力,显著降低了丙二醛含量(P<0.05),改善了小鼠糖代谢紊乱状况,明显减轻了胰岛和腺泡细胞病变情况,脂肪组织中IL-10和Adipoq基因表达水平显著提高。结论:乳酸菌SY13和D36能够显著改善糖高脂饮食联合STZ诱导的小鼠糖脂代谢。
Objective: To study the effects of two Lactobacillus strains on blood glucose, blood lipids, serum inflammatory cytokines, liver antioxidant level, pancreatic pathology, and the expression of the genes encodig interleukin-10(IL-10)and adiponectin(adipoq) in epididymal adipose tissue in diabetic mice. Methods: Out of six Lactobacillus strains, two Lactobacillu casei strains(SY13 and D36) were selected for their potent lipid-lowering effect in terms of cholesterollowering ability, and acid and bile salt tolerance in vitro. The diabetic mouse model was established by feeding mice a highglucose and high-fat diet and injecting them with streptozotocin(STZ). Totally 100 male Balb/c mice were randomly divided into four groups of 25 mice per group: control, model, SY13 and D36 groups. The control group was fed a normal diet, while all the other groups were given a high-glucose and high-fat diet. The SY13 and D36 groups were additionally administered by gavage with Lactobacillu casei SY13 and D36, respectively. Some of the mice were scarified at the end of the sixth, tenth and fourth week of administration for the detection of blood glucose, blood lipids and other related biochemical indexes. HE staining was used to observe the pathological change of pancreatic tissues, and the mRNA expression of IL-10 and Adipoq genes in epididymal adipose tissues were detected by reverse transcription quantative real-time polymerase chain reaction.Results: L. casei SY13 and D36 had high cholesterol-lowering capacity, and strong acid and bile salt tolerance. Compared with the model group, both strains significantly repressed the increase in blood sugar and serum total cholesterol, triglyceride and low-density lipoprotein cholesterol levels, and increased the levels of serum high-density lipoprotein chrolestserol and anti-inflammatory factor interleukin-10(IL-10)(P < 0.05). In addition, they significantly increased the activity of glutatione peroxidase, superoxide dismutase and catalase in liver and decreased malondialdehyde(P < 0.05). The strains also improved glucose metabolism disorders, obviously alleviated the pathological changes of islet and adenocytic cells, and promoted the mRNA expression of IL-10 and Adipoq genes in adipose tissues. Conclusion: Lactobacillus casei SY13 and D36 can significantly improve glucose and lipid metabolism in mice with diabetes induced by a high-glucose and high-fat diet combined with STZ injection.
Objective: To study the effects of two Lactobacillus strains on blood glucose, blood lipids, serum inflammatory cytokines, liver antioxidant level, pancreatic pathology, and the expression of the genes encodig interleukin-10(IL-10)and adiponectin(adipoq) in epididymal adipose tissue in diabetic mice. Methods: Out of six Lactobacillus strains, two Lactobacillu casei strains(SY13 and D36) were selected for their potent lipid-lowering effect in terms of cholesterollowering ability, and acid and bile salt tolerance in vitro. The diabetic mouse model was established by feeding mice a highglucose and high-fat diet and injecting them with streptozotocin(STZ). Totally 100 male Balb/c mice were randomly divided into four groups of 25 mice per group: control, model, SY13 and D36 groups. The control group was fed a normal diet, while all the other groups were given a high-glucose and high-fat diet. The SY13 and D36 groups were additionally administered by gavage with Lactobacillu casei SY13 and D36, respectively. Some of the mice were scarified at the end of the sixth, tenth and fourth week of administration for the detection of blood glucose, blood lipids and other related biochemical indexes. HE staining was used to observe the pathological change of pancreatic tissues, and the mRNA expression of IL-10 and Adipoq genes in epididymal adipose tissues were detected by reverse transcription quantative real-time polymerase chain reaction.Results: L. casei SY13 and D36 had high cholesterol-lowering capacity, and strong acid and bile salt tolerance. Compared with the model group, both strains significantly repressed the increase in blood sugar and serum total cholesterol, triglyceride and low-density lipoprotein cholesterol levels, and increased the levels of serum high-density lipoprotein chrolestserol and anti-inflammatory factor interleukin-10(IL-10)(P < 0.05). In addition, they significantly increased the activity of glutatione peroxidase, superoxide dismutase and catalase in liver and decreased malondialdehyde(P < 0.05). The strains also improved glucose metabolism disorders, obviously alleviated the pathological changes of islet and adenocytic cells, and promoted the mRNA expression of IL-10 and Adipoq genes in adipose tissues. Conclusion: Lactobacillus casei SY13 and D36 can significantly improve glucose and lipid metabolism in mice with diabetes induced by a high-glucose and high-fat diet combined with STZ injection.
引文
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[15]李向菲.产胞外多糖乳酸菌对2型糖尿病的干预及其机制研究[D].无锡:江南大学,2016:1-7.
[16]LI X F,WANG N,YIN B X,et al.Lactobacillus plantarum X1withα-glucosidase inhibitory activity ameliorates type 2 diabetes in mice[J].RSC Advances,2016,6(68):63536-63547.DOI:10.1039/C6RA10858J.
[17]LIONG M T,SHAH N P.Acid and bile tolerance and cholesterol removal ability of lactobacilli strains[J].Journal of Dairy Science,2005,88(1):55-66.DOI:10.3168/S0022-0302(05)72662-X.
[18]GRILL J P,CAYUELA C,ANTOINE J M,et al.Effects of Lactobacillus amylovorus and Bifidobacterium breve on cholesterol[J].Letters in Applied Microbiology,2000,31(2):154-156.
[19]KUMAR R,GROVER S,BATISH V K.Hypocholesterolaemic effect of dietary inclusion of two putative probiotic bile salt hydrolaseproducing Lactobacillus plantarum strains in Sprague-Dawley rats[J].British Journal of Nutrition,2011,105(4):561-73.DOI:10.1017/S0007114510003740.
[20]FUKUSHIMA M,NAKANO M.Effects of a mixture of organisms,Lactobacillus acidophilus or Streptococcus faecalis on cholesterol metabolism in rats fed on a fat-and cholesterol-enriched diet[J].British Journal of Nutrition,2007,76(6):857-867.DOI:10.1079/bjn19960092.
[21]郭均.降胆固醇乳酸菌的筛选及其体内降胆固醇作用[D].广州:华南理工大学,2016:17-18.
[22]DUNNE C,OMAHONY L,MURPHY L,et al.In vitro selection criteria for probiotic bacteria of human origin:correlation with in vivo findings[J].American Journal of Clinical Nutrition,2001,73(Suppl 2):386-392.
[23]于金慧,王瑜,彭振英,等.裂壶藻油DHA对高脂饮食诱导肥胖小鼠的影响[J].微生物学通报,2017,44(11):2679-2688.DOI:10.13344/j.microbiol.china.170355.
[24]谷豪,夏毅伟,韦莉萍,等.桃胶多糖、抗性淀粉、低聚果糖对糖耐量受损大鼠血糖的影响比较研究[J].热带医学杂志,2013,13(3):288-290.
[25]廖素凤,刘江洪,杨志坚,等.茶树新品系CFT-1提取物的降脂作用及其机制研究[J].天然产物研究与开发,2017,29(11):1831-1840;1857.
[26]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J].Methods,2001,25(4):402-408.DOI:10.1006/meth.2001.1262.
[27]ALP G,ASLIM B.Relationship between the resistance to bile salts and low pH with exopolysaccharide(EPS)production of Bifidobacterium spp.isolated from infants feces and breast milk[J].Anaerobe,2010,16(2):101-105.DOI:10.1016/j.anaerobe.2009.06.006.
[28]温贺,肖凤艳,段翠翠,等.植物乳杆菌Sc52益生特性评价及其在降血糖产品中的应用[J].食品科学,2018,39(6):148-154.DOI:10.7506/spkx1002-6630-201806024.
[29]李培培,黄庆德,许继取,等.部分脱脂亚麻籽粉对高脂高糖饮食大鼠血糖的影响[J].食品科学,2018,39(21):183-188.DOI:10.7506/spkx1002-6630-201821028.
[30]CAO H.Adipocytokines in obesity and metabolic disease[J].Journal of Endocrinology,2014,220(2):47-59.DOI:10.1530/JOE-13-0339.
[31]周敏.实验性2型糖尿病动物模型研究及其进展[J].浙江中医学院学报,2001(5):79-81.
[32]BOLLINGE S S,WELTMAN N Y,GERDES A M,et al.T3supplementation affects ventilatory timing&glucose levels in type 2diabetes mellitus model[J].Respiratory Physiology&Neurobiology,2015,205:92-98.DOI:10.1016/j.resp.2014.10.020.
[33]DANDONA P,ALJADA A,CHAUDHURI A,et al.Metabolic syndrome:a comprehensive perspective based on interactions between obesity,diabetes,and inflammation[J].Circulation,2005,111(11):1448-1454.DOI:10.1161/01.CIR.0000158483.13093.9D.
[2]马晖,谢春光.参芪复方降血糖实验研究[J].四川省卫生管理干部学院学报,2006(2):87-89.
[3]王芬,何华亮,张红敏,等.参芪复方对GK大鼠脂代谢异常的实验研究[J].天津中医药,2007(6):507-508.
[4]JEMIL I,ABDELHEDI O,NASRI R,et al.Hypolipidemic,antiobesity and cardioprotective effects of sardinelle meat flour and its hydrolysates in high-fat and fructose diet fed Wistar rats[J].Life Science,2017,176:54-66.DOI:10.1016/j.lfs.2016.07.012.
[5]孙康悦,李颖,李悦琪,等.国内降血脂药物研究进展[J].西北药学杂志,2017,32(1):122-124.
[6]陈大卫.辅助降血脂益生乳酸菌的筛选及其对高血脂大鼠肠道菌群的影响[D].扬州:扬州大学,2015:9-11.DOI:10.7666/d.Y2908625.
[7]李幼筠.泡菜与乳酸菌[J].中国酿造,2001,20(4):7-9.
[8]罗冬英,尹传武.乳酸菌制剂对人体保健功效的机理探讨[J].鄂州大学学报,2002(4):53-54.
[9]SMET I D,BOEVER P D,VERSTRAETE W.Cholesterol lowering in pigs through enhanced bacterial bile salt hydrolase activity[J].British Journal of Nutrition,2007,79(2):185-194.DOI:10.1079/bjn19980030.
[10]HUANG Y,ZHENG Y C.The probiotic Lactobacillus acidophilus reduces cholesterol absorption through the down-regulation of Niemann-Pick C1-like 1 in Caco-2 cells[J].British Journal of Nutrition,2009,103(4):473-478.DOI:10.1017/s0007114509991991.
[11]ALSHERAJI S H,ISMAIL A,MANAP M Y,et al.Hypocholesterolaemic effect of yoghurt containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536[J].Food Chemistry,2012,135(2):356-361.DOI:10.1016/j.foodchem.2012.04.120.
[12]JONES M L,MARTONI C J,PARENT M,et al.Cholesterol-lowering efficacy of a microencapsulated bile salt hydrolase-active Lactobacillus reuteri NCIMB 30242 yoghurt formulation in hypercholesterolaemic adults[J].British Journal of Nutrition,2012,107(10):1505-1513.DOI:10.1017/S0007114511004703.
[13]俞筱琦,施志冲,郑勇英,等.微生态类制剂调节血脂和润肠通便作用[J].上海实验动物科学,2001(2):109-110;112.
[14]VILAHUR G,LóPEZ-BERNAL S,CAMINO S,et al.Lactobacillus plantarum CECT 7315/7316 intake modulates the acute and chronic innate inflammatory response[J].European Journal of Nutrition,2015,54(7):1161-1171.DOI:10.1007/s00394-014-0794-9.
[15]李向菲.产胞外多糖乳酸菌对2型糖尿病的干预及其机制研究[D].无锡:江南大学,2016:1-7.
[16]LI X F,WANG N,YIN B X,et al.Lactobacillus plantarum X1withα-glucosidase inhibitory activity ameliorates type 2 diabetes in mice[J].RSC Advances,2016,6(68):63536-63547.DOI:10.1039/C6RA10858J.
[17]LIONG M T,SHAH N P.Acid and bile tolerance and cholesterol removal ability of lactobacilli strains[J].Journal of Dairy Science,2005,88(1):55-66.DOI:10.3168/S0022-0302(05)72662-X.
[18]GRILL J P,CAYUELA C,ANTOINE J M,et al.Effects of Lactobacillus amylovorus and Bifidobacterium breve on cholesterol[J].Letters in Applied Microbiology,2000,31(2):154-156.
[19]KUMAR R,GROVER S,BATISH V K.Hypocholesterolaemic effect of dietary inclusion of two putative probiotic bile salt hydrolaseproducing Lactobacillus plantarum strains in Sprague-Dawley rats[J].British Journal of Nutrition,2011,105(4):561-73.DOI:10.1017/S0007114510003740.
[20]FUKUSHIMA M,NAKANO M.Effects of a mixture of organisms,Lactobacillus acidophilus or Streptococcus faecalis on cholesterol metabolism in rats fed on a fat-and cholesterol-enriched diet[J].British Journal of Nutrition,2007,76(6):857-867.DOI:10.1079/bjn19960092.
[21]郭均.降胆固醇乳酸菌的筛选及其体内降胆固醇作用[D].广州:华南理工大学,2016:17-18.
[22]DUNNE C,OMAHONY L,MURPHY L,et al.In vitro selection criteria for probiotic bacteria of human origin:correlation with in vivo findings[J].American Journal of Clinical Nutrition,2001,73(Suppl 2):386-392.
[23]于金慧,王瑜,彭振英,等.裂壶藻油DHA对高脂饮食诱导肥胖小鼠的影响[J].微生物学通报,2017,44(11):2679-2688.DOI:10.13344/j.microbiol.china.170355.
[24]谷豪,夏毅伟,韦莉萍,等.桃胶多糖、抗性淀粉、低聚果糖对糖耐量受损大鼠血糖的影响比较研究[J].热带医学杂志,2013,13(3):288-290.
[25]廖素凤,刘江洪,杨志坚,等.茶树新品系CFT-1提取物的降脂作用及其机制研究[J].天然产物研究与开发,2017,29(11):1831-1840;1857.
[26]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method[J].Methods,2001,25(4):402-408.DOI:10.1006/meth.2001.1262.
[27]ALP G,ASLIM B.Relationship between the resistance to bile salts and low pH with exopolysaccharide(EPS)production of Bifidobacterium spp.isolated from infants feces and breast milk[J].Anaerobe,2010,16(2):101-105.DOI:10.1016/j.anaerobe.2009.06.006.
[28]温贺,肖凤艳,段翠翠,等.植物乳杆菌Sc52益生特性评价及其在降血糖产品中的应用[J].食品科学,2018,39(6):148-154.DOI:10.7506/spkx1002-6630-201806024.
[29]李培培,黄庆德,许继取,等.部分脱脂亚麻籽粉对高脂高糖饮食大鼠血糖的影响[J].食品科学,2018,39(21):183-188.DOI:10.7506/spkx1002-6630-201821028.
[30]CAO H.Adipocytokines in obesity and metabolic disease[J].Journal of Endocrinology,2014,220(2):47-59.DOI:10.1530/JOE-13-0339.
[31]周敏.实验性2型糖尿病动物模型研究及其进展[J].浙江中医学院学报,2001(5):79-81.
[32]BOLLINGE S S,WELTMAN N Y,GERDES A M,et al.T3supplementation affects ventilatory timing&glucose levels in type 2diabetes mellitus model[J].Respiratory Physiology&Neurobiology,2015,205:92-98.DOI:10.1016/j.resp.2014.10.020.
[33]DANDONA P,ALJADA A,CHAUDHURI A,et al.Metabolic syndrome:a comprehensive perspective based on interactions between obesity,diabetes,and inflammation[J].Circulation,2005,111(11):1448-1454.DOI:10.1161/01.CIR.0000158483.13093.9D.