3株Lactobacillus rhamnosus的分子鉴定及其对Wistar大鼠血清胆固醇含量的影响
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摘要
为在分子水平上鉴定3株乳酸菌,探究其对Wistar大鼠血清胆固醇含量的影响。试验对菌株D-3、D-12和A-5进行16 S rDNA鉴定,64只大鼠随机分空白对照组、高能对照组、普通试验组(普通1组、普通2组、普通3组)和高能试验组(高能1组、高能2组、高能3组);空白对照组与普通试验组饲喂普通饲料、高能对照组与高能试验组饲喂高能饲料,高脂模型建立成功后,空白对照组、高能对照组灌胃生理盐水,普通试验组与高能试验组分别依次灌胃D-3、D-12和A-5菌悬液,于第7天、第21天采血,用单试剂法测定其血清中总胆固醇(Total Cholesterol,TC)、甘油三酯(Triglyceride,TG)含量;21 d后摘取肝脏称重,测算其指数。结果:3株菌均鉴定为鼠李糖乳杆菌(Lactobacillus rhamnosus);普通试验组与空白对照组大鼠血清中TC、TG含量均无显著差异(P>0.05);高能试验组TC、TG含量显著低于高能对照组(P<0.05)高于空白对照组(P<0.05);第21天时,高能2组TG含量均显著低于高能1组与3组(P<0.05),TC含量显著低于高能1组(P<0.05)略低于高能3组(P>0.05);普通试验组肝脏指数略低于空白对照组(P>0.05),高能试验组肝脏指数均显著低于高能对照组高于空白对照组(P<0.05),高能2组肝脏指数均显著低于高能1组与3组(P<0.05)。结论:乳酸菌D-3、D-12和A-5都属鼠李糖乳杆菌,均对高脂Wistar大鼠血清TC、TG具有良好降低作用,且D-12降低效果最佳。
The study was aimed to identify three strains of lactic acid bacteria, at the molecular level, and research the effect of its influence on blood cholesterol, in Wistar rats. The strains D-3, D-12, and A-5 were identified by the 16 S rDNA molecularly method, 64 Wistar rats were divided into blank control group, high energy control group, normal test group(group 1, group 2, group 3) and high energy test group(group 1, group 2, group 3), 8 per group, randomly. The control and normal group were fed with normal feeds; high energy control and high-energy group test were feel high-energy feed to establish high fat mold. Let it continue to eat freely after model succeeds. The blank control and high energy control were given normal saline, and the normal and high energy test group were given with D-3, D-12, and A-5. The blood was collected, on the 7 th day and the 21 st day. The serum total cholestero(TC) and triglyceride(TG) levels were measured by the single-agent method; The rat's complete liver were removed and weight after the 21 st day, calculating the index. The results showed that 3 strains was Lactobacillus rhamnosus; The normal test and blank control group serum TC and TG content was not significantly different(P>0.05); High test group serum TC, TG were significantly lower than the high control group than the normal control group(P<0.05); High energy group 2 serum TG contents was significantly lower than high energy group 1 and group 3(p<0.05); High energy group 2 serum TC was significantly lower than the high-energy group 1(P<0.05), but slightly lower than the high energy groups 3(P>0.05), on the the 21 st day; Normal test group liver index was lower than blank control group, slightly(P>0.05); High energy test group liver index were significantly lower than high energy control group, and significantly than the control group(P<0.05); The high energy group 2 liver index were significantly lower than high energy group 1 and group 3(P<0.05), in high energy group. Based on the above, Lactobacillus D-3, D-12 and A-5 have a good effect on the blood cholesterol of high-fat Wistar rats, which belong to Lactobacillus rhamnosus, and Lactobacillus rhamnosus D-12 has the best effect on blood cholesterol reduction.
The study was aimed to identify three strains of lactic acid bacteria, at the molecular level, and research the effect of its influence on blood cholesterol, in Wistar rats. The strains D-3, D-12, and A-5 were identified by the 16 S rDNA molecularly method, 64 Wistar rats were divided into blank control group, high energy control group, normal test group(group 1, group 2, group 3) and high energy test group(group 1, group 2, group 3), 8 per group, randomly. The control and normal group were fed with normal feeds; high energy control and high-energy group test were feel high-energy feed to establish high fat mold. Let it continue to eat freely after model succeeds. The blank control and high energy control were given normal saline, and the normal and high energy test group were given with D-3, D-12, and A-5. The blood was collected, on the 7 th day and the 21 st day. The serum total cholestero(TC) and triglyceride(TG) levels were measured by the single-agent method; The rat's complete liver were removed and weight after the 21 st day, calculating the index. The results showed that 3 strains was Lactobacillus rhamnosus; The normal test and blank control group serum TC and TG content was not significantly different(P>0.05); High test group serum TC, TG were significantly lower than the high control group than the normal control group(P<0.05); High energy group 2 serum TG contents was significantly lower than high energy group 1 and group 3(p<0.05); High energy group 2 serum TC was significantly lower than the high-energy group 1(P<0.05), but slightly lower than the high energy groups 3(P>0.05), on the the 21 st day; Normal test group liver index was lower than blank control group, slightly(P>0.05); High energy test group liver index were significantly lower than high energy control group, and significantly than the control group(P<0.05); The high energy group 2 liver index were significantly lower than high energy group 1 and group 3(P<0.05), in high energy group. Based on the above, Lactobacillus D-3, D-12 and A-5 have a good effect on the blood cholesterol of high-fat Wistar rats, which belong to Lactobacillus rhamnosus, and Lactobacillus rhamnosus D-12 has the best effect on blood cholesterol reduction.
引文
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[12]Guo Chunfeng,Zhang Shuang,Yuan Yahong,et al.Comparison of lactobacilli isolated from Chinese suantsai and koumiss for their probiotic and functional properties[J].Journal of Functional Foods,2015,12(2):294-302
[13]卢海鹏,赵俊良,刘景波,等.长期喂服酸马奶对Wistar大鼠外周血淋巴细胞数量和百分比的影响[J].细胞与分子免疫学杂志,2017,33(8):1087-1090
[14]黄德强.大鼠高脂模型的建立[J].中国畜牧业,2016,(11):58-59
[15]EI-GAWAD I A A,EI-SAYED E M,HAFEZ S A,et al.The Hypocholesterolaemic Effect of Milk Yoghurt and Soy-yoghurt Containing Bifidobacteria in Rats Fed on a Cholesterol-enriched Diet[J].International Dairy Journal,2005,15(1):37-44
[16]KUMAR R,GROVER S,BATISH V.Bile salt hydrolase activity screening of lactobacilli:in vitro selection of indigenous Lactobacillus strains with potential bile salt hydrolysing and cholesterol-lowering ability[J].Probiotics&Antimicrobial Proteins,2012,4(3):162-172
[17]WRIGHT R S,ANDERSON J W,BRIDGES S R.Propionate inhibits hepatocyte lipid synthesis[J].Experimental Biology and Medicine,1990,195(1):26-29
[18]Al-Sheraji,Sadeq Hasan,et al.Hypocholesterolaemic e f f e c t o f y o g h u r t c o n t a i n i n g B i f i d o b a c t e r i u m Pseudocatenulatum G4 or Bifidobacterium longum BB536[J].Food Chemistry,2012,135(2):356-361
[19]Chiu H C,Tzu Y L,Yun YT.The effects of Lactobacillusfermented milk on lipid metabolism in hamsters fed on high-cholesterol diet[J].Applied Microbiology and Biotechnology,2006,71(2):238-245
[20]Fazeli H,Moshtaghian J,Mir-lohii M,et al.Reduction in Serum Lipid Parameters by Incorporation of a Native Strain of Lactobacillus Plantarum A7 in Mice[J].Iranian Journal of Diabetes and Lipid Disorders,2010,9(9):1-7
[2]Kapourchali F R,Surendiran G,Goulet A,et al.The role of dietary cholesterol in lipoprotein metabolism and related metabolic abnormalities:A Mini-review[J].Crit Rev Food Sci Nutr,2016,56(14):2408-2415
[3]Borzyszkowska J,Stanislawskasachadyn A,Wirtwein M,et al.Angiotensin converting enzyme gene polymorphism is associated with severity of coronary artery disease in men with high total cholesterol levels[J].Journal of applied genetics,2012,53(2):175-182
[4]Yung Liang C,Liang Yi W,Tsung Cheng L,et al.Cholesterol-lowering effect of phytosterol-containing lactic-fermented milk powder in hamsters[J].Food Chemistry,2010,119(3):1121-1126
[5]MANN G V,SPOERR Y A.Study of surfactant and cholsteremia in the Massi[J].American Journal of Clinical Nutrition,1974,27:464-469
[6]Zhang Jiachao,Wang Lifeng,Zhuang Guo,et al.454pyrosequencing reveals changes in the faecal microbiota of adults consuming Lactobacillus casei Zhang[J].Fems Microbiology Ecology,2014,88(3):6-12
[7]王秀娟,张小磊,商雪娇,等.2种乳酸菌降胆固醇能力的比较研究[J].食品科技,2016,41(05):12-15
[8]于志会,张雪,李盛钰,等.植物乳杆菌C88发酵乳对高脂血症小鼠降胆固醇的作用[J].中国乳品工业,2013,41(8):17-19
[9]韦云路,刘义,王瑶,等.3株益生菌体外降胆固醇能力及体内降血脂效果评价[J].食品科学,2017,38(23):129-134
[10]毕斯哈拉图.蒙药当玛-5酸马奶咀嚼片用于功能性消化不良大鼠胃排空中的效果[J].中国民族医药杂志,2017,23(4):49-50
[11]赵俊良,卢海鹏,金山,等.乳酸菌盐溶性蛋白培养物中血管紧张素转化酶抑制活性肽的测定及分离[J].食品科学,2016,37(9):170-174
[12]Guo Chunfeng,Zhang Shuang,Yuan Yahong,et al.Comparison of lactobacilli isolated from Chinese suantsai and koumiss for their probiotic and functional properties[J].Journal of Functional Foods,2015,12(2):294-302
[13]卢海鹏,赵俊良,刘景波,等.长期喂服酸马奶对Wistar大鼠外周血淋巴细胞数量和百分比的影响[J].细胞与分子免疫学杂志,2017,33(8):1087-1090
[14]黄德强.大鼠高脂模型的建立[J].中国畜牧业,2016,(11):58-59
[15]EI-GAWAD I A A,EI-SAYED E M,HAFEZ S A,et al.The Hypocholesterolaemic Effect of Milk Yoghurt and Soy-yoghurt Containing Bifidobacteria in Rats Fed on a Cholesterol-enriched Diet[J].International Dairy Journal,2005,15(1):37-44
[16]KUMAR R,GROVER S,BATISH V.Bile salt hydrolase activity screening of lactobacilli:in vitro selection of indigenous Lactobacillus strains with potential bile salt hydrolysing and cholesterol-lowering ability[J].Probiotics&Antimicrobial Proteins,2012,4(3):162-172
[17]WRIGHT R S,ANDERSON J W,BRIDGES S R.Propionate inhibits hepatocyte lipid synthesis[J].Experimental Biology and Medicine,1990,195(1):26-29
[18]Al-Sheraji,Sadeq Hasan,et al.Hypocholesterolaemic e f f e c t o f y o g h u r t c o n t a i n i n g B i f i d o b a c t e r i u m Pseudocatenulatum G4 or Bifidobacterium longum BB536[J].Food Chemistry,2012,135(2):356-361
[19]Chiu H C,Tzu Y L,Yun YT.The effects of Lactobacillusfermented milk on lipid metabolism in hamsters fed on high-cholesterol diet[J].Applied Microbiology and Biotechnology,2006,71(2):238-245
[20]Fazeli H,Moshtaghian J,Mir-lohii M,et al.Reduction in Serum Lipid Parameters by Incorporation of a Native Strain of Lactobacillus Plantarum A7 in Mice[J].Iranian Journal of Diabetes and Lipid Disorders,2010,9(9):1-7
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