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益生菌对慢加急性肝衰竭大鼠模型的保护作用及其机制
Protective effect of probiotics in rats with acute-on-chronic liver failure and related mechanism
文章发布日期:2019年06月14日  来源:  作者:张永超,毕研贞,方萧,等  点击次数:212次  下载次数:48次

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【摘要】:目的探究益生菌干预对慢加急性肝衰竭(ACLF)大鼠的作用及其机制。方法采用随机数字表法将44只雄性SD大鼠随机分为6组,对照组1(C1组,n=6,不做任何干预)、模型组1(M1组,n=8,40% CCl4油溶液腹腔注射10周,从第7周开始每天灌胃PBS溶液,10周末给予D-GalN急性攻击)、益生菌干预组1(Y1组,n=8,造模同M1组,灌胃剂为益生菌溶液)、对照组2(C2组,n=6,不做任何干预)、模型组2(M2组,n=8,腹腔注射40% CCl4 油溶液,10周后给予D-GalN急性攻击,48 h后每天灌胃PBS溶液,持续到第12周末处死)、益生菌干预组2(Y2组,n=8,造模同M2组,灌胃剂为益生菌溶液)。观察干预前后大鼠体质量、肝功能、肝组织病理学,采用ELISA法测血浆内毒素、肠道分泌型免疫球蛋白A(sIgA),Western Blot法测定肠道紧密连接蛋白occludin的水平,RT-PCR法检测肠道紧密连接蛋白occludin和ZO-1的mRNA水平,通过选择性培养基测定肠道菌群等指标的变化。计量资料多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果C1、M1、Y1 3组间与C2、M2、Y2 3组间体质量、肝指数、ALT、AST、TBil、血浆内毒素、sIgA、occludin mRNA、ZO-1mRNA水平比较差异均有统计学意义(F1值分别为2765、896、61.37、18.27、21.00、87.01、67.10、101.50、105.40,P值均<0.05;F2值分别为14.04、12.85、14.02、11.39、35.80、1914、1537、25.02、126.00,P值均<0.05),C1、M1、Y1 3组间occludin 蛋白水平比较差异有统计学意义(F=16.40,P<0.05)。C1、M1、Y1 3组间与C2、M2、Y2 3组间乳酸杆菌、双歧杆菌、肠球菌、肠杆菌含量比较差异均有统计学意义(F1分别为77.95、66.61、25.63、33.29,P值均<0.05;F2分别为21.50、22.62、6.71、17.74,P值均<0.05) 结论ACLF大鼠出现肠道菌群紊乱和肠屏障功能障碍,益生菌干预可以重塑ACLF大鼠的肠道菌群结构,维护ACLF大鼠的肠屏障功能,促进ACLF大鼠肝脏的修复。
【Abstract】:ObjectiveTo investigate the effect of probiotics intervention on rats with acute-on-chronic liver failure (ACLF) and related mechanism. MethodsA total of 44 male Sprague-Dawley rats were randomly divided into six groups using a random number table, i.e., control group 1 (C1 group with 6 rats without any intervention), model group 1 (M1 group with 8 rats treated with intraperitoneally injected 40% CCl4 oil solution for 10 weeks, followed by phosphate-buffered saline by gavage since week 7 and D-galactosamine acute attack at the end of week 10), probiotics intervention group 1 (Y1 group with 8 rats treated with the same modeling method as the M1 group, and probiotics solution was given by gavage), control group 2 (C2 group with 6 rats without any intervention), model group 2 (M2 group with 8 rats treated with intraperitoneally injected 40% CCl4 oil solution, followed by D-galactosamine acute attack at the end of week 10 and phosphate-buffered saline by gavage at 48 hours after attack, and they were sacrificed at the end of week 12), and probiotics intervention group 2 (Y2 group with 8 rats treated with the same modeling method as the M2 group, and probiotics solution was given by gavage). Body weight, liver function, and liver histopathology were observed before and after intervention. ELISA was used to measure the levels of endotoxin and endocrine immunoglobulin A (sIgA) in plasma, Western Blot was used to measure the content of the intestinal tight junction protein Occludin, RT-PCR was used to measure the mRNA expression of Occludin and ZO-1, and a selective medium was used to measure the changes in related indices including intestinal flora. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the SNK-q test was used for further comparison between two groups. ResultsThere were significant differences in body weight, liver index, alanine aminotransferase, aspartate aminotransferase, total bilirubin, plasma endotoxin, sIgA, and mRNA expression of Occludin and ZO-1 between the C1, M1, and Y1 groups (F=27.65, 8.96, 61.37, 18.27, 21.00, 87.01, 67.10, 101.50, and 10540, all P<0.05), as well as between the C2, M2, and Y2 groups (F=14.04, 12.85, 14.02, 11.39, 35.80, 19.14, 15.37, 2502, and 126.00, all P<0.05). There was no significant difference in the protein expression of Occludin between the C1, M1, and Y1 groups (F=16.40, P<0.05). There were significant differences in the content of Lactobacillus, Bifidobacterium, Enterococcus, and Enterobacter between the C1, M1, and Y1 groups (F=77.95, 66.61, 25.63, and 33.29, all P<0.05), as well as between the C2, M2, and Y2 groups (F=21.50, 22.62, 6.71, and 17.74, all P<0.05). ConclusionIntestinal flora disturbance and intestinal barrier dysfunction are observed in rats with ACLF, and probiotics intervention can remodel the structure of intestinal flora, maintain intestinal barrier function, and promote liver repair.
【关键字】:慢加急性肝功能衰竭; 胃肠道微生物组; 大鼠, Sprague-Dawley; 疾病模型, 动物
【Key words】:acute-on-chronic liver failure;gastrointestinal microbiome; rats, Sprague-Dawley; disease models, animal
【引证本文】:ZHANG YC, BI YZ, FANG X, et al. Protective effect of probiotics in rats with acute-on-chronic liver failure and related mechanism[J]. J Clin Hepatol, 2019, 35(7): 1570-1575. (in Chinese)
张永超,毕研贞,方萧,等. 益生菌对慢加急性肝衰竭大鼠模型的保护作用及其机制[J]. 临床肝胆病杂志, 2019, 35(7): 1570-1575.

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