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调肝理脾法对非酒精性脂肪性肝病大鼠模型脂质代谢及微管相关蛋白轻链3B表达的影响
Effect of liver-regulating and spleen-strengthening method on lipid metabolism and expression of microtubule-associated protein light chain 3B in a rat model of nonalcoholic fatty liver disease
文章发布日期:2018年03月07日  来源:  作者:李长新,周滔,牛柯敏,等  点击次数:421次  下载次数:53次

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【摘要】:目的观察调肝理脾法对大鼠非酒精性脂肪性肝病(NAFLD)的改善作用及对大鼠肝组织微管相关蛋白轻链3B(LC3B)表达的影响,探讨调肝理脾法治疗NAFLD的作用机制。方法SD雄性大鼠(清洁级)30只,随机分为对照组、模型组、调肝理脾法组,每组10只。模型组、调肝理脾法组大鼠皆予以高脂饮食,共12周。调肝理脾法组大鼠造模同时给予调肝理脾方2 ml·100 g-1·d-1灌胃(相当于成人剂量的5倍),对照组大鼠给予普通饲料及等量生理盐水灌胃。第12周末处死动物,采集血液、肝脏标本,检测各组大鼠血清ALT、AST、TG、TC、HDL-C、LDL-C水平,并通过HE染色观察肝脏脂肪变性情况,免疫组化法检测LC3B在肝脏组织中表达的变化。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LSD-t检验;计数资料组间比较采用χ2 检验;等级资料多组间比较采用Kruskal-Wallis H检验,进一步两两比较采用Mann-Whitney U检验。结果 HE染色提示模型组大鼠肝细胞脂肪变性程度和炎症活动度计分与正常对照组相比,差异均有统计学意义(P值均<0.001);调肝理脾法组脂肪变性程度和炎症活动度计分明显减轻,与模型组相比差异均有统计学意义(P值均<0.01)。与正常对照组比较,模型组大鼠血清ALT、AST、TG、TC、HDL-C、LDL-C的水平均升高,差异均有统计学意义(P值均<0.001)。与模型组比较,除HDL-C外,调肝理脾法组大鼠血清ALT、AST、TG、TC、LDL-C的水平均降低,差异均有统计学意义(P值均<0.05)。与正常对照组比较,模型组大鼠肝组织LC3B表达水平略增高,差异无统计学意义(χ2 =1.250,P>0.05);与模型组比较,调肝理脾法组肝组织LC3B表达水平明显增高,差异有统计学意义(χ2 =5.051,P<0.05)。结论调肝理脾法对实验性SD雄性NAFLD大鼠肝功能、血脂等有较好的改善作用,其作用机制可能与升高SD雄性大鼠肝组织内的自噬水平有关。
【Abstract】:ObjectiveTo investigate the effect of the liver-regulating and spleen-strengthening method on lipid metabolism and expression of microtubule-associated protein light chain 3B (LC3B) in a rat model of nonalcoholic fatty liver disease (NAFLD), as well as the mechanism of this method in the treatment of NAFLD. MethodsA total of 30 clean male Sprague-Dawley rats were randomly divided into control group, model group, and liver-regulating and spleen-strengthening group, with 10 rats in each group. The rats in the model group and the liver-regulating and spleen-strengthening group were given high-fat diet for 12 weeks. The rats in the liver-regulating and spleen-strengthening group were given liver-regulating and spleen-strengthening prescription 2 ml·100 g-1·d (five times the dose for adults) by gavage, and those in the control group were given normal diet and an equal volume of normal saline by gavage. The rats were sacrificed at the end of week 12, and blood and liver samples were collected to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). HE staining was performed to observe hepatic steatosis, and immunohistochemistry was used to measure the change in the expression of LC3B in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between groups; the Kruskal-Wallis H test was used for comparison of ranked data between multiple groups, and the Mann-Whitney U test was used for further comparison between two groups. ResultsHE staining showed that there were significant differences in the degree of hepatocyte fatty degeneration and score of inflammatory activity between the model group and the control group (both P<0.001); compared with the model group, the liver-regulating and spleen-strengthening group had significant reductions in the degree of hepatocyte fatty degeneration and score of inflammatory activity (both P<0.01). Compared with the control group, the model group had significant increases in the serum levels of ALT, AST, TG, TC, HDL-C, and LDL-C (all P<0.001); compared with the model group, the liver-regulating and spleen-strengthening group had significant reductions in the serum levels of ALT, AST, TG, TC, and LDL-C (all P<0.05). Compared with the control group, the model group had a slight increase in the expression of LC3B in liver tissue (χ2=1.250, P>0.05); compared with the model group, the liver-regulating and spleen-strengthening group had a significant increase in the expression of LC3B in liver tissue (χ2=5.051, P<0.05). Conclusion The liver-regulating and spleen-strengthening method can improve liver function and blood lipids in experimental male Sprague-Dawley rats with NAFLD, possibly by increasing the level of autophagy in liver tissue.
【关键字】:调肝理脾法; 非酒精性脂肪性肝病; 自噬; 微管相关蛋白质类; 大鼠, Sprague-Dawley
【Key words】:method of regulating liver and spleen; non-alcoholic fatty liver disease; autophagy; microtubule-associated proteins; rats, Sprague-Dawley
【引证本文】:LI CX, ZHOU T, NIU KM, et al. Effect of liver-regulating and spleen-strengthening method on lipid metabolism and expression of microtubule-associated protein light chain 3B in a rat model of nonalcoholic fatty liver disease [J]. J Clin Hepatol, 2018, 34(4): 829-835. (in Chinese)
李长新, 周滔, 牛柯敏, 等. 调肝理脾法对非酒精性脂肪性肝病大鼠模型脂质代谢及微管相关蛋白轻链3B表达的影响[J]. 临床肝胆病杂志, 2018, 34(4): 829-835.

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