调肝理脾法对大鼠非酒精性脂肪性肝病脂质代谢和自噬相关蛋白-1表达的影响  

Effect of method of regulating liver and spleen on lipid metabolism and expression of Beclin1 in non-alcoholic fatty liver disease in rats

作　　者：李长新 周滔 牛柯敏[2,3] 陈誩 危北海[2] 王佳[1] 孟思宏 李沁娜 郑娇 申青艳 陈瑞琳 LI Chang-xin;ZHOU Tao;NIU Ke-min(Department of Pathology,Chinese Medical Hospital of Shunyi District（Beijing,101300）China.)

机构地区：[1]北京中医医院顺义医院病理科,北京顺义131000 [2]首都医科大学附属北京中医医院消化中心 [3]北京中医药大学北京中医临床医学院

出　　处：《中西医结合肝病杂志》2018年第5期290-293,322,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：北京市卫生系统高层次卫生技术人才培养计划(No:2015-3-114);北京市科委“首都临床特色应用研究”专项课题(No:Z141107002514082);北京中医医院-北京中医医院顺义医院科研合作基金项目(No:SYSY-201410)

摘　　要：目的:观察调肝理脾法(MRLS)对实验性大鼠非酒精性脂肪肝病(NAFLD)的改善作用及对大鼠肝组织自噬相关蛋白Beclin1表达的影响,探讨调肝理脾法治疗非酒精性脂肪肝病的可能作用机制。方法:SD雄性大鼠(清洁级) 30只,随机分为对照组、模型组、调肝理脾组(各10只)。模型组、调肝理脾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2ml/100g/d灌胃(相当于成人剂量的5倍),对照组大鼠给予正常饮食(普通饲料)及等量生理盐水灌胃。第12周末处死动物后,分别采集大鼠血液、肝脏标本,检测各组大鼠血清ALT、AST、TG、TC、HDL-C、LDL-C水平,通过HE染色观察肝脏脂肪变性情况,免疫组化法检测Beclin1在肝组织中表达的变化。结果:(1)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,差异有统计学意义(P <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,与模型组相比差异有统计学意义(P <0. 01)。(2)与对照组比较,模型组大鼠血清ALT、AST、TG、TC、HDL-C、LDL-C的水平均升高,差异有统计学意义(P <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,差异有统计学意义(P <0.05)。(3)与对照组比较,模型组大鼠肝组织Beclin1蛋白表达水平略增高,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Beclin1蛋白表达水平明显增高,差异有统计学意义(χ2=6. 121,P <0. 05)。结论:调肝理脾法对实验性大鼠非酒精性脂肪肝病的肝功能、血脂等指标有很好的改善作用,其作用机制可能与升高大鼠肝组织内的自噬水平有关。Objective： To observe the effect of method of regulating liver and spleen on lipid metabolism and the expression of Beclinl in non-alcoholic fatty liver disease in rats, and to explore the impossible mechanism. Methods：Thirty male healthy SPF SD rats were randomized into the control group,high-fat group and MRLS group （ 10 mice in each group） ; the rats of high-fat group and MRLS group were fed with high fat diets for 12 weeks , and the MRLS group being fed with high-fat diet was applied by the method of regulating liver and spleen（2ml/ 100g/d） at the same time. The rats in control group was fed with standard diets and saline of the same volume was perfused intragastricly in the meantime. At the end of 12th week, all the rats were sacrificed and we collected the blood samples and liver tissues from the rats immediately. The fatty degeneration , the serum levels of ALT, AST,TG,TC, HDL-C, LDL-C, , and the expression of Beclinl were detected by HE stain, automatic biochemistry analyzers and immunohistochemistry respectively. Results： （1）The HE stain showed severe hepatocellular fatty degeneration in the high-fat group , and partly hepatocellular fatty degeneration in the MRLS group. The degree of fatty degeneration in the MRLS group was significantly lower than that in the high-fat group （P 〈 0.05 ）. There was no fatty degeneration in the control group. （2）Compared with those of the control group, beyond HDL-C increased slightly,the serum ALT,AST,TG,TC, LDL-C levels of the high-fat group in- creased significantly（P 〈 0.05） ; Compared with those of the high-fat group, beyond HDL-C increased slightly, the serum ALT, AST, TG, TC, LDL-C levels of the MRLS group reduced significantly（ P 〈 0.05 ）. （3）Compared with those of the control group, the expression of Beclinl in high-fat group increased slightly, and there was no significant difference （P 〉 0.05） ; Compared with those of the high-fat group, the expression of Beclinl in the MRLS group increased significantly（P 〈

关 键 词：调肝理脾法 大鼠 非酒精性脂肪性肝病 自噬 BECLINL 

分 类 号：R285.5[医药卫生—中药学]