调肝理脾法对非酒精性脂肪肝病大鼠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 Beclinl in non- alcoholic fatty liver disease in rats

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Beclin-1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（n=10）。模型组、调肝理脾组大鼠皆给予高脂饮食共12周。调肝理脾法组大鼠造模同时给予调肝理牌方2ml／100g／d灌胃，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、肝脏标苓，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Beclinl在肝组织中表达的变化。结果：①HE染色提示模型组大鼠肝细胞可见严重脂肪变性，调肝理睥法组大鼠部分肝细胞可见脂肪变性，其脂肪变性程度低于模型组，差异有统计学意义（P〈0．05），正常对照组大鼠无脂肪变性。②除HDL-C外，模型组大鼠血清AlIrr、AST、TG、TC、LDL—C的水平与正常对照组比较均升高，差异有统计学意义（P〈0．05）；除HDL—C外，调肝理脾法组大鼠血清ALT、AST、TG、TC、LDL-C的水平与模型组比较均降低，差异有统计学意义（P〈0．05）。⑧模型组大鼠肝组织Beclin-1蛋白表达水平与正常对照组比较略增高，但差异无统计学意义（P〉0．05），与模型组比较明显增高，差异有统计学意义（P〈0．05）。结论：调肝理牌法对买验性非酒精性脂肪肝病大鼠的肝功能、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 metabolisnl and the expression of Beclinl in non - alcoholic fatty- liver disease in rats, and to explore the impossible meehanisnl of the method of regulating liver and spleen on non - alco- holic fatty- liver disease. Methods ：30 male healthy SPF SD rats were randonfized into the control group, high - fat group and MRLS group （ 10 nfice 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 intragastriely in the meantinle. At the end of 12th week, all the rats were sac- rificed 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 Beelinl were detected by HE stain, automatic biochemistry analyzers and immunohisto- chemistry respectively. Results： （1）The HE stain showed severe hepatoeellular fatty- degeneration in the high -fat group , and partly hepato-cellular fatty- degeneration in the MRLS group. The degree of fatty- degeneration in the MILLS 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 increased 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 in- creased significantly（ P 〈 0.05 ）. Conclusion： The method of regulating liver and spleen had a good improvement on liver function and blood lipid in rats, and the mechanism was possibly correlated with the increased expression levels of autophagy - associated proteins in liver of rats.