Study On The Mechanism Of Gexia Zhuyu Decoction In The Treatment Of NAFLD Based On Autophagy

ObjectiveTo investigate the effect of Gexia zhuyu decoction in the treatment of NAFLD from perspective of autophagy.Whether the effect is related to the expression of autophagy-related proteins in live,to reveal the mechanism of treatment,and provide theoretical basis for the clinical application and experimental research of this prescription.Methods1.The SD male rats were divided into 6 groups randomly: Control group,Model group,Gexia Zhuyu Decoction’s low-dose group,Gexia Zhuyu Decoction’s medium-dose group,Gexia Zhuyu Decoction’s high-dose group,Rapamycin group.After 6 weeks,the model was established,the low,medium,high-dose group of Gexia Zhuyu Decoction were feded with 7.2g/kg/d,14.4g/kg/d and 28.8g/kg/d of Gexia Zhuyu Decoction respectively;Rapamycin group was feded with 2.8mg/kg/d of rapamycin;the control group and the model group were feded with 2 m L /d of normal saline.At the 8th,10 th,12th week,8 animals were collected from each group.2.To observe the therapeutic effect of Gexia Zhuyu Decoction on NAFLD rats.At the8 th,10th and 12 th week,pathological changes of liver tissue were observed by HE staining of paraffin section and oil red O staining of frozen section.Liver function indexes of Serum(AST,ALT),blood lipid of rats(TC,TG,LDL-C,HDL-C)were detected by colorimetry.Liver fat(TC,TG)of rats was detected by GPO-PAP method.3.To observe the effect of Gexia Zhuyu Decoction on the expression of autophagy related proteins in NAFLD rats.The protein expressions of Beclin1,LC3B-Ⅱ and p62 in liver tissues and the ratio of LC3B-Ⅱ/LC3B-Ⅰ were detected by Western Blot.Immunohistochemistry(IHC)was used to detect the protein expressions of p62 and ATG5 in liver tissues.Result1.After 6 weeks of feeding with high-sugar and high-fat diet,NAFLD rats were modeled successfully.The model has the characteristics: the body weight and liver index of NAFLD rats increased significantly;There were different size fat vacuoles in liver cells of NAFLD rats;The serum TC,TG,AST,ALT,LDL-C and hepatic lipid TG and TC of NAFLD rats were increased significantly,while the HDL-C was decreased significantly.The protein expressions of Beclin1,LC3B-Ⅱ,Atg5 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ in NAFLD rats were decreased significantly,while the protein expression of p62 was increased significantly.2.After NAFLD rats were modeled successfully,Gexia Zhuyu Decoction was used to treat NAFLD rats.Gexia Zhuyu Decoction can reduce the liver index of NAFLD rats effectively.To reduce the degree of steatosis in liver tissue of NAFLD rats;The serum levels of TC,TG,AST,ALT,LDL-C and hepatic lipid TG and TC of NAFLD rats were decreased,and the serum HDL-C level was increased;The protein expression of Beclin1,LC3B-Ⅱ,Atg5 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ in NAFLD rats were enhanced,and the protein expression of p62 was decreased.Some of these indexes improved more obviously with the prolongation of treatment time.3.After NAFLD rats were modeled successfully,NAFLD rats were treated with rapamycin.During the whole course of rapamycin treatment,serum TC,TG,LDL-C and HDL-C of NAFLD rats were increased significantly;The serum AST,ALT,and hepatic lipid TC,TG of NAFLD rats were decreased at the end of 8th and 10 th week,reduce the degree of steatosis in liver tissue of NAFLD rats at the end of 8th and 10 th week,and the indexes were increased at the end of 12 th week;The protein expression of Beclin1,LC3B-Ⅱ,ATG5 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ in NAFLD rats were enhanced,and the protein expression of p62 was decreased.Conclusion1.Gexia Zhuyu Decoction can reduce the steatosis degree of liver tissue of NAFLD rats,and it can improve the liver index,serum TC,TG,AST,ALT,LDL-C,HDL-C and TC and TG in the liver fat of NAFLD rats effectively,so it has a good effect on the treatment of NAFLD.2.Gexia Zhuyu Decoction can improve the expression of autophagy-related proteins Beclin1,LC3B-Ⅱ,P62,ATG5 in liver tissue and the ratio of LC3B-Ⅱ/LC3B-Ⅰ of NAFLD rats effectively,the mechanism of action may be related to the enhancement of autophagy level in liver tissue of NAFLD rats.