| Objective: Qutanhuoxue decoction is an experience prescription of Professor Sun Tongjiao in our hospital in the treatment of non-alcoholic fatty liver disease.Preliminary research have shown that Qutanhuoxue decoction can significantly improve sputum-stasis-interstitial non-alcoholic steatohepatitis,significantly reduce the degree of fatty liver changes in liver tissue of non-alcoholic fatty liver disease rats,reduce liver inflammation,and regulate liver lipid metabolism,the mechanism of action is not fully understood.Insulin resistance is the central link of non-alcoholic fatty liver disease,using the rat model of non-alcoholic fatty liver disease to explore the mechanism of Qutanhuoxue decoction regulating the important signal transduction pathway of insulin IRS/PI3K/AKT/mTOR signal pathway in the treatment of non-alcoholic fatty liver disease,further clarify the molecular mechanism of Qutanhuoxue decoction in the treatment of non-alcoholic fatty liver disease,and provide an experimental basis for the clinical experience and academic thought of famous old Chinese medicine.Method: 1.Grouping,model building and administration(1)After 30 SD rats were adaptively fed for 1 week,use random number table to randomly into normal group(10)and build module group(20);the normal group was fed with ordinary feed,and the build module group was fed with high-fat feed.Normal activities and drinking water of rats in each group.(2)The modeling time was 8 weeks.After 8 weeks,2 model rats were randomly selected and sacrificed.Take the left lobe of the liver and fix it in neutral formalin,and perform HE staining.Based on the results of liver pathology testing and NAS scoring system,it was determined whether the model standards were met.The remaining build module group(18 animals)was randomly divided into model group(9 animals)and Qutanhuoxue decoction group(9 animals).(3)The normal group continued to be fed with ordinary feed,while the model group and the expectorant and blood circulation group were fed with high-fat feed.The normal group and the model group were intragastrically administered with 10 ml /(kg·d)normal saline,and the expectorant and blood circulation formula group was administered with the antiphlegm and activating blood circulation extract 11.7g /(kg·d)intragastrically,after 4 weeks of continuous intervention.2.Specimen collection and processing(1)On the 4th week of intervention,the rats were weighed and anesthetized by intraperitoneal injection of 10% chloral hydrate(3 mL/ kg).The abdominal aorta was separated and exposed.The blood volume was about 5 mL.After standing for 4 hours,centrifuged for 10 min to obtain the upper serum-Store at-80 ?.(2)The rats were sacrificed,and the livers were removed and weighed.After washing with physiological saline,the paper was blotted with filter paper and weighed.Take the left lobe of the liver and fix it in 4% paraformaldehyde,and perform HE staining;another section of the right lobe of the liver,about 1.0cm×1.0cm×1.0cm in size,was frozen and stained with oil red 0;the remaining liver was put into frozen storage Store in tube at-80 ? 3.To observe the effects of Qutanhuoxue decoction on serum biochemical parameters in NAFLD rats(1)The kit detects serum TG,TC,LDL-C,HDL-C,FBG content(2)Detection of FINS and VLDL by ELISA(3)Calculating HOMA-IR 4.Observe and explore the effect of Qutanhuoxue decoction on IRS/PI3K/AKT/mTOR signaling pathway Expression of IRS/PI3K/AKT/mTOR signaling pathway related proteins by Qutanhuoxue recipe by western blotting.5.To observe the effect of Qutanhuoxue decoction on liver pathological changes in NAFLD rats Changes of liver pathology in NAFLD rats by HE and oil red O staining Result: 1.Qutanhuoxue decoction can reduce weight and wet liver weight in rats with non-alcoholic fatty liver disease.2.Qutanhuoxue decoction can significantly reduce TG,TC,LDL-C,VLDL,increase HDL-C,significantly reduce FBG,FINS,HOMA-IR,regulate glucose and lipid metabolism,and improve insulin resistance in NAFLD rats.3.Qutanhuoxue decoction can significantly improve the liver lipid deposition and steatosis in NAFLD rats.4.Qutanhuoxue decoction can reduce the expression of IRS-1,p-PI3 K,p-AKT and mTORC1 proteins in liver tissue of NAFLD rat model Conclusion: 1.Qutanhuoxue decoction can improve insulin resistance in NAFLD rats,regulate glucose and lipid metabolism,and improve the degree of liver fatty changes in NAFLD rats.2.The mechanism of Qutanhuoxue decoction in the treatment of NAFLD may be to play a role by inhibiting the expression of genes and proteins related to the IRS-PI3K-AKT-mTOR signaling pathway. |
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