| Purpose: In this experiment,investigated the effect of RFQG on the physiological status and blood glucose,oxidative stress indexes and related proteins of type 2 diabetic rats throug regulating Keap1-Nrf2/ARE oxidative stress signaling pathway to prove the therapeutic efficacy and prospect of Chao Yao RFQG on type 2 diabetes mellitus.Methods:1.Preparation of RFQG: medicinal materials were soaked for 2.5 h,steamed and filtered twice,and made into lyophilized powder after spin-steaming and concentration.2.Experimental animal preparation: 8 of 48 SD rats were taken out as the blank group,and the remaining 40 SD rats were fed with high-fat and high-sugar diet for 4 weeks and then fasted for 12 h.30 mg/kg-BW streptozotocin(STZ)was injected by intraperitoneal injection,and fasting blood glucose was measured after 72 h that blood was taken from the severed tail,if the figure of the fasting blood glucose is more than 11.1 mmol/L,it is concerned to be a successful establishment of the diabetic rat model.3.The 30 rats which were successful modeling were randomly divided into the model group,the high,medium and low dose groups of RFQG and the positive group,with 6 rats in each group.The rats were fed with distilled water in the model group,400 mg/kg-BW of RFQG in the high-dose group,200 mg/kg-BW of RFQG in the medium-dose group,100mg/kg-BW of RFQG in the low-dose group,and 200 mg/kg-BW of metformin hydrochloride in the positive group for 6 weeks.4.Measurements: During the experiment,measured the body weight and after 8 hours of fasting blood glucose(FBG)values once a week.At the end of feeding,the glucose tolerance level(Oral Glucose Test(OGTT)was measured,blood was taken from the abdominal aorta of rats to measure triglyceride(TG),total cholesterol(TC),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C)and Malondialdehyde(MDA),Superoxide Dismutase(SOD),Catelase(CAT),Glutataione(GSH)oxidative stress related indexes.Liver tissues were taken and the pathological changes of liver were determined by HE-staining method.The expression of retinal ECH-associated protein1(Keap1),heme oxygenase-1(HO-1),nuclear factor erythroid-2(Nrf2),and quinone oxidoreductase NADH1(NQO1)proteins were measured by Western Blot.Results:1.After the treatment of RFQG,the weight decreased slowly in the high-dose group compared with the model group,and there was a significant difference(P<0.05);RFQG had a significant hypoglycemic effect,and FBG decreased extremely significantly in the high-dose group(P<0.01).2.OGTT levels decreased extremely significantly in the high-dose group compared with the model group(P<0.01).3.The high-dose group compared with the model group,TG,TC,and LDL-C levels were significantly decreased(P<0.01)and HDL-C levels were significantly increased(P<0.01).4.RFQG could effectively alleviate oxidative stress,MDA levels were significantly decreased(P<0.05)and SOD,GSH,CAT levels were significantly increased(P<0.01)in the high-dose group compared with the model group.5.The HE-staining results showed that in the model group,the hepatocyte cords were disordered,the cells produced fusion with unclear borders,and the cells had different morphology,and a large number of inflammatory cell infiltrates were seen in the hepatic portal canal area,and many vacuolated degeneration existed in the cytoplasm of hepatocytes;in the high-dose group and the positive group,the hepatocyte cords were arranged in an orderly manner,some cells had fusion,but the borders were generally clear,and there were a few inflammatory cell infiltrates in the hepatic portal canal area.6.Western Blot analysis,compared with the blank group,Keap1 protein expression was significantly up-regulated(P<0.05)and NQO1,HO-1 and Nrf2 protein expression was highly significantly down-regulated(P<0.01)in the model group;compared with the model group,Keap1 protein expression was highly significantly down-regulated(P<0.01)in the mid-dose group;NQO1 protein expression was highly significantly up-regulated(P<0.01)and NQO1 protein expression level was significantly up-regulated(P<0.05)in the mid-dose group;HO-1 protein expression was highly significantly up-regulated(P<0.01)and HO-1protein expression was significantly up-regulated(P<0.05)in the high-dose group;Nrf2protein expression level was highly significantly up-regulated(P<0.01)in the mid-dose group.Conclusion: The addition of RFQG soup can effectively regulate body weight,FBG,OGTT and blood lipids in type 2 diabetic rats by modulating the Keap1-Nrf2/ARE oxidative stress signaling pathway,improve the level of oxidative stress and reduce liver injury in type2 diabetic rats,which has significant efficacy in the treatment of type 2 diabetes. |
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