Molecular Mechanism Of Banxia Xiexin Decoction Regulating Oxidative Stress And NF-κB/iNOS Signaling Pathway To Improve T2DM-IR

Objective:To observe the effect of Banxia Xiexin Decoction on oxidative stress and expression of nuclear transcription factor-κB(NF-κb),inducible nitric oxide synthase(iNOS),phosphatidylinositol 3-kinase(PI3K)in rats with type 2 diabetes mellitus(T2DM),and to explore the effect of Banxia Xiexin Decoction on improvement of the molecular mechanism of T2DM-Insulin resistance(T2DM-IR).Methods:120 male specific pathogen free(SPF)Sprague Dawley(SD)rats were randomly divided into the normal control group(10 rats)and T2DM model(110 rats)after one week after the adaptive feeding of the general standard diet.The normal control group was fed with the general diet,while the T2DM group was fed with the high-fat and high sugar diet for 15 days,then fasting for 12 hours,intraperitoneal injection of Streptozotocin(STZ)with the amount of 35 mg/kg,5 days later repeat the same manipulation.Do the whole manipulation for 2 times.72 hours after the completion of the construction model,the tail blood was taken to detect fasting blood glucose.Under the condition of fasting,the tail cutting method was used to select the fasting blood glucose value in the range of 16.7-30.0 mmol/L,and the T2DM model with obvious polydipsia and polyuria was established.The model rats with type2 diabetes were randomly divided into model control group,metformin group,Banxia Xiexin Decoction high-dose group,medium dose group and low dose group.There were 10 rats in each group.During the experiment,the rats in the model group and the intervention group were fed with high-fat and high sugar diet continuously;the rats in metformin group,high,medium and low dose groups of Banxiaxiexin Decoction were fed with 0.16g/kg,13.96g/kg,6.98g/kg and 3.49g/kg respectively.The rats in the normal control group and the model group were fed with the same body volume of normal saline for 12 weeks.During the experiment,the general state of the rats in each group was observed.The amount of diet and drinking water were recorded regularly every week.The changes of body weight and blood glucose were detected.After taking samples,detect the pathological changes of pancreas and liver tissue,blood lipid,liver function,fasting serum insulin,oxidative stress indexes: superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(glutathione peroxidases)The expression of E,GSH Px,catalase,cat,NF-B,iNOS and PI3K.Results:1)basic information of rats: three weeks before gastric administration,the rats in the high,medium and low dose groups and metformin group took more food and drinking water.Their bodies were emaciated.In the process of the experiment,some rats’ hair color gradually appeared withered,yellow and lusterless,more intake food and drinking water,fidgety,easily frightened,and their weight decreased.However,after the third week of administration,the overall situation of the rats in Banxia Xiexin Decoction group and metformin group was significantly improved,their mental state was better,and the glossiness of their fur was better improved.The amount of food and drinking water taken were decreased compared with the model group,but their weight showed an increasing trend.2)fasting blood glucose test results: The high dose group of Banxia Xiexin Decoction,the fasting plasma glucose(FPG)of rats began to decrease at the 3rd week,and the FPG level was stable around 16-20 mmol/L at the 6th-12 th week.The medium dose group of Banxia Xiexin Decoction,the FPG level of rats decreased significantly at the 5th week,and the FPG level was stable around 14-18 mmol/L at the 5th-12 th week.The low dose group of Banxia Xiexin Decoction,the FPG level began to decrease in the 4th week,and the FPG level stabilized around 18-20 mmol/L at the 9th-12 th week.During the experiment,the FPG level in the high,middle and low dose groups of Banxia Xiexin Decoction decreased significantly than the model control group(p<0.05).3)insulin test results: compared with the model control group,the serum insulin levels of the high,middle and low dose groups of Banxia Xiexin Decoction and metformin group were slightly lower,the difference was significant(p<0.05).Compared with the normal control group,the insulin level in the high,middle and low dose groups of Banxia Xiexin Decoction increased,but the difference was not statistically significant.4)insulin resistance level result: After calculating the insulin resistance index,it was found that compared with the model control group,the expression level of insulin resistance index(IRI or HOMA-IR)in the high,middle and low dose groups of Banxia Xiexin Decoction was decreased,the difference was significant(p<0.01).Compared with the normal control group,the expression level of IRI in the high,middle and low dose groups of Banxia Xiexin Decoction was slightly higher,the difference was significant(p<0.01).5)liver function results: Banxia Xiexin Decoction can reduce the activity level of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP).Compared with the model control group,the AST activity level of the high,middle and low dose group of Banxia Xiexin Decoction decreased significantly(p<0.05);the ALT activity level of the high dose group of Banxia Xiexin Decoction decreased significantly(p<0.05);the ALT activity level of the medium and low dose group of Banxia Xiexin Decoction decreased,but the difference was not statistically significant;the ALT activity level of Banxia Xiexin Decoction decreased the activity level of ALP in the low dose group decreased significantly(p<0.05);the activity level of ALP in the high and middle dose groups of Banxia Xiexin Decoction decreased slightly,the difference was not statistically significant.6)blood lipid results: Banxia Xiexin Decoction can reduce the level of total cholesterol(TC)and triglycerides(TG).Compared with the model control group,the level of TC in the high,medium and low dose groups of Banxia Xiexin Decoction decreased,but the level of TC in the middle dose group decreased significantly(p<0.05).TG levels of the Banxia Xiexin Decoction of high,medium and low dose groups were significantly reduced(p<0.05).Banxia Xiexin Decoction can reduce serum low-density lipoprotein cholesterol(LDL-C)and increase the level of high-density lipoprotein cholesterol(HDL-C).Compared with the model control group,the HDL-C level of Banxia Xiexin Decoction of high,medium and low dose groups is slightly higher,the difference is not statistically significant.The LDL-C level of all rats in the Banxia Xiexin Decoction of high,medium and low dose groups decreased,but the difference was not statistically significant.7)Oxidative stress index of pancreatic tissue results: Banxia Xiexin Decoction can reduce MDA content and increase CAT,GSH-Px and SOD activity levels.Compared with the model control group,the level of MDA in the high dose group of Banxia Xiexin Decoction decreased,but the difference was not statistically significant;the level of CAT activity in the medium dose group of Banxia Xiexin Decoction was slightly higher,but the difference was not statistically significant;the level of GSH PX activity in the medium dose group of Banxia Xiexin Decoction increased significantly(p<0.01);GSH-Px in the medium dose group of Banxia Xiexin Decoction increased,there was no significant difference;The expression level of SOD activity in the high and medium dose group of Banxia Xiexin Decoction,the difference was significant(p<0.05);The expression level of SOD activity in the low dose group of Banxia Xiexin Decoction,but there was no statistical significance.8)Liver oxidative stress index results: Banxia Xiexin Decoction can reduce MDA content and increase GSH-Px and SOD activity level.Compared with the model control group,the MDA content in the high,medium and low dose groups of Banxia Xiexin Decoction decreased significantly(p<0.05);the GSH-Px activity expression level in the medium and low dose groups of Banxia Xiexin Decoction increased,but the difference was not statistically significant;The SOD activity expression level in the high dose group of Banxia Xiexin Decoction increased significantly(p<0.01).The level of SOD activity in the medium dose group of Banxia Xiexin Decoction also increased,but the difference was not statistically significant.9)PI3K protein in liver results: Compared with the model control group,the expression of PI3K protein in the high and low dose group of Banxia Xiexin Decoction was significantly higher,while the expression of PI3K protein in the medium dose group of Banxia Xiexin Decoction was lower.Compared with the model control group,the expression level of PI3K in the high and medium dose groups of Banxia Xiexin Decoction increased,but the difference was not statistically significant.10)NF-κB protein in liver results: Banxia Xiexin Decoction can reduce the level of NF-κB.Compared with the model control group,the expression of NF-κB protein in the high and medium dose groups of Banxia Xiexin Decoction was significantly reduced,while the expression of NF-κB protein in the low dose group of Banxia Xiexin Decoction was decreased in varying degrees.Compared with the model control group,the expression level of NF-κB in the high and medium dose group of Banxia Xiexin Decoction was significantly decreased(p<0.05);the expression level of NF-κB in the low dose group of Banxia Xiexin Decoction was lower,but the difference was not statistically significant.11)iNOS protein in liver results: Banxia Xiexin Decoction can reduce the expression level of iNOS.Compared with the model control group,the expression of iNOS protein in the high,medium and low dose groups of Banxia Xiexin Decoction decreased.Compared with the model control group,the expression level of iNOS in the high,medium and low dose groups of Banxia Xiexin Decoction was significantly decreased(p<0.05).12)pathological test results: Compared with the histopathology of the pancreas and liver of the rats,it was found that in the model control group,a small number of islet cells were necrotic,and a small number of islet cells were loose and vacuolized;Inflammatory cell infiltration was found in the liver,and a small number of hepatocyte steatosis,mild edema,cell swelling were found;no other abnomal cells were found in the pancreas and liver Obviously abnormal.The pathological structure of pancreas and liver in metformin group,high,medium and low dose of Banxia Xiexin Decoction group were improved,and no other obvious abnormality was found.Conclusions:1)Banxia Xiexin Decoction can correct the disorder of glucose,lipid metabolism and improve IR in T2DM rats to a certain extent.2)Banxia Xiexin Decoction can increase SOD and GSH-Px in pancreas and liver tissue of T2DM rats,the MDA expression level reduced,which indicates that the improvement of T2DM-IR may be related to the regulation of oxidative stress.3)Banxia Xiexin Decoction can regulate the expression of NF-κB,iNOS and PI-3K protein in the liver of T2DM rats,and the regulation of PI-3K protein related to insulin signal transduction may be related to the improvement of T2Dm-IR by Banxia Xiexin Decoction.