The Influence And Its Mechanism Of Banxia Xiexin Decoction Drug Serum On The RIN-m5F Pancreas Islet Cell Apoptosis

T2DM is one of the chronic non-communicable diseases which has genetic predisposition and can be influenced by environmental and other factors.Pancreas islet ? cell apoptosis is one of the key pathogeneses of T2DM.So how to reduce P cells apoptosis become one of the important methods for the prevention and control of diabetes and its complications.Many Chinese medicine scholars think Pi Xu Wei Re,Tan Yu Hu Jie,gastrointestinal disorder is one of the important pathogeneses which can lead to the development of diabetes.The harmonic gastrointestinal method of traditional Chinese medicine can improve the function of the gastrointestinal,invigorate spleen and clear heat,promote blood circulation and eliminate sputum,so it can be used for the treatment of diabetes and its related diseases.Part One Literature ReviewThe incidence of T2DM showed a trend of rising in recent years.It can cause serious impact on people's life quality and life safety,and bring huge economic burden.T2DM can be caused by a variety of reasons,such as genetic factor and environ-mental factor.The current research on T2DM Etiology more focused on thePancreas islet ? cell apoptosis.More and more studies confirmed that the glucolipotoxicity,high concentration of free fatty acids and a variety of factors such as inflammatory cytokines can increase ? cell oxidative stress which canlead to ?cells apoptosis,induce or aggravate the course of T2DM.With the development of further research about the cause of diabetes,the treatment mechanism of many drugs are being researched,such as GLP-1 analogues and DPP-IVinhibitors,many studies have shown that these drugs can inhibit the apoptosis of pancreatic islet ? cells and promote the activity of GLP-1 which is helpful for the improvement of T2DM.But there are many disadvantages about these drugs,such as not included in the health care,expensive,short comings of preserving and inconvenient to carry.Many Chinese medicine scholars think Pi Xu Wei Re,Tan Yu Hu Jie,gastrointestinal disorder is one of the important pathogenesiseswhich can lead to the development of diabetes.The harmonic gastrointestinal method of traditional Chinese medicine can improve the function of the gastrointestinal,invigorat spleen and clear heat,promote blood circulation and eliminate sputum.And the foothold of banxia xiexin decoction is gastrointestinal regulation which coincides with the "gastric-bowel-pancreas islet"axis.So it can be used for the treatment of diabetes and its related diseases.Modern clinical and experimental research suggests that this decoction and its monomer can improve glucose and lipid metabolism,reduce blood sugar,improve the gastrointestinal hormone secretion and body's antioxidant ability and immune function,and inhibit the apoptosis of pancreatic islet cells.But the mechanism of this decoction is unclear,itneeds to be tested and verified.Part Two Experimentatal ResearchObject:We cultured the RIN-m5F cell in vitro,and then intervened with drug.The object was confirming the decoction can restain cell apoptosis and explaining its mechanism and proving the scientificity of the decoction's treatment of T2DM through the methods of CCK8,AV/PI and Western blot.Method:33 male SD rats were randomly assigned to be gavaged for seven days,then drug-containing serums were collected(normal serum,Banxiaxiexin-containing serum,Liraglutide-containing serum).H202 was used to establish apoptosis model and the optimum concentration of the decoction was explored.Cells were randomly assigned to four groups:normal control group,model group,H202+BXXXTgroup,H202+GLP-1 groups.AV/PI was used to detect apoptosis rate,RIA was used to measure the quantity of insulin,and Western blot was used to determine the protein expression of Bcl-2,Bax,Caspase 3,PI3K,AKT and p-AKT.The data management and processing were employing Microsoft EXCEL 2010 and SPSS20.0,with meant standard deviation(x ±s)express.Adopting the method of One-Way ANOVA after normality and homogeneity of variance test,and the LSD-t test was used to process intergroup comparison with statistical significance being P<0.05.Results:(1)The apoptosis model was established.The suitable concentration of H202 is 300?m.(2)The optimal concentration of banxiaxiexin decoction was established.The protection of different concentration of banxiaxiexin decoction on islet cell was tested by CCK8.It was found that the best concentration was 10%,the difference had statistical significance(P<0.05).(3)The protection function was confirmed.The flow cytometry results showed that compared with model group,the cell vitality was inhanced remarkably and the apoptosis rate was decreased signally after drug intervention,and the difference had statistical significance(P<0.05).(4)We confirmed that the this decoction can promote the secretion of insulin.The insulin secretion was detected by RIA.The result showed that compared with model group,the basic and glucose stimulated insulin secretion were significantly increased after drug intervention,and the difference had statistical significance(P<0.05).(5)We verified the decoction could down-regulate the expression of Bax,Caspase-3 and up-regulate the expression of Bcl-2 and p-AKT.The related protein expression was tested by Western blot which showed that compared with model group,the expression of Caspase-3 and Bax were down-regulated significantly after drug intervention and the expression of Bcl-2 and p-AKT was up-regulated significantly after drug intervention,and the difference all had statistical significance(P<0.05).Conclusion:Banxiaxiexin decoction could improve pancreas islet ? cells apoptosis and promote the secretion of insulin,which may be related to the activation of PI3K/AKT pathway and then the regulation of downstream factor expression.