| ObjectiveChronic stress is an important cause of Type 2 diabetes(T2DM),and Insulin resistance(IR)is one of the main pathogeneses of T2 DM.From the perspective of T2 DM in TCM etiology,this study points out that the basic pathogenesis of diabetes is that liver depression and spleen deficiency is the hypothesis about the basic pathogenesis of diabetes development.This study carries out the animal in vivo and in vitro experiments,using modern experimental technology such as pathology,molecular biology,serum pharmacology etc.,and according to Glyco-lipid metabolism and its related gene and protein changes,the study also investigates the mechanism of psychological stress on IR in T2 DM mice and mechanism of Chinese medicine compound and Modified Xiaoyao Powder,which explores a new way to prevent and treat diabetes with TCM.Methods1.In Animal experiment,T2 DM rats were established by streptozotocin(STZ)combined with high glucose and high fat diet.The model of T2 DM rats were randomly divided into four groups: model control group,Xiaoyao San group,stress control group,stress with Xiaoyao San group.The model of chronic psychological stress was established for the latter two groups.At the same time,the treatment group was given Xiaoyao Powder Gavage The blood,liver,pancreas and right gastrocnemius muscle was sacrificed 22 days after the model established.The expressions of GLUT4,IRS-1 and Grp78 were detected by immunohistochemistry and Western Blotting.The levels of HbA1 c,FINS,ISI,TC,TG and LDL-C in each group were measured by ELISA.2.In cell experiments,RIN-m5 f cells were treated with 50 mM glucose to induce a cell model of high glucose-induced injury.Based on this,250 uM corticosterone(CORT)was added to establish a RIN-m5 f cell model of stress-induced diabetes.The established cells were divided into 7 groups: Normal group,G group,Gco group,GC group,GX group,GCoC group and GCoX group.The above groups were intervened with 30% rat serum.The cell viability,insulin level,ATF6 and IRE1 mRNA,Caspase-12 and CHOP protein levels were detected by ELISA,Western-blot and qPCR.Results1.The results of Animal experiments in vivo show that chronic stress stimulation significantly increases FBG,HbA1 c,TG and TCHO and significantly decreases BW,HDL-C and ISI of both T2 DM models,and Modified Xiaoyao powder could reverse the changes of FBG,TG,TCHO,LDL-C,HDL-C,HbA1 c and ISI caused by chronic stress.In terms of protein expression,chronic stress can increase the expression of GLUT4 and IRS-1 in the pancreas,liver and muscle tissue of T2 DM model rats.Xiaoyao powder can up-regulate the expression of GLUU4 and IRS-1 in T2 DM rats,while Modified Xiaoyao powder can reduce the expression of GLUU4 and IRS-1 under chronic stress.2.Vitro cell culture experiments shows that CORT induces islet cell apoptosis under high glucose condition.High glucose,CORT Modified Xiaoyao powder are all factors that promote the secretion of INS.High glucose can down-regulate protein expression of ATF6 and IRE1 mRNA,Caspase-12 and CHOP,while high glucose can up-regulate the expression of ATF6,IRE1 mRNA,Caspase-12 and CHOP in high glucose environment.Besides,Modified Xiaoyao powder can reverse the expression changes of IRE1 mRNA and Caspase-12 induced by CORT.ConclusionChronic psychological stress can aggravate T2 DM model rats,reduce their insulin sensitivity,and increase TG,CHO and reduce HDL-C in their serum so that model rats lose their weight.Modified Xiaoyao powder not only has a better curative effect on T2 DM,but also can alleviate the condition of hyperglycemia and insulin sensitivity of T2 DM model rats under chronic stress,reducing the levels of TG,CHO and LDL-C and increasing the level of HDL-C.Its mechanism may be through regulation of various T2DM-related gene and protein expression,and improve insulin sensitivity in order to correct the model rats of glucose and lipid metabolism disorder. |
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