| ObjectiveTo evaluate the anti-diabetic effects of Modified Banxia Xiexin Decoction by investigating blood glucose,blood lipid,Tumor necrosis factorα(TNF-α)and insulin resistance in Type-2 Diabetes Mellitus(T2DM)rats.To further study the mechanism of hypoglycemic effect through the protein expression of PERK-CHOP signal pathway on pancreatic tissue ERS in T2DM rats,thus providing experimental proofs for treating diabetic with Modified Banxia Xiexin Decoction and a new ideas for clinical treatment of type 2 diabetes mellitus.Method(1)80 SPF grade(140±20)g male Wistar rats after 1 week adopted feed,20 rats were selected randomly as blank group,and feed 10 weeks by ordinary feed.60 rats were treated with high-fat and high-sugar and then intraperitoneal injected streptozotocin(STZ)to establish the model of T2DM rats.The rats with the symptom of polydipsia,polyuria,emaciation and 11.1mmol· L-1≤FBG≤33.3mmol · L-1 were T2DM model rats.Then randomized them into 3 groups,model group,Modified Banxia Xiexin Decoction group and metformin group.After that the rats were administrated with Modified Banxia Xiexin decoction(20g·kg-1,respectively)or metformin(200 mg·kg-1,respectively).Model group and control group animal were administrated with distilled water.All the groups were feed by ordinary feed while therapy.(2)Fasting blood glucose,body weight were recorded at the same time each week.5 weeks later,anesthetize the rats by abdominal injection wit h 2%pentobarbital sodium,and then take the blood serum samples from abd ominal artery,for the detection of fasting blood-glucose(FBG),fasting in sulin(FINS),total cholesterol(TC),serum triglyceride(TG),high density 1 ipoprotein(HDL-C),low density lipoprotein(LDL-C)and free fatty acids(FF A),calculating the indexes of homeostasis model assessment of insulin re sistant(HOMA-IR),function of pancreas islet β cell(HOMA-β)and insulin s ensitivity(ISI).To isolate pancreatic tissue under aseptic environment o n the ice rapidly.Each group take another 6 rats’ pancreatic tissue to b e fixed with 4%paraformaldehyde.Immunohistochemistry method was perform ed with GRP78,ATF4,CHOP.Western blotting was performed to detect PERK a nd P-PERK proteins expression.TUNEL were used to detect islet cell apopt osis.We used SPSS22.0 for statistical analysis of experimental data.The experimental data was expressed as x±s.One-way variance(ANOVA)was us ed for comparing means among groups.Results(1)Health state:Comparing with normal group,the model group presented obviously polydipsia,polyphagia,polyuria,emaciation symptoms and its hair was grey,dull and hairfalling,the rats dropping was hard and dry and sometimes thin stool soft,and always get perineal infection.while Modified Banxia Xiexin Decoction group and metformin group were with better presentations after treatment.(2)Effect on weight:Before treatment,the weight of T2DM rats was lower than that of normal group(P<0.05).After treatment,the weight of m odel group was lower than the nomal one(P<0.01),while the weight of Modif ied Banxia Xiexin Decoction group and metformin group increased(P<0.05).(3)Effect on FBG、FINS、HOMA-IR、HOMA-β、ISI:One week after the model,the FBG,FINS,HOMA-IR of model group was higher than the normal group(P<0.01),while the ISI and HOMA-β were lower(P<0.01).After treatment,ompareing with model group,the FBG of Modified Banxia Xiexin Decoction group and metformin group decreased,while the HOMA-β rised(P<0.05).Modified Banxia Xiexin Decoction group and metformin group could reduce HOMA-IR,but enhance ISI(P<0.05).(4)Effect on blood lipid:Comparing with normal group,the TC,TG,LDL-C and FFA of model group improved while HDL-C decreased(P<0.01).The Modified Banxia Xiexin Decoction group and metformin group could lower serum TC,TG,LDL-C,FFA and increase HDL-C in contrast with model group(P<0.05).(5)The expression of PERK and P-PERK:All the group were no statistical significance comparing the expression of PERK protein(P>0.05).The expression of P-PERK protein in pancreatic tissue of rats and the ratio of PERK to P-PERK in the model group was higher than the blank group(P<0.01)which had statistical significance.The Modified Banxia Xiexin Decoction group and metformin group were significantly decreased compared with the model group(P<0.01),but still higher than the blank group(P<0.01)which had statistical significance.(6)The expression of GRP78,ATF4,CHOP:The expression of GRP78,ATF4,CHOP protein in pancreatic tissue of rats in the model group was higher than the blank group(P<0.05)which had statistical significance.The Modified Banxia Xiexin Decoction group and metformin group were decreased significantly compared with model group(P<0.01),but was still higher than blank group(P<0.01),which had statistical significance.(7)Pancreatic cells apoptosis:Comparing with the normal group,the apoptosis rate of islet cells in each T2DM rats group was increased,and the model group was significantly increased.Compared with the model group,the apoptosis rate of islet cells in the metformin group and the modified Banxia Xiexin Decoction group decreased(P<0.01),which had statistical significance.Conclusion(1)There is ERS on pancreatic tissue of T2DM rats established by high-fat and high-sugar diet combining with low dose STZ.(2)Modified Banxia Xiexin Decoction can improve the general state of type 2 diabetic rats,regulate glucose and lipid metabolism,improve insulin resistance and improve the quality of life of diabetic rats.(3)Banxia Xiexin Decoction can inhibit endoplasmic reticulum stress signaling pathway of PERK-ATF4-CHOP,reduce the expression of GRP78,P-PERK,ATF4,CHOP protein,reduce the apoptosis of pancreatic cells,protect the islet cell function,delay the progress of type 2 diabetes. |
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