石斛合剂对糖尿病大鼠糖脂代谢及其相关通路的影响

Effect of Dendrobium Mixture on Glucose and Lipid Metabolism and Related Pathways in Diabetic Rats

目的基于iTRAQ技术检测肝脏蛋白组学,探讨石斛合剂对糖尿病糖脂代谢及相关通路的影响。方法将40只Wistar大鼠随机分为正常组10只和造模组30只。正常组予普通饲料喂养,造模组予高脂高糖饲料喂养6周后,2次腹腔注射链脲佐菌素(STZ)25 mg/kg造模。造模成功后将造模组按随机数字表法分为模型组、石斛合剂组和二甲双胍组,每组10只。石斛合剂组以石斛合剂1号方灌胃7 d,继以石斛合剂2号方灌胃3 d,如此循环灌胃60 d;二甲双胍组以二甲双胍溶液灌胃,模型组、正常组用生理盐水灌胃,每组均灌胃60 d。各组大鼠于末次给药后,禁食12 h后采血检测空腹血糖(FBG)、总胆固醇(TC)和甘油三酯(TG);提取大鼠肝组织蛋白,采用iTRAQ技术联用液相串联质谱(LC-MS/MS)鉴定并分析各组的差异蛋白,并进行差异蛋白代谢通路富集分析。结果与模型组比较,石斛合剂组FBG、TC、TG显著下降(P<0.05或0.01);肝组织差异蛋白涉及了糖异生、糖酵解代谢途径及胰岛素信号通路,这些代谢途径及信号通路异常可直接或间接影响糖尿病糖脂代谢。结论石斛合剂可通过多层次、多途径影响糖尿病大鼠糖脂代谢,这为深入研究石斛合剂对糖尿病的作用机制奠定了基础。

Objective:To explore the effect of Dendrobium mixture on glucose and lipid metabolism and related pathways in diabetes,and to analyze liver proteomics based on iTRAQ technology.Methods:Forty Wistar rats were randomly divided into normal group(n=10)and modeling group(n=30).The model was established by intraperitoneal injection of streptozotocin(25 mg/kg)twice after feeding with high-fat and high-sugar diet for 6 weeks.The normal group was fed with normal diet.The rats with successful modeling were randomly divided into model group,Dendrobium mixture group and metformin group,with 10 rats in each group.The Dendrobium mixture group was given Dendrobium mixtureⅠformula by gavage for 7 days,followed by Dendrobium mixtureⅡformula for 3 days,and the circulation intervention was conducted in this way for 6 times.The metformin group was given metformin solution by gavage,while the model and normal groups were given normal saline by gavage.All groups were given by gavage for 60days.After the last administration,the rats in each group were fasted for 12 hours and then blood samples were collected to detect fasting blood glucose(FBG),total cholesterol(TC)and triglyceride(TG).iTRAQ technology combined with liquid chromatographytandem mass spectrometry(LC-MS/MS)were used to identify and analyze the differential proteins in liver tissues of each group,and the differential protein metabolic pathway was enriched and analyzed.Results:Compared with the model group,the levels of FBG,TC and TG in the Dendrobium mixture group significantly decreased(P<0.05 or P<0.01).The hepatic differential proteins identified were involved in gluconeogenesis,glycolysis metabolic pathways and insulin signaling pathways,and the abnormalities of these metabolic pathways and signaling pathways can directly or indirectly affect the glucose and lipid metabolism in diabetes.Conclusion:Dendrobium mixture can affect the metabolism of glucose and lipid in diabetes rats through multiple levels and pathways,which lays a foundation for further research on the mechanism of Dendrobium mixture on diabetes.