健脾消渴方改善2型糖尿病大鼠药效学研究

Pharmacodynamic Study on Jianpi Xiaoke Decoction(健脾消渴方)in Improving Type 2 Diabetic Rats

目的评价健脾消渴方对2型糖尿病大鼠的药效,并探讨其可能的作用机制。方法40只SD大鼠随机分为正常对照组、模型对照组、健脾消渴方组及盐酸二甲双胍组,每组10只,STZ+高脂饮食建立2型糖尿病大鼠模型,给予相应药物,持续4周。实验结束后取材,考察大鼠空腹血糖变化情况;ELISA法检测大鼠血清中白细胞介素-1(interleukin-1β,IL-1β)、白细胞介素-10(interleukin-10,IL-10)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、D-氨基酸氧化酶(D-amino acid oxidase,DAO)、D-乳酸含量;考察血清血脂四项[甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)]含量;HE染色法观察各组大鼠胰腺及回肠的组织形态学改变;免疫荧光法检测大鼠回肠L细胞的免疫荧光法检测大鼠回肠L细胞的胰高糖素样肽-1(glucagon-like peptide-1,GLP-1)阳性表达;Western blotting法检测大鼠结肠组织中紧密连接蛋白(occludin)、闭锁小带蛋白-1(zonula occludens-1,ZO-1)、黏蛋白2(Muc2)蛋白相对表达。结果健脾消渴方可有效降低2型糖尿病大鼠的空腹血糖,下调血清中IL-1β、TNF-α含量,上调IL-10含量,抑制DAO、D-乳酸含量;同时,显著性下调TG、TC、LDL-C含量,但在提高HDL-C含量方面上仅具有趋势性;此外,健脾消渴方可改善大鼠胰腺及回肠损伤,提高GLP-1阳性表达,并有效降低2型糖尿病大鼠结肠occludin、ZO-1、Muc2蛋白相对表达。结论健脾消渴方对2型糖尿病大鼠具有较好的治疗效果,其作用可能与改善血清炎症、调节血脂,减轻肠道功能损伤,改善肠道屏障通透性与保护肠道屏障生理结构等作用有关。

Objective To evaluate the effect of Jianpi Xiaoke Decoction(健脾消渴方)on type 2 diabetic rats and explore its possible mechanism.Methods 40 cases SD rats were randomly divided into normal control group,model control group,Jianpi Xiaoke Decoction group and positive medicine group,10 rats in each group.STZ+high fat diet was used to establish type 2 diabetic rat model,and corresponding medicine was given for 4 weeks.After the end of the experiment,the rats were sampled to investigate the changes of fasting blood glucose;the expression levels of IL-1β,IL-10,TNF-α,DAO and D-lactic acid in serum and the changes of serum lipids(TG,TC,HDL-C and LDL-C)were detected by ELISA;the histomorphological changes of pancreas and ileum were observed by HE staining;the GLP-1 positive expression in L cells of ileum was detected by immunofluorescence method,and the relative expression of occludin,ZO-1 and Muc2 proteins in colon tissue of rats was detected by western blotting.Results Jianpi Xiaoke Decoction could effectively reduce fasting blood glucose,down-regulate IL-1β,TNF-αinflammatory factors expression in serum,promote IL-10 content expression,inhibit DAO,D-lactic acid content expression,significantly down-regulate TG,TC,LDL-C expression,but only had a tendency to increase HDL-C expression;meanwhile,Jianpi Xiaoke Decoction could improve pancreatic and ileal injury,increase GLP-1 positive expression,and effectively reduce the relative expression of occludin,ZO-1,Muc2 protein in colon of type 2 diabetic rats.Conclusion Jianpi Xiaoke Decoction has a better improving effect on type 2 diabetic rats,which may be related to improving serum inflammation,regulating the expression of lipid content,alleviating intestinal function damage,improving intestinal barrier permeability and protecting intestinal barrier physiological structure.