黄芪多糖联合人尿液干细胞移植对2型糖尿病大鼠的影响

Effect of astragalus polysaccharide combined with urine stem cell transplantation on type 2 diabetes in rats

目的研究黄芪多糖联合人尿液干细胞(hUSCs)移植对2型糖尿病大鼠的改善作用。方法随机选取25只SD大鼠作为正常对照组,其余105只SD大鼠建立2型糖尿病模型。将建模成功的100只大鼠随机分为糖尿病组、黄芪多糖治疗组、hUSCs治疗组和黄芪多糖+hUSCs联合治疗组,每组25只。治疗后2周,检测各组大鼠的空腹血糖(FBG)浓度、胰岛素和C肽质量浓度及体质量变化;荧光显微镜观察PKH-26标记hUSCs在大鼠胰腺组织中的分布和存活情况;TUNEL法检测大鼠胰岛细胞的凋亡情况;实时荧光定量PCR和蛋白质印迹法(Western Blot)检测大鼠胰腺组织中转化生长因子-β(TGF-β)/Smad信号通路相关基因的表达变化。结果与糖尿病组相比,黄芪多糖治疗组、hUSCs治疗组和黄芪多糖+hUSCs联合治疗组大鼠的FBG浓度均明显下降,且联合治疗组明显低于黄芪多糖治疗组和hUSCs治疗组,差异均具有统计学意义(均P<0.05);同时,3组大鼠的胰岛素质量浓度、C肽质量浓度及体质量明显升高,且联合治疗组明显高于黄芪多糖治疗组和hUSCs治疗组,差异均具有统计学意义(均P<0.05)。荧光显微镜观察结果显示,联合治疗组的PKH-26阳性hUSCs数[(74.64±9.75)个/视野]明显高于hUSCs治疗组[(43.64±5.83)个/视野],差异具有统计学意义(P<0.05)。与糖尿病组相比,黄芪多糖治疗组和hUSCs治疗组大鼠胰岛细胞凋亡率降低,胰腺组织中TGF-β1、Smad3、Smad7的mRNA和蛋白相对表达量也明显降低(均P<0.05),联合治疗组降低更为明显,差异均具有统计学意义(均P<0.05)。结论黄芪多糖联合hUSCs移植可有效降低2型糖尿病大鼠的FBG浓度,升高胰岛素、C肽质量浓度及体质量,减少胰岛组织细胞凋亡,这可能与降低胰腺组织中的TGF-β/Smad信号通路进而抑制炎症反应有关。

Objective To study the effect of astragalus polysaccharides combined with hUSCs transplantation on type 2 diabetic rats.Methods Twenty-five SD rats were randomly selected into the normal control group,and the remaining 105 SD rats were used to establish type 2 diabetes model.The 100 rats successfully modeled were randomly divided into the diabetes group,astragalus polysaccharide treatment group,hUSCs treatment group,and astragalus polysaccharide+hUSCs treatment group,with 25 rats in each group.After 2 weeks of treatment,the FBG concentration,insulin and C-peptide concentrations,and body weight changes were measured in each group.The distribution and survival of PKH-26-labeled hUSCs in rat pancreatic tissue were observed by fluorescence microscopy.TUNEL method was used to detect the apoptosis of rat islet cells.Real-time quantitative PCR and Western Blot were used to detect the expression of TGF-β/Smad signaling pathway-related genes in rat pancreatic tissue.Results The FBG concentration of rats in the astragalus polysaccharide treatment group,hUSCs treatment group and astragalus polysaccharide+hUSCs treatment group were significantly decreased,and that in the combination treatment group was significantly lower those in the astragalus polysaccharide group and hUSCs group,and the differences were statistically significant(all P<0.05).Compared with the diabetic group,the insulin concentration,C-peptide concentration and body weight in the astragalus polysaccharide treatment group,hUSCs treatment group and combination treatment group rats were significantly increased,and those in the combination treatment group was significantly higher than those in the astragalus polysaccharide treatment group and in the hUSCs treatment group,the differences were statistically significant(all P<0.05).The results of fluorescence microscopy showed that the number of PKH-26 positive hUSCs in the combined treatment group was 74.64±9.75 in each high power field,which was significantly higher than that in the hUSCs treatment group(43.64±5.83),the difference was statistically significant(P<0.05).Compared with the diabetic group,the apoptotic rates of islet cells in the astragalus polysaccharide treatment group and the hUSCs treatment group were reduced,and the relative expressions levels of mRNA and protein of TGF-β1,Smad3,and Smad7 in the pancreatic tissue were also significantly reduced(all P<0.05).The reduction was more significant in the combination treatment group,and the differences were statistically significant(all P<0.05).Conclusions Astragalus polysaccharide combined with hUSCs transplantation can effectively reduce the FBG concentration,increase the concentration of insulin,C-peptide and body weight,reduce the apoptosis of pancreatic islet tissue,which may be related to the reduction of TGF-β/Smad in pancreatic tissue.Signaling pathways are involved in suppressing the inflammatory response.