黄芪多糖联合尿干细胞移植对大鼠2型糖尿病的影响

Hai-Rong Zhao, Jifeng Liu, Chunmei Liu
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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. \n \n \nResults \nThe 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). \n \n \nConclusions \nAstragalus 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. 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引用次数: 0

摘要

目的研究黄芪多糖联合hucs移植对2型糖尿病大鼠的影响。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。将成功造模的100只大鼠随机分为糖尿病组、黄芪多糖处理组、黄芪多糖+黄芪多糖处理组,每组25只。治疗2周后,测定各组空腹血糖浓度、胰岛素、c肽浓度及体重变化。荧光显微镜观察pkh -26标记的胰腺内皮细胞在大鼠胰腺组织中的分布和存活情况。采用TUNEL法检测大鼠胰岛细胞凋亡情况。采用实时荧光定量PCR和Western Blot检测TGF-β/Smad信号通路相关基因在大鼠胰腺组织中的表达。结果黄芪多糖处理组、黄芪多糖处理组、黄芪多糖+黄芪多糖处理组大鼠FBG浓度均显著降低,联合处理组大鼠FBG浓度显著低于黄芪多糖组、黄芪多糖组,差异均有统计学意义(P<0.05)。与糖尿病组比较,黄芪多糖治疗组、hUSCs治疗组和联合治疗组大鼠胰岛素浓度、c肽浓度和体重均显著升高,且联合治疗组显著高于黄芪多糖治疗组和hUSCs治疗组,差异均有统计学意义(P<0.05)。荧光显微镜检查结果显示,联合治疗组各高倍视场PKH-26阳性hUSCs数量为74.64±9.75个,显著高于hUSCs治疗组的43.64±5.83个,差异有统计学意义(P<0.05)。与糖尿病组比较,黄芪多糖处理组和hUSCs处理组胰岛细胞凋亡率降低,胰腺组织TGF-β1、Smad3、Smad7 mRNA和蛋白的相对表达量也显著降低(均P<0.05)。联合治疗组降低更为显著,差异均有统计学意义(P<0.05)。结论黄芪多糖联合hUSCs移植可有效降低FBG浓度,增加胰岛素、c肽浓度和体重,减少胰岛组织凋亡,这可能与降低胰腺组织TGF-β/Smad有关。信号通路参与抑制炎症反应。关键词:黄芪多糖;人尿源性干细胞;移植;2型糖尿病;老鼠
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Effect of astragalus polysaccharide combined with urine stem cell transplantation on type 2 diabetes in rats
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. Key words: Astragalus polysaccharide; Human urine-derived stem cells; Transplantation; Diabetes mellitus, type 2; Rat
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