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目的:研究雷公藤在大鼠1型糖尿病模型中对CD4+CD25+调节性T细胞(Tr)Foxp3基因表达的影响及意义。方法:将48只大鼠随机分为3组,每组各16只,Ⅱ、Ⅲ组采用小剂量多次腹腔注射链脲佐菌素(STZ)的方法制备大鼠1型糖尿病模型,Ⅰ组为正常对照。Ⅲ组在成模后给与雷公藤多甙(GTT)灌胃,1次/d,连续3个月。淋巴细胞转化试验(MTT法)检测脾淋巴细胞增殖活性;荧光定量PCR检测外周血及脾淋巴细胞Foxp3 mRNA的表达;免疫组化检测淋巴结和脾组织FOXP3蛋白的表达。结果:Ⅱ、Ⅲ组血糖水平高于Ⅰ组(P<0.01);胰岛中Ⅲ组淋巴细胞浸润程度较Ⅱ组减轻;Ⅱ、Ⅲ组淋巴细胞增殖活性均较Ⅰ组升高(P<0.01),用药后Ⅲ组较Ⅱ组降低;Foxp3 mRNA、FOXP3蛋白表达水平Ⅱ、Ⅲ组均高于Ⅰ组(P<0.05),Ⅲ组表达水平较Ⅱ组有所升高,但差别无统计学意义(P>0.05)。结论:Foxp3+Tr细胞参与了STZ诱导的大鼠1型糖尿病的发生发展;上调Tr细胞Foxp3基因的表达可能为雷公藤治疗1型糖尿病的机制之一。
Objective: To study the effect of triptolide on Foxp3 gene expression of CD4 + CD25 + regulatory T cells (Tr) in rat type 1 diabetes mellitus model. Methods: Forty-eight rats were randomly divided into three groups (16 rats in each group). Rats in group Ⅱ and group Ⅲ were given low-dose multiple intraperitoneal injection of streptozotocin (STZ) For the normal control. Group Ⅲ was given intragastric administration of tripterygium glycosides (GTT) once a day for 3 months. The proliferation of splenic lymphocytes was detected by MTT assay. The expression of Foxp3 mRNA in peripheral blood and splenic lymphocytes was detected by real-time PCR. The expression of FOXP3 in lymph nodes and spleen tissues was detected by immunohistochemistry. Results: The blood glucose level in group Ⅱ and Ⅲ was higher than that in group Ⅰ (P <0.01). The level of lymphocyte infiltration in group Ⅲ was less than that in group Ⅱ. The proliferation activity of lymphocytes in group Ⅱ and Ⅲ was higher than that of group Ⅰ (P <0.01) (P <0.05). The expression of Foxp3 mRNA and FOXP3 protein in group Ⅱ and Ⅲ were higher than that in group Ⅰ (P <0.05), while the expression in group Ⅲ was higher than that in group Ⅱ, but the difference was not statistically significant (P> 0.05). CONCLUSION: Foxp3 + Tr cells are involved in the development of type 1 diabetes mellitus induced by STZ in rats. Up-regulation of Foxp3 expression in Tr cells may be one of the mechanisms of Triptolide-treated type 1 diabetes mellitus.