摘要
目的:研究来氟米特活性代谢物丙二酸次氮酰胺(A771726)对刀豆蛋白(ConA)刺激的胶原性关节炎(CIA)模型大鼠CD4+CD25+调节性T淋巴细胞(CD4+CD25+Tregs)体外分化的影响及其治疗类风湿性关节炎的作用机制。方法:取大鼠分为正常对照组和CIA模型组,建模后制备脾淋巴细胞悬液,再分为正常组、模型组和A771726低、中、高剂量(0.1、1、10μmol·L-1)组,各组加入ConA刺激和相应药物培养液培养48h后,以MTT法检测各组脾淋巴细胞增殖情况,流式细胞术检测各组CD4+CD25+Tregs占CD4+T淋巴细胞的比例;反转录-聚合酶链式反应检测各组CD4+CD25+Tregs特异性标志物Foxp3和转化生长因子β(TGF-β)mRNA表达情况;酶联免疫吸附测定法检测各组TGF-β浓度。结果:与正常组比较,模型组脾淋巴细胞增殖明显升高(P<0.01),CD4+CD25+Tregs占CD4+T淋巴细胞的比例、Foxp3和TGF-βmRNA表达、TGF-β浓度均明显降低(P<0.05);与模型组比较,A7717263个剂量组脾淋巴细胞增殖明显降低(P<0.05或P<0.01),A771726高剂量组CD4+CD25+Tregs占CD4+T淋巴细胞的比例明显升高,A771726中、高剂量组Foxp3和TGF-βmRNA表达、TGF-β浓度均明显升高(P<0.05或P<0.01)。结论:A771726可抑制CIA模型大鼠脾淋巴细胞增殖,诱导CD4+CD25+Tregs的分化,其机制可能与上调TGF-β的表达和分泌有关。
OBJECTIVE： To study the effect and mechanism of active metabolite malonic acid amide nitrogen （A771726） on the differentiation of CD4＋CD25＋ Tregs from collagen-induced arthritis （CIA） rats in vitro. METHODS： Rats were divided into normal control group and CIA model group. The splenocyte suspension was prepared after modeling then divided into normal group, model group and A771726 low-dose, medium-dose and high-dose groups （0.1,1,10 μmol-L-1）. Those groups were cultured in culture solution of ConA and relevant medicine for 48 h. The proliferation of spleen lymphocytes from CIA rats was checked with MTT assay; immunofluorescence flow cytometry was used to detect the proportion of CD4＋CD25＋Tregs in CD4＋T; RT-PCR was used to detect the expression of Foxp3 mRNA and TGF-β mRNA; ELISA was used to detect the content of TGF-β. RESULTS： Compared with normal group, the proliferation of ConA-induced splenocytes of CIA rats in model group increased significantly （P〈0.01）, while the proportion of CD4＋CD25＋ Tregs, the expression of Foxp3 and TGF-β mRNA and the content of TGF-β decreased significantly （P〈0.05）. Compared with model group, the proliferation of ConA-induced splenocytes in A771726 groups decreased significantly （P〈0.05 or P〈0.01）, while the proportion of CD4＋CD25＋ Tregs in CD4＋T ofA771726 high-dose group increased significantly, and the expression of Foxp3 and TGF-β mRNA and the content of TGF-β in A771726 medium-dose and high-dose groups increased significantly （P〈0.05 or P〈0.01）. CONCLUSION： A771726 could inhibit the proliferation of splenocytes of CIA rats, and facilitate the differentiation of CD4＋CD25＋Tregs, which may be related to the up-regulation of the expression and secretion of TGF-β.
出处
《中国药房》
CAS
CSCD
2012年第1期34-36,共3页
China Pharmacy
