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本文探讨盐酸小檗碱(BBR)对活化的MOG特异性T淋巴细胞体外增殖及其作用机制。通过建立MOG抗原特异性T淋巴细胞增殖体系,在培养初始加入梯度剂量的BBR药物,用3H摄入试验检测BBR对其增殖的影响,并采用AnnexinV染色法检测MOG特异性T细胞凋亡率,用流式细胞术检测细胞凋亡分子Fas、FasL和Caspase3的表达水平。并且采用PCR技术检测细胞抗凋亡基因Bcl-2和周期蛋白相关基因的表达水平。结果显示,在MOG特异性T淋巴细胞增殖体系中,BBR在较低浓度(1μmol/L)时就能抑制MOG特异性T淋巴细胞增殖反应(P<0.05),抑制作用呈浓度依赖性;MOG特异性T淋巴细胞的凋亡率随着药物浓度增加显著上升,细胞凋亡分子Fas、FasL和Caspase3的表达也随着药物浓度的增加而提高,PCR结果显示,抗凋亡基因Bcl-2表达下调,细胞周期G1期相关基因cyclin D1、cyclin E1、CDK2、CDK4、CDK6表达下调,G1期负调控基因p27表达上调。研究提示,BBR在体外能有效抑制MOG特异性T淋巴细胞增殖,促进其凋亡,是治疗自身免疫病的潜在药物。
This article explored the in vitro proliferation of activated MOG-specific T lymphocytes by berberine hydrochloride (BBR) and its mechanism of action. By establishing an MOG antigen-specific T lymphocyte proliferation system, a gradient dose of BBR was initially added to the culture, and the effect of BBR on its proliferation was examined by 3H uptake assay, and the apoptosis rate of MOG specific T cells was detected by Annexin V staining. Flow cytometry was used to detect the expression levels of apoptosis molecules Fas, FasL and Caspase3. The expression of anti-apoptosis gene Bcl-2 and cyclin-related genes was detected by PCR. The results showed that in the proliferation system of MOG-specific T lymphocytes, BBR could inhibit the proliferation of MOG-specific T lymphocytes at a low concentration (1 μmol/L) (P<0.05), and the inhibition was concentration-dependent; MOG The rate of apoptosis of specific T lymphocytes increased significantly with the increase of drug concentration. The expression of apoptosis molecules Fas, FasL and Caspase3 also increased with the increase of drug concentration. PCR results showed that the expression of anti-apoptosis gene Bcl-2. Down-regulation, G1 phase-related genes cyclin D1, cyclin E1, CDK2, CDK4, CDK6 downregulated, G1 phase negative control gene p27 expression upregulated. Studies have shown that BBR can effectively inhibit the proliferation of MOG-specific T lymphocytes and promote apoptosis in vitro, and is a potential drug for the treatment of autoimmune diseases.