青藤碱抑制人外周血CD4^＋T淋巴细胞内NF-AT和IFN-γ表达的体外研究  

Effects of alkaloid sinomenine on expression levels of nuclear factor of activated T cells and interferon-gamma in CD4^＋ T lymphocytes of human periphery blood in vitro

作　　者：李建军[1] 罗志刚[1] 秦国庆[1] 王毅[1] 钱坤[1] 周浩[1] 

机构地区：[1]南华大学附属第二医院22病区肾移植科,湖南省衡阳市421001

出　　处：《中华器官移植杂志》2010年第8期496-499,共4页Chinese Journal of Organ Transplantation

摘　　要：目的 通过研究青藤碱（SIN）对人外周血CD4^＋T淋巴细胞内核因子-AT（NF-AT）和γ干扰素（IFN-γ）表达的影响来探讨其免疫作用机制.方法 免疫磁珠法分选人外周血CD4^＋T淋巴细胞,并分为5组进行培养.（1）阴性对照组：细胞培养液中不加入任何药物;（2）阳性对照组：细胞培养液中加入终浓度为50ng/ml的环孢素A（CsA）溶液;（3）低浓度青藤碱（L-SIN）组：细胞培养液中加入终浓度为10μmol/L的青藤碱溶液;（4）中浓度SIN（M-SIN）组：细胞培养液中加入终浓度为200 μmol/L的青藤碱溶液;（5）高浓度SIN（H-SIN）组：细胞培养液中加入终浓度为1000 μmol/L的青藤碱溶液.观察各组CD4^＋T淋巴细胞的增殖情况,蛋白印迹法检测各组细胞内NF-AT的蛋白表达,流式细胞术检测各组细胞内IFN-γ的表达水平.结果 与阴性对照组比较,M-SIN和H-SIN组明显抑制了CD4^＋T淋巴细胞的增殖（P＜0.01）.青藤碱呈浓度依赖性抑制CD4^＋T细胞内NF-AT和IFN-γ的表达（P＜0.01）,而加入CsA培养的阳性对照组NF-AT和IFN-γ表达最低.CD4^＋T淋巴细胞内NF-AT的表达与细胞增殖抑制率之间存在负相关关系（rs=-0.969,P=0.000）.结论 青藤碱抑制人外周血CD4^＋T淋巴细胞内NF-AT和IFN-γ表达;其免疫作用机制可能是通过降低NF-AT的表达以抑制T淋巴细胞的活化和增殖.Objective To study the immunosuppressive mechanism of alkaloid sinomenine （SIN） by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells （NF-AT） and interferon-gamma （IFN-γ） in CD4^＋ T lymphocytes of human periphery blood. Methods CD4^＋ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. （1） Negative control group： no medicine was added to cell culture medium; （2） Positive control group： CsA solution （final concentration： 50ng/ml） was added to cell culture media; （3） Low-concentration SIN group （L-SIN）： low-concentration SIN solution （final concentration： 10 μmol/L） was added to cell culture media; （4） Middle-concentration SIN group （M-SIN）： middle-concentration SIN solution （final concentration： 200 μmol/L） was added to cell culture media; （5） High-concentration SIN group （H-SIN）： high-concentration SIN solution （final concentration： 1000 tmol/L） was added to cell culture media. The proliferations of CD4^＋ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups （P〈0. 01 ）. SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4^＋ T lymphocytes of human periphery blood （P〈0.01）. The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4^＋ T lymphocytes of human periphery blood （rs = - 0. 969, P = 0. 000）. Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4^＋ T lymphocytes of human periphery blood probably by decreasing protein leve

关 键 词：青藤碱 免疫抑制 CD4^+T淋巴细胞内核因子-AT γ干扰素 

分 类 号：R285.5[医药卫生—中药学]