摘要
目的通过体外实验探讨钙调神经磷酸酶抑制剂环孢菌素-A对男性结核性胸膜炎免疫调节的影响。方法选取30例确诊结核性胸膜炎的男性患者为受试对象,于治疗前经无菌术采集胸腔积液,以人淋巴细胞分离液及贴壁培养法分离其中细胞;细胞提纯后经免疫细胞化学染色法检验细胞类型,调细胞浓度为1×106/ml。将每例患者采集的胸腔积液样本等分为3份,分别设立为空白对照组(未予以药物干预)、多抗甲素组(孔内依次加入浓度为0.1μg/ml、10.0μg/ml、50.0μg/ml的多抗甲素)和实验组(孔内依次加入浓度为10.0μg/ml、30.0μg/ml、50.0μg/ml的环孢菌素-A);十二孔培养板内培养液均为2ml,三组均分别设立3个复孔对应3个不同时间点(0h、24h和72h)。经酶联免疫吸附法(ELISA)检测不同时间点各组上清液及培养液中γ-干扰素(IFN-γ)、转化生长因子-β(TGF-β)、白细胞介素-4(IL-4)、白细胞介素-12(IL-12)的浓度差异。结果免疫化学ABC法鉴定结果显示,30例受试者胸腔积液中分离所得的细胞多数着色,且表现为棕色,有CD4+抗原决定簇,确定为CD4+T淋巴细胞。24h和72h时,三组样本IFN-γ、IL-12水平均较0h显著降低(P<0.05),但72h检测结果高于24h时,差异有统计学意义(P<0.05);实验组和多抗甲素组24h和72h时,不同浓度药物处理后IFN-γ、IL-12水平均明显高于空白对照组,差异有统计学意义(P<0.05);24h和72h时,三组样本TGF-β、IL-4水平均较0h显著降低(P<0.05),且72h检测结果明显低于24h时,差异有统计学意义(P<0.05);实验组和多抗甲素组24h和72h时,不同浓度药物处理后TGF-β、IL-4水平均明显低于空白对照组,差异有统计学意义(P<0.05)。结论环孢菌素-A对男性结核性胸膜炎患者有一定免疫调节作用,有利于病情转归,可在临床进一步推广。
Objective To explore the effect of calcineurin inhibitor cyclosporin -A on immune regulation of patients with tuberculous pleurisy through in-vitro. Methods 30 male patients with confirmed tuberculous pleurisy were selected as subjects. The pleural effusion was collected by aseptic technique before treatment. The cells were isolated by human lymphocyte separating medium and adherent culture method. After cell purification, the cell types were tested by immunocytochemistry staining, and the cell concentration was 1 × 10^6/mi. The pleural effusion samples were equally divided into blank control group （without drug intervention）, polyresistin control group （concentrations were 0.1μg/ml, 10.0μg/ml and 50.0μg/ml） and the experimental group （concentrations were 10.0μg/ml, 30.0μg/ml and 50.0μg/ml）. The culture medium in twelve-hole culture plate was 2ml. In blank control group, different concentrations of polyresistin control group and experimental group, 3 accessory foraminas were set up, corresponding to 3 different time points （0h, 24h and 72h）. By enzyme-linked immunosorbent assay （ELISA）, the concentrations of interferon gamma （IFN-γ）, transforming growth factor beta （TGF-γ）, interleukin 4 （IL-4） and interleukin-12 （IL-12） in supernatant and culture medium were detected at different time points. Results The results of immunochemical ABC showed that the majority of cells in the pleural effusion of 30 patients were brown, with CD4＋ antigenic determinant, and were identified as CD4＋T lymphocytes. At 24h and 72h, the levels of IL-12 and IFN-γ in the three groups were significantly lower than those at Oh （P〈0.05） but the test results at 72h were significantly higher than those at 24h （P〈0.05）. At 24h and 72h, after treated with different concentrations of drugs, the levels of IL-12 and IFN-γ in the experimental group and the polyresistin control group were significantly higher than those in the blank control group （P〈0.05）. At 24h and 72h, the levels of TGF-β and IL-4 in the three groups were significantly lower than those at Oh （P〈0.05） and the test results at 72h were significantly lower than those at 24h （P〈0.05）; At 24h and 72h, after being treated with different concentrations of drugs, the levels of TGF-β and IL-4 in the experimental group and the polyresistin control group were significantly higher than those in the blank control group （P〈0.05）. Conclusion Cyclosporine-A has some immunomodulatory effects on patients with tuberculous pleurisy, and the prognosis is favorable.
出处
《西部医学》
2017年第1期27-31,共5页
Medical Journal of West China
基金
四川省卫生厅科研课题(120327)
