哮喘大鼠外周血T淋巴细胞T_H1/T_H2细胞因子和PKCα的表达及银杏叶制剂对其影响  

Production of interleukin-2,4,5 and protein kinase Cα expression in peripheral blood T lymphocytes culture in vitro of asthmatic rats and the effect of Ginkgo biloba extract on them

作　　者：唐以军[1] 徐永健[1] 张珍祥[1] 倪望[1] 陈仕新[1] 高宝安[1] 叶涛[1] 曹勇[1] 杜春玲[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸内科,武汉430030

出　　处：《中华微生物学和免疫学杂志》2006年第1期67-73,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(30070332)

摘　　要：目的探讨银杏叶制剂对在哮喘免疫学发病机制中起关键作用的T淋巴细胞部分生物学功能的影响。方法14只SD大鼠随机分成哮喘和正常两组,每组7只。从每只大鼠外周血分离出T淋巴细胞进行分组培养,72h后用RT-PCR检测各组IL-2、IL-4、IL-5mRNA的表达量,Westernblot检测各组细胞膜与细胞浆蛋白激酶Cα(protein kinase Cα,PKCα)的表达比率。结果IL-2、IL-4、IL-5mRNA表达量在哮喘组分别为0.58±0.086、1.03±0.12、0.48±0.08,正常组分别为0.45±0.03、0.35±0.08、0.15±0.05,各因子两组间比较差异有统计学意义(P<0.01);哮喘组T淋巴细胞给予BN-52021干预后IL-2、IL-4、IL-5mRNA的表达量明显下降(分别为0.49±0.05、0.55±0.09、0.27±0.05,P<0.05);正常组IL-2/IL-4mRNA的比值为1.27±0.20,哮喘组为0.60±0.18,两组比较差异有统计学意义(P<0.01),其中哮喘组给予BN-52021干预后比值有所增大0.92±0.15,与未干预组比较有明显变化(P<0.01)。哮喘PMA+BN-52021干预组IL-2、IL-4、IL-5mRNA表达量分别为1.52±0.25、1.99±0.36、0.68±0.21,明显低于哮喘PMA干预组(P<0.05),而PMA+Ro31-8220组与PMA+Ro31-8220+BN-52021干预组比较差异无统计学意义(P>0.05)。哮喘空白组PKCα在T淋巴细胞胞膜与胞浆的表达量比率为0.629±0.093,显著高于正常对照组(0.056±0.012,P<0.01),BN-52021干预后哮喘组比率较未干预组明显下降0.395±0.098(P<0.05),PMA+BN-52021干预组PKCα比率为0.719±0.163,较PMA干预组(1.28±0.28)低(P<0.05);哮喘各组T淋巴细胞胞膜与胞浆PKCα表达量的比率与IL-4mRNA表达量的相关性分析(n=42,r=0.845,P<0.01)显示呈明显正相关。结论银杏叶制剂对体外培养的哮喘大鼠T淋巴细胞因子IL-2、IL-4、IL-5的分泌均具有抑制作用,而对IL-2分泌的抑制作用相对较弱;银杏叶制剂对哮喘大鼠T淋巴细胞胞膜与胞浆PKCα的表达量比率具有明显的下调作用。Objective To investigate therapeutic effect of Ginkgo biloba extract on bronchial asthma. Methods Fourteen SD rats were randomly divided into two groups, 7 rats were sensitized as asthmatic model and the others as healthy controls. Results The quantity of IL-2, IL-4 and IL-5 mRNA in blank group of asthmatic models（0.58 ± 0. 086, 1.03 ± 0.12 and 0.48 ± 0.08 respectively）, significantly increased （ P 〈 0.05）. The quantity of IL-2, IL-4 and IL-5 mRNA decreased significantly when the T lymphocytes of asthmatic model rats were cultured with BN-52021 （0.49 ± 0.05, 0.55 ± 0.09 and 0.27 ± 0.05 respectively）, P 〈 0.05. The ratio of IL-2/ IL-4 mRNA in normal rats was 1.27 ± 0.20, which was significantly higher than that in asthmatic model rats（0.60 ±0.18, P〈0.01）. BN-52021 significantly increased the ratio when the T lymphocytes of asthmatic model rats were cultured with it （ P 〈 0.01 ） . BN-5202 1 signiflcantlycounteractedthelevd of IL- 2, IL-4 and IL-5 mRNA that increased by PMA（ P 〈 0.05）, but the decreasing effect was less than the Ro31-8220, a specific inhibitor of PKCa（P 〈0.05）. BN-52021 failed to further counteract the level of IL-2, IL-4 and IL-5 mRNA increased by PMA which had significantly decreased by the Ro31-8220. The expression ratio of PKCα in membrane and cytoplasm of T lymphocytes of asthmatic model rats was significantly higher than that in normal rats（ P 〈 0.01 ）, and BN-52021 significantly decreased the ratio when the T lymphocytes were cultured with it（ P 〈0.05）. Furthermore, it also significantly counteracted the expression ratio of PKCa in membrane and eytoplasm which increased by PMA（ P 〈0.05）. The expression ratio of PKCα in membrane and cytoplasm was positively correlated with the level of IL-4 mRNA in T lymphocytes cultured in vitro in asthmatic model rats group（n=42, r=0.845, P〈0.01）. Conclusion Ginkgo biloba extract depress the expression of IL-2, IL-4 and IL-5 mRNA of asthmatic model rat T lymphocytes in vitro. However,

关 键 词：银杏叶制剂 支气管哮喘 T淋巴细胞 白细胞介素 蛋白激酶C 

分 类 号：R562.25[医药卫生—呼吸系统]