白细胞介素10基因转移抑制肾小球系膜细胞诱生炎症细胞因子  

Effects of interleukin 10 gene transfer on the secretion of inflammatory cytokines by rat glomerular mesangial cells induced by LPS in vitro

作　　者：李锦华[1] 余学清[1] 陈永雄[1] 张仕光[1] 

机构地区：[1]中山医科大学附属第一医院

出　　处：《中国病理生理杂志》1999年第4期321-324,共4页Chinese Journal of Pathophysiology

摘　　要：目的：通过建立白细胞介素１０（ＩＬ－１０）的重组逆转录病毒载体基因转移系统，观察ＩＬ－１０对脂多糖（ＬＰＳ）诱导的大鼠肾小球系膜细胞（ｇｌｏｍｅｒｕｌａｒｍｅｓｅｎｇｉａｌｃｅｌ，ＧＭＣ）中细胞因子的产生及其基因表达的影响。方法：通过构建的重组逆转录病毒载体ｐＬＸ（ＩＬ－１０）ＳＮ将外源基因ＩＬ－１０转移至大鼠ＧＭＣ：（１）应用聚合酶链反应（ＰＣＲ），反转录聚合酶链反应（ＲＴ－ＰＣＲ）和ＥＬＩＳＡ检测ＩＬ－１０基因的整合和表达；（２）以ＲＴ－ＰＣＲ观察ＩＬ－１０基因转移对ＬＰＳ诱导的ＧＭＣ肿瘤坏死因子－α（ＴＮＦ－α）的ｍＲＮＡ表达的影响，以ＥＬＩＳＡ测定白细胞介素－１β（ＩＬ－１β）和ＴＮＦ－α的蛋白质表达。结果：外源性ＩＬ－１０基因已整合到靶细胞染色体ＤＮＡ并有效地表达，它能抑制ＬＰＳ诱生ＧＭＣ过度产生ＩＬ－１β，ＴＮＦ－α。结论：外源性ＩＬ－１０基因可以转移到ＧＭＣ并稳定表达，它能抑制ＧＭＣ炎症效应中细胞因子的产生及其基因表达。AIM:To investigate the effects of interleukin 10 gene transfer on the secretion of inflammatory cytokines in rat glomerular mesangial cells (GMC) induced by LPS in vitro. METHODS:We used constructed retroviral vector pLX(IL-10) SN to transfer exogenous gene interleukin 10 into rat GMC after gene transfer 72 hours, DNA and RNA were prepared from rat transfected GMC for the polymerase chain reaction (PCR) and the reverse transcription polymerase chain reaction (RT-PCR).Expression of IL-10 in transfected GMC was checked by ELISA. RT-PCR and ELISA were performed to observe the effects of interleukin 10 gene transfer on the secretion of inflammatory cytokines in rat GMC cells induced by LPS in vitro .RESULTS:Exogenous IL-10 gene had been integrated into the chromosal DNA of the transfected GMC cells. RT-PCR and ELLSA showed that exogenous IL-10 gene was expressed in the transfected GMC after transfection 48 hours and lasting for over 28 days. Exogenous IL-10 gene transfer can inhibit the secretion of the inflammatory cytokines IL-1β and TNF-α in the transfected GMC at mRNA and protein level induced by LPS.CONCLUSION:Exogenous interleukin 10 gene transfer can inhibit the secretion of inflammatory cytokines in GMC.

关 键 词：白细胞介素10 肾小球系膜 炎症 基因转染 

分 类 号：R692.6[医药卫生—泌尿科学] R392.12[医药卫生—外科学]