论文部分内容阅读
目的 探讨炎症诱导启动子指导人白细胞介素 10 (hIL 10 )基因的表达及其对炎症介质产生的效应。方法 诱导性真核表达载体pSAA3hIL 10脂质体转染体外培养巨噬细胞 ,LPS(10mg/L)活化后观察巨噬细胞hIL 10的表达 (RT PCR ,ELISA法 ) ,测定肿瘤坏死因子 α(TNF α)、IL 6的产生(ELISA法 )。结果 RT PCR证明hIL 10可在巨噬细胞表达 ;巨噬细胞LPS(10mg/L)活化 12h有hIL 10的表达(2 .67μg/L) ,活化 2 4h及 4 8h的巨噬细胞上清hIL 10显著增加(1.87～5 .71倍 ) ;转基因预处理显著下调活化细胞TNF α(2 5 .0 %～ 61.4 % ,P <0 .0 5 )的产生 ,显著减少IL 6产生 (5 4 .8%～63 .9% ,P <0 .0 5 )。结论 LPS可活化 pSAA3hIL 10表达体系在巨噬细胞诱导表达 ,白细胞介素 10基因转染显著下调活化MΦ炎症介质的产生。
Objective To investigate the expression of human interleukin - 10 (hIL 10) gene induced by inflammation and its effect on inflammatory mediators. Methods The recombinant plasmid pSAA3hIL 10 was transfected into macrophages in vitro. LPS (10 mg / L) was used to observe the expression of hIL 10 in macrophages (RT - PCR, ELISA). The levels of tumor necrosis factor α TNF α), IL 6 production (ELISA method). Results The hIL 10 expression in macrophages was confirmed by RT-PCR. The expression of hIL 10 (2.67 μg / L) was induced by macrophages LPS (10 mg / L) for 12 h and activated by hIL 10 significantly increased (1.87-5.71 fold). Transgene pretreatment significantly reduced the production of TNFα (25.0% -61.4%, P <0.05) in activated cells, and significantly reduced the production of IL-6. 8% ~ 63 .9%, P <0. 05). Conclusion LPS can activate pSAA3hIL 10 expression system in macrophage induced expression of interleukin 10 gene transfection significantly down-regulated activation of MΦ inflammatory mediators.