白细胞介素10抑制枯否细胞诱导的肝星状细胞激活的研究  

Interleukin-10 inhibits the activation of cultured rat hepatic stellate cells induced by Kupffer cells

作　　者：翁山耕[1] 冷希圣[1] 魏玉华[1] 彭吉润[1] 张佑彬[1] 吕建锋[1] 杜如昱[1] 

机构地区：[1]北京大学人民医院外科

出　　处：《中华医学杂志》2002年第2期104-107,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目 (3 9970 72 2 )

摘　　要：目的　研究白细胞介素 10 (IL 10 )对与枯否氏细胞 (KC)共培养的肝星状细胞 (HSC)激活的影响 ,并初步探讨其作用机制。方法　采用肝脏离体胶原酶灌注消化 ,及密度梯度离心的方法来分离培养HSC和KC ;将原代培养 2d的HSC和KC分为HSC单独培养、KC单独培养和HSC与KC共培养 3大组 ,分别加 2ng/ml和 2 0ng/ml的IL 10处理。 2d后 ,分别采用3 H 胸腺嘧啶脱氧核苷 (3 H TdR)掺入法检测HSC的增殖 ,Western印染法检测HSC中α 平滑肌肌动蛋白 (α SMA)的表达和酶联免疫吸附分析 (ELISA)法检测培养上清中肿瘤坏死因子 α(TNF α)的含量。结果　IL 10对单独培养的HSC的增殖 (P >0 .0 5 )和α SMA表达 (P >0 .0 5 )无明显影响。共培养组HSC的增殖和α SMA表达分别增加了 2 .4倍和 6倍 ;2ng/ml和 2 0ng/ml的IL 10分别使共培养组HSC的增殖降低了 2 3%和 33% ,α SMA表达降低了 35 %和 4 9% ,均呈浓度依赖性 ,和HSC单独培养组相比 ,差异有显著意义 (P <0 .0 5或P <0 .0 0 1)。HSC单独培养组未检测出TNF α ;共培养组的TNF α量比KC单独培养组增加了 74 %(P <0 .0 1) ;2ng/ml和 2 0ng/ml的IL 10分别使共培养组的TNF α量降低了 2 7% (P <0 0 1)和 36 % (P<0 0 1) ,使KC单独培养组的TNF α量降低了 2 9% (P <0 0 5 )和Objective To investigate the effect of interleukin 10 (IL 10) on the activation of cultured rat hepatic stellate cells (HSC) induced by Kupffer cells (KC) and relevant mechanisms. Methods HSC and KC were isolated and purified from rat liver by collagenase IV perfusion and density gradient centrifugation with Nycodenz. After primary culture for 2 days, HSC and KC were divided randomly into three groups: HSC group, KC group and HSC+KC group (coculture group), then were stimulated by IL 10 of concentrations of 2ng/ml or 20ng/ml respectively. After 2 days, the proliferation of HSC were determined with 3?H TdR incorporating test, the expression of α smooth muscle actin (α SMA) in HSC was detected by Western blotting, and tumor necrosis factor α(TNF α) protein concentration in the supernatant was determined by ELISA. Results IL 10 showed no significant effect on HSC proliferation ( P >0 05) and α SMA expression ( P >0 05) in HSC group. The levels of proliferation and α SMA expression of HSC in coculture group were 2 4 times( P <0 001) and 6 times( P <0 001) higher respectively than that in HSC groups. In coculture group, HSC proliferation and α SMA expression were decreased by 23%( P <0 001)and 35%( P <0 05)respectively after stimulation of 2ng/ml IL 10, and were decreased by 33%( P <0 001) and 49%( P <0 05)respectively after stimulation of 20ng/ml IL 10 in a dose dependent way. TNF α was not detected in HSC group. The level of TNF α in HSC+KC coculture group was 74% higher than that in KC group( P <0 01). 2 ng/ml and 20ng/ml IL 10 reduced the TNF α levle by 27%( P <0 01)and 36%( P <0 01)respectively in coculture group, and reduced the TNF α level by 29%( P <0 05)and 42%( P <0 01)respectively in KC group in a dose dependent way. Conclusion IL 10 reduces the level of TNF α secreted by KC dose dependently. Through reducing cytokine production by KC, IL 10 inhibits the activation of cultured rat HSC induced by KC, which may play a protective

关 键 词：HSC Α-SMA IL-10 共培养 白细胞介素10 肝星状细胞 增殖 浓度 诱导 消化 

分 类 号：R575.2[医药卫生—消化系统]