内毒素诱生大鼠腹腔巨噬细胞IL-12 mRNA表达调控及大黄酸对其作用研究  

Regulation of IL-12 mRNA Expression in Lipopolysaccharide and Rhei Stimulated Rat Peritoneal Macrophages

作　　者：赵琪[1,2] 崔乃强[1,2] 张立冬[1,2] 

机构地区：[1]天津市中西医结合急腹症研究所 [2]南开大学医学院分子生物学教研室

出　　处：《中国中西医结合外科杂志》1998年第1期1-4,共4页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：国家中医药管理局基金

摘　　要：应用ＲＴＰＣＲ及荧光探针标记技术观察大鼠腹腔巨噬细胞经内毒素活化后ＩＬ１２ｍＲＮＡ表达水平及细胞内信号传递系统和大黄酸对其调控作用。结果表明，内毒素能活化大鼠腹腔巨噬细胞使［Ｃａ２＋］ｉ水平及ＩＬ１２ｍＲＮＡ表达水平明显升高，并呈剂量依赖相关关系，随着ＬＰＳ剌激浓度的增加，ＩＬ１２ｍＲＮＡ表达水平及细胞内游离钙离子浓度逐渐增加。大黄酸能显著抑制ＬＰＳ活化的大鼠腹腔巨噬细胞ＩＬ１２ｍＲＮＡ表达水平及［Ｃａ２＋］ｉ的升高，亦呈剂量依赖相关关系，随着大黄酸浓度的增加ＩＬ１２ｍＲＮＡ表达水平及［Ｃａ２＋］ｉ逐渐降低。ＰＫＣ活化剂ＰＭＡ及Ａ２３１８７能显著提高ＩＬ１２ｍＲＮＡ的表达水平（与对照比较Ｐ＜０．００１，与ＬＰＳ组比较Ｐ＜０．０５）而ＰＫＣ抑制剂Ｃａｌ及ＳＰ能使经ＬＰＳ活化的大鼠腹腔巨噬细胞ＩＬ１２ｍＲＮＡ表达水平明显降低（与ＬＰＳ组比较Ｐ＜０．０１），大黄酸亦能显著抑制ＬＰＳ所至ＩＬ１２ｍＲＮＡ表达水平的升高。表明大鼠腹腔巨噬细胞经ＬＰＳ活化后其ＩＬ１２ｍＲＮＡ表达需经细胞内信号传递系统调控，大黄酸通过降低［Ｃａ２＋］ｉ使ＩＬ１２ｍＲＮＡ表达水平降低。he interleukin 12mRNA expression regulated by intracellular signalling pathway was detected by reverse transcriptionpolymerase chain reaction(RTPCR)and fourescent probe indentification.The concentration of intracellular free calcium[Ca^2+]i and the level of IL-12mRNA expression in rat peritoneal macropages were increased after activated by LPS.These increment showed a positive dose dependent relationship with the concentration of the stimulator,and the [Ca2+]i increment showed a positive interrelation with the IL12 mRNA expression(r=0.9183,P<0.05).Rheinic acid made a decreased slope of[Ca2+]i and leval of IL12 mRNA expression in LPS activated macrophages,and showed a negative dose dependent manner.The leval of IL12 mRNA and[Ca2+]i tested decreased with increasing doses of rheinic acid.The macrophages expressed IL12 mRNA in response to protein kinase C(PKC)activation.The level of IL12 mRNA increased significantly(P<0.001)after 4 hours incubated with phorbol myristate acetate(PMA 100nM) and A23187(50nM),decreased in calphostin C(200nM)and staurosporine(50nM)treated rat peritoneal macrophages(P<0.01).The inhibitory ability of rheinic acid(100ug/ml,P<0.01) was almost the same as the calphostin C and staurosporine on IL-12 mRNA expression in macrophages.In summary,The LPS induced IL-12 mRNA experession in rat peritoneal macrophages is regulated by intracellular signalling pathway especially PKC and the third messenger free calcium.The rheinic acid blocked the IL-12 mRNA transcription through lowering down the[Ca^2+]i and inhibition of PKC activity.

关 键 词：巨噬细胞 内毒素 MRNA 大黄酸 白细胞介素12 

分 类 号：R631.3[医药卫生—外科学] R285.5[医药卫生—临床医学]