【摘要】 目的研究白介素-13(IL-13)、前列腺素E2(PGE2)和前列环素(PGI2)在肝星形细胞(HSCs)参与肝脏纤维化中的作用。方法通过酶联免疫吸附试验(ELISA)、逆转录聚合酶链反应(RT-PCR)、蛋白质印迹法(Western blotting)、蛋白激酶C(PKC)活性检测和Brd U检测,检测了IL-13、PGE2和PGI2对转化生长因子-β1(TGF-β1)和血小板衍生生长因子(PDGF)在人HSCs中m RNA和蛋白水平的影响,以及对细胞活性及PKC活性的影响,另外,通过加入PKC的激动剂和抑制剂来研究其对TGF-β1和PDGF表达的影响。结果 IL-13、PGE2和PGI2显著提升了TGF-β1和PDGF在人HSCs中的表达量及分泌量,同时也提高了HSCs的增殖能力和细胞活性。在IL-13、PGE2和PGI2的任何一个治疗组中,PKC的活性也得到明显增高。通过ELISA、Western blotting和RT-PCR检测,发现PKC的激动剂12-豆蔻酸-13-乙酸佛波醇(PMA)明显提高了HSCs中TGF-β1和PDGF的表达量和分泌量,而PKC抑制剂钙磷酸蛋白(Calphostin C)则与此相反。同时,PMA促进HSCs的增殖活性,而Calphostin C则对HSCs的增殖产生抑制。结论 IL-13、PGE2和PGI2可能通过活化PKC来刺激人HSCs的增殖及合成并释放TGF-β1和PDGF,促进肝脏纤维化的进展。更多还原

【Abstract】 Objective To investigate the role of IL-13,PGE2 and PGI2 for participating in hepatic stellate cells(HSCs)involving hepatice fibrosis. Methods ELISA,RT-PCR,Western blot,PKC activity detection and Brd U detection weere used to detect the influence of IL-13,PGE2 and PG12 on expression levels and secretion levels of TGF-β1and PDGF in HSCs,and influence on cellular activity and PKC acivity,in addition,its influence on the TGF-β1and PDGF expression aby adding the PKC agonist and inhibitor. Results IL-13,PGE2 and PGI2 significantly increased expression levels and secretion levels of TGF-β1 and PDGF in HSCs,meanwhile increased the proliferation ability and cellular activity of HSCs. In any treatment group of IL-13,PGE2 and PG12,the PKC activity was singificantly increased. The ELISA,Western blot and RT-PCR detections found that PKC agonist PMA singificantly increased the expression levels and secretion levels of TGF-β1 and PDGF in HScs,while the PKC inhibitor calphostin C was in contrstat. Meanwhile PMA promoted the HSCs proliferation activity,while calphostin C produced the inhibition on HSCs roliferation. Conclusion IL-13,PGE2 and PGI2 stimulate HSCs proliferation,synthesization and releasing TGF-β1and PDGF possibly by activating PKC for promoting hepatic fibrosis progression.更多还原