丹参对肝星状细胞丝裂原活化蛋白激酶通路的抑制作用  

Effects of radix salviae miltiorrhizae (RSM) on MAPK pathway in hepatic stellate cells (HSCs)

作　　者：李校天[1] 杨书良[2] 王军民[3] 柳继兴[4] 赵如同[2] 

机构地区：[1]河北工程学院临床医学院消化内科,邯郸056029 [2]河北工程学院临床医学院病理科,邯郸056029 [3]河北省人民医院消化科 [4]河北工程学院临床医学院检验科,邯郸056029

出　　处：《解放军医学杂志》2006年第1期22-24,共3页Medical Journal of Chinese People's Liberation Army

基　　金：河北省科技厅基金资助项目(02276103D8)

摘　　要：目的探讨肝星状细胞丝裂原活化蛋白激酶(HSCsMAPK)通路中ERK、JNK的活化情况,以及丹参药物血清对磷酸化细胞外信号调节蛋白激酶(PERK)、磷酸化氨基末端蛋白激酶(PJNK)表达的影响。方法用大鼠制备肝纤维化模型。造模后,药物组以每天20ml/kg剂量分2次灌服丹参;对照组灌以等量生理盐水。连续给药6天后经下腔静脉取血并分离血清。采用盲法,用上述10%药物血清培养HSCs24h后,用Westernblot方法检测各组HSCsPERK、PJNK蛋白表达水平。结果各组在约44kD、42kD的位置上均出现杂交带,表明活化的HSCs同时表达PERK1和PERK2;在约46kD、54kD的位置上均出现杂交带,表明HSCs同时表达PJNK1和PJNK2。PERK蛋白表达水平,肝纤维化大鼠经丹参药物血清干预后较对照组显著降低(18.03%±3.99%vs28.16%±5.37%,P<0.01),正常大鼠经丹参药物血清干预后较正常大鼠对照组也显著减少(18.17%±4.02%vs26.35%±8.22%,P<0.05)。PJNK蛋白表达水平,肝纤维化大鼠经丹参药物血清干预后较对照组显著降低(17.70%±2.83%vs32.66%±7.08%,P<0.01),正常大鼠经丹参药物血清干预后较正常大鼠对照组也显著减少(19.53%±2.48%vs29.20%±6.27%,P<0.05)。结论活化的HSCs中ERK、JNK保持着较高的磷酸化水平。提示两者介导的信号转导通路是HSCs活化和增殖的重要途径之一;阻断MAPK信号通路可能是丹参治疗肝纤维化的重要作用途径之一。Objective To observe two main subtypes of MAPK family-ERK and JNK activation in HSCs and investigate the effects of the serum of Radix Salviae Militiorrhizae treated animals on the activation of ERK and JNK in HSCs. Methods HSCs were cultured in vitro. After the model of hepatic fibrosis was replicated in SD rats, Radix Salviae Militiorrhizae decoction was given via gastrogavage to the rats of treatment groups to obtain serum containing the drug. The dose was 10 times of dose per kg per day for adults in 2 divided doses for 6 consecutive days, while the same volume of 0. 9% NaCl was given to the rats of non-treatment groups, on the 7th day, the routine dose was orally given again; the blood sample was drawn from the vena cava after 2 hour; the serum was isolated and inactivated with water bath at 56℃ ; finally, the serum was filtered to eliminate bacteria. Just before using the serum, RPMI-1640 culture medium was added to prepare culture media of 10% drug-containing serum, which was incubated with the subcultured H~：s. The experiment was divided into the following groups. A： serum of extracted from normal rats; B： medicated serum of RSM extracted from normal rats; C： serum from CCl4 induced liver fibrosis rats; D： medicated serum of RSM extracted from CCl4-induced liver fibrosis rats. After 24h incubation with a hove every group serum which were added blindly to HSCs, P-ERK and P-JNK were detected by Western blot respectively. Results Both RSM Pharmacological serums decreased P-ERK and P-JNK in HSCs significantly compared with controls （P〈0.05）. Conclusions RSM decreased P-ERK and P-JNK in activated HSCs, Which maybe one of important ways to block liver fibrosis.

关 键 词：肝纤维化 肝星状细胞 丹参 有丝分裂素激活蛋白激酶类 

分 类 号：R575.2[医药卫生—消化系统]