摘要
目的:探讨抗纤软肝颗粒对PDGF诱导的肝星状细胞MEK-1和c-los表达的影响.方法:采用无血清培养,不同浓度的抗纤软肝颗粒温育肝星状细胞24h后,PDGF-BB(10pg/L)刺激24h,再加入上述浓度的抗纤软肝颗粒,3h后,又加入PDGF-BB(10pg/L)作用5min,然后收集细胞.采用MTT法测定细胞增生,免疫印迹化学发光法检测MEK-1,原位杂交法检测c-fosmRNA.结果:无血清培养显示抗纤软肝颗粒对于PDGF诱导的细胞增生具有抑制作用,并呈剂量依赖性,抗纤软肝颗粒5g/L、1.25g/L各组的MTT测定值分别为0.28±0.03和0.43±0.04,与PDGF对照组(0.82±0.05)相比P<0.01;对PDGF诱导的MEK-1及c-fosmRNA表达均有显著的抑制作用:抗纤软肝颗粒5g/L、1.25g/L各组细胞的MEK-1表达水平分别为0.143±0.013、0.169±0.007,与PDGF组(0.186±0.010)比较有显著陛差异(P<0.01);c-fosmRNA表达水平分别为0.152±0.010、0.163±0.005,与PDGF组(0.183±0.014)比较也显著减弱(P<0.01).结论:在所应用的剂量范围内,抗纤软肝颗粒可抑制PDGF诱导的HSC增生,其机制与干扰Ras-MEK-MAPK信号通路有关.
AIM: To investigate the effect of Kangxian ruangan keli (KXR) on the expression of MEK-1 and c-fos in hepatic stellate cell (HSC) indused by PDGF. METHODS: In a serum-free culture system, HSC was treated with a KXR preparation for 24 hours, followed by stimulation with PDGF-BB for 24 hours. Then the cells were incubated again in the medium containing KXR for 3 hours stimulated with PDGF-BB for 5 minutes, and collected. The proliferation of HSC was examined using an MTT assay. MEK-1 was detected with Western blotting and visualized by the enhenced chemiluminescent (ECL) method. The expression of c-fos mRNA was analyzed with in situ hybridization. RESULTS: The A values for the HSC growing in the media without and with addition of PDGF were 0.170±0.060 and 0.820±0.050, respectively. The PDGF-induced increase was hindered remarkably by KXR preparation in a dose-dependent manner. Reaction values for the systems with 5 g/L and 1.25 g/L of KXR were 0.280±0.030 and 0.430±0.040 respectively, lower significantly than that in the culture free of KXR (0.820±0.050, P <0.01). In addition, values for MEK-1 in HSC treated with 5 mg/mL and 1.25 mg/mL of KXR were 0.143±0.013, and 0.170±0.007, respectively, being lower than that in the cells treated only with PDGF-BB (0.186±0.010, P<0.01). The expression level of c-fos mRNA was 0.152±0.010 and 0.163±0.005, respectively, also lower than that of the PDGF group (0.183±0.014, P<0.01). CONCLUSION: Within the dose range used in the present study, KXR preparation shows an inhibitory effect on HSC proliferation induced by PDGF. The mechanism of this process may involve interference with Ras-MEK-MAPK singal transduction mediated by PDGF.
出处
《世界华人消化杂志》
CAS
2004年第2期347-350,共4页
World Chinese Journal of Digestology
基金
湖北省自然科学基金资助项目
No.2000J042湖北省教育厅科研基金资助项目
No.2000A06010~~
