摘要
目的:探讨中药复方"通脉口服液"对IL-1β诱导肾小球系膜细胞ERK通路活化的影响,进一步揭示中药复方"通脉口服液"抑制系膜细胞增生、延缓肾纤维化的可能机制。方法:应用细胞培养技术,进行肾小球系膜细胞的培养,采用血清药理学方法,制备中药复方"通脉口服液"、黄芪、三七药物血清,分别用IL-1β(终浓度1μg/L)刺激系膜细胞5、15、30min后,Western印迹法测定细胞外调节激酶(ERK)1/2的表达。分别加入10%中药复方"通脉口服液"、10%黄芪药物血清、10%三七药物血清以及特异性络氨酸激酶抑制剂genistein(10μg/mL)预处理12h后,观察磷酸化细胞外调节激酶(ERK)1/2的表达情况。结果:IL-1β刺激下,ERK活化在5min时出现高峰,后逐渐减弱,genistein预处理12h后,ERK活化延迟到30min时才出现。通脉、黄芪含药血清预处理12h后均可使IL-1 beta刺激下ERK活化延迟到15min时出现,30min时增强,且峰值较前减弱。三七含药血清预处理12h后在3个观察时间点上(5min、15min、30min)均未出现磷酸化。通脉、黄芪、三七含药血清与genistein联合使用后在3个观察时间点上(5min、15min、30min)均未观察到ERK的磷酸化。结论:中药复方"通脉口服液"、黄芪、三七含药血清对ERK的磷酸化均有一定的阻断作用,与genistein合用可增强这一阻断作用。组间相比较结果显示三七组作用要优于黄芪组,提示在抑制ERK的磷酸化作用方面,活血类中药作用可能要优于益气类中药;中药复方"通脉口服液"抑制系膜细胞增生、延缓肾纤维化的可能机制为抑制ERK抗体的磷酸化。
Objective ：To explore the inhibitory effect of TongMai Oral Solution （TM） on the expression of ERK1/2 MAPK signaling pathway. Methods ： Mesangial cell （MsC） were incubated with TM pharmacological serum,Milkvetch Root serum and Sanchi serum for 12h, genistein was added to the MsC separately in the presence of TM pharmacological serum for 12h, interleukin - 1β （IL - 113） for 0,5,15,30 min, respectively. Activities of ERK1/2 were assessed by Western bolt analysis. Results ： IL -1β can increased protein throsine phosphoralation, and the peak activation of ERK1/2 appearanced after 5 minutes. Use the inhibitor genistein of MAPKs pretreatment 12h,the appearance of the peak activation of ERK1/2 was delaved, After the TM serum,the Milkvetch Root serum or the Sanchl serum pretreatment 12h,the appearance of the peak activation of ERK1/2 was also delaved, especially at the group of Sanchi serum. When the genistein combine with the TM serum, the Miklvetch Root serum or the Sanchi serum pretreatment 12h,there were no expression of activation of ERK1/2. Conclusions ： TM pharmacological serum inhibited the expression of phospho - ERK1/2, this inhibited expression was enhanced by genistein. TM - mediated transduction possibly by signaling moleclrle ERK1/2 can suppress the cultured mesangial cell growth.
出处
《辽宁中医杂志》
CAS
北大核心
2007年第8期1053-1056,共4页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(30371836)
关键词
中药复方
通脉口服液
丝裂原活化蛋白激酶
ERK
系膜细胞
complex prescription of the traditional Chinese medicine
Tongmai Oral Solution
mitogen activated protein kinase
ERK
mesangial cell
