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机构地区:[1]北京医科大学第一医院肾内科肾脏病研究所,北京100034
出 处:《基础医学与临床》2000年第1期41-44,共4页Basic and Clinical Medicine
基 金:国家教委博士点基金! 9802;跨世纪人才基金! 39910210474-231-C04
摘 要:为观察低密度脂蛋白(LDL)对肾上管上皮细胞的影响及涉及的相关信号传导机制,以人近曲行小管上皮细胞系(HK-2)为对象,采用3H-TdR掺入与细胞计数测定增殖;特异性底物磷酸化法测定细胞外信号调节激酶(Eng)活性;凝胶迁移率法检测转录活化因于(AP-1)DNA结合活性。发现LDL100~500ug/mL刺激HK-2细胞72h可促进其增殖:加入LDL250ug/mL后2~60min内可使该细胞ERK活性明显增加,10min时达高峰;不同剂量(50~500ug/mL)刺激时,ERK活性分别较对照组增加1.56~2.19倍。LDL促使核内转录因子AP-1结合活性增加。EIUL特异性阻断剂PD(50umol/L)可以分别阻断LDL刺激的HK-2细胞ERK活性和DNA合成。RMA阻断PKC信号途径后,LDL对于ERK的活化效应明显减弱。In order to study the effect of low density lipoprotein(LDL)on the proliferation of human proximal tubular cells(HK-2) and its intercellular signal transduction mechanism, cell proliferation was assayed with 'H-TdR incorporation methods and cell counting. The activity ofextracellular signal regulated-protein kinase(Eax)was determined by specific substrate phosphorylation assay. Electrophoresis-mobility shift assay(EMSA) was used to detect the activity oftranscriptional complex AP-1. LDL increased 3H-TdR incorporation and cell number at concentration of 100mg/mL to 500mg/mL, the activity of Ear was stimulated by 250mg/mL of LDL from 2min. to 60min, and peaked at 10min. The induction of ERK activity by LDL was 1.56 ~2. 19 fold when compared with control. LDL also enhanced the activity of AP-1 in HK-2 cells. The inhibitor of ERK, PD blocked both proliferation and ERK activity of HK-2 cells. PMA, the inhibitor of protein kinase C, blocked the activity of Ear induced by LDL. It was concluded that LDL induced proliferation of HK-2 cells associated with ERK/AP-1 signal transduction pathway. Additionally, the activation of PKC may be implicated in the process.
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