肿瘤坏死因子α介导人脐动脉血管平滑肌细胞成骨样钙化

李振勇;倪兆慧;钱家麒;戴慧莉;顾乐怡;郭永平;孙明姝

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中华肾脏病杂志 ›› 2008, Vol. 24 ›› Issue (1) : 30-34.
基础研究

肿瘤坏死因子α介导人脐动脉血管平滑肌细胞成骨样钙化

  • 李振勇;倪兆慧;钱家麒;戴慧莉;顾乐怡;郭永平;孙明姝
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Ossific calcification of human umbilical artery vascular smooth muscle cells induced by TNF-α

  • LI Zhen-yong, NI Zhao-hui, QIAN Jia-qi, DAI Hui-li, GU Le-yi, GUO Yong-ping, SUN Ming-shu
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摘要

目的 观察肿瘤坏死因子α(TNF-α)对体外培养人脐动脉血管平滑肌细胞(hUASMC)骨特异性碱性磷酸酶(BAP)、骨桥蛋白(OPN)和骨形成蛋白2(BMP-2)等成骨样分化标志蛋白表达以及细胞外钙盐沉积的影响。 方法 植块贴壁法原代培养人hUASMC。予以TNF-α刺激,甲O-酚酞络合酮方法测定细胞外基质钙盐沉积;Von Kossa法观察钙化染色;实时定量PCR法测定血管平滑肌细胞BAP和OPN mRNA表达水平;免疫印迹法观察血管平滑肌细胞BAP、OPN和BMP-2蛋白表达水平。 结果 一定浓度范围内的TNF-α可促进hUASMC增殖;增加细胞外基质钙盐沉积。TNF-α 50 μg/L刺激可在第3天上调血管平滑肌细胞BAP和OPN mRNA表达水平(BAP mRNA:1.908±0.034比1.000±0.033,OPN mRNA: 3.600±0.073比1.000±0.079,均P < 0.05)。TNF-α 50 μg/L刺激可在第5天上调血管平滑肌细胞BAP、OPN和BMP-2蛋白表达水平(BAP蛋白:3.394±0.083比1.000±0.030,OPN蛋白: 1.967±0.134比1.000±0.070,BMP-2蛋白:2.745±0.289比1.000±0.208, 均P < 0.05)。 结论 一定浓度范围的TNF-α可刺激hUASMC增殖;介导细胞成骨样分化;增加细胞外基质钙盐沉积,从而参与血管钙化的发生发展。

Abstract

Objective To investigate the effect of TNF-α on the expression of bone alkaline phosphatase (BAP), osteopontin (OPN) and bone morphogenetic protein 2 (BMP-2), and the deposit of extracellular calcium in human umbilical artery smooth muscle cells (hUASMC). Method hUASMC were cultured primarily with tissue explants-attached method. The mRNA expression of BAP and OPN was determined by real time-PCR. The protein expression of BAP, OPN and BMP-2 was determined by Western blotting. The calcium deposit was detected by O-cresolphthalein complexone method. Results TNF-α promoted the proliferation of hUASMC and increased the calcium deposit in a time- and dose-dependent manner. The mRNA expression of BAP and OPN in hUASMC was up-regulated by TNF-α(50 μg/L) treatment at day 3(BAP mRNA 1.908±0.034 vs 1.000±0.033, OPN mRNA 3.600±0.073 vs 1.000±0.079, all P<0.05). The protein expression of BAP, OPN and BMP-2 in hUASMC was up-regulated by TNF-α(50 μg/L) treatment at day 5 (BAP protein 3.394±0.083 vs 1.000±0.030, OPN protein 1.967±0.134 vs 1.000±0.070, BMP-2 protein 2.745±0.289 vs 1.000±0.208, all P<0.05). Conclusion TNF-α of certain concentrations can promote the proliferation of hUASMC, induce the ossific calcification of hUASMC, increase the calcium deposit, and participate in the development of vascular calcium

关键词

肿瘤坏死因子α / 肌细胞 / 平滑肌 / 脐动脉 / 骨生成

Key words

Tumor necrosis factor α / Myocytes / smooth muscle / Umbilical artery / Osteogenesis

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李振勇;倪兆慧;钱家麒;戴慧莉;顾乐怡;郭永平;孙明姝. 肿瘤坏死因子α介导人脐动脉血管平滑肌细胞成骨样钙化[J]. 中华肾脏病杂志, 2008, 24(1): 30-34.
LI Zhen-yong;NI Zhao-hui;QIAN Jia-qi;DAI Hui-li;GU Le-yi;GUO Yong-ping;SUN Ming-shu. Ossific calcification of human umbilical artery vascular smooth muscle cells induced by TNF-α[J]. Chinese Journal of Nephrology, 2008, 24(1): 30-34.
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