Taxol对缺氧培养乳鼠心肌细胞Cx43蛋白表达及分布的影响

中国临床药理学与治疗学 ›› 2013, Vol. 18 ›› Issue (9): 975-980.

Taxol对缺氧培养乳鼠心肌细胞Cx43蛋白表达及分布的影响

王德国¹, 王新¹, 王安才¹, 朱红军², 孙贤林²

¹皖南医学院附属弋矶山医院老年科,芜湖 241001,安徽;
²安徽省立医院心血管内科,合肥 231000,安徽

收稿日期:2013-02-18 修回日期:2013-08-05 发布日期:2013-09-07
作者简介:王德国,男,博士,讲师,副主任医师,研究方向:心律失常基础与临床研究。Tel: 0553-5739308 E-mail: wangdeguo@medmail.com.cn
基金资助:
国家自然科学基金(No81200142);安徽省自然科学基金(1208085QH156、1208085MH129);弋矶山医院人才引进科研基金(YR201001)

Effects of Taxol on the expression and distribution of connexin 43 in cultured cardiac myocytes under hypoxia

WANG De-guo¹, WANG Xin¹, WANG An-cai¹, ZHU Hong-jun², SUN Xian-lin²

¹Department of Gerontology, Yijishan Hospital of Wannan Medical College, Wuhu 241001, Anhui,China;
²Department of Cardiology, Anhui Provincial Hospital of Anhui Medical University, Hefei 231000, Anhui,China

Received:2013-02-18 Revised:2013-08-05 Published:2013-09-07

摘要/Abstract

摘要： 目的: 观察微管稳定剂Taxol对缺氧导致的培养乳鼠心肌细胞Cx43表达和分布的影响。方法: 差速贴壁梯度离心法分离培养乳大鼠心肌细胞,取培养 4 d 细胞缺氧 120 min,分别以不同浓度的Taxol干预;免疫印迹法检测聚合态微管蛋白含量、心肌细胞Cx43的蛋白表达,免疫荧光染色后激光共聚焦显微镜观察Cx43的分布。结果: 正常培养心肌细胞Cx43分布在核膜和细胞的闰盘处,缺氧 120 min 可导致心肌细胞微管解聚,Cx43蛋白表达降低,在心肌细胞间连接处分布规律散失,核膜Cx43分布减弱或消失,而均匀分布在细胞膜上;在低剂量的Taxol作用下,心肌细胞微管解聚状态缓解,Cx43蛋白表达和分布异常明显改善;随着Taxol剂量增加,这种改善作用更明显,呈现剂量依赖性。结论: 缺氧引起乳鼠心肌细胞微管解聚,Cx43蛋白表达降低分布紊乱,微管稳定剂Taxol可以显著地保护缺氧导致的心肌Cx43异常,具有潜在的抗缺血性心律失常价值。

关键词: 心肌, 缺氧, 微管, 缝隙连接, Taxol, 连接蛋白

Abstract: AIM: To observe the effects of microtubule stabilizer (Taxol) on the expression and distribution of Cx43 in cultured cardiac myocytes under hypoxia and investigate its new value as an anti-arrhythmic agent.METHODS: Cultured cardiac myocytes from newborn rats were exposed to hypoxia for 120 min with or without Taxol (a kind of microtubule stabilizer) in different concentrations. The expression of Cx43, polymerized tubulin and total tubulin were determined by western blots. Cx43 expression and distribution were detected by immunofluorescence staining and observed under laser scanning confocal microscope.RESULTS: In normal myocytes, Cx43 converge at nuclear membrane and intercalated discs in clusters. After 120 min hypoxia, microtubule in cardiac myocytes was depolymerized and Cx43 protein expression was reduced. Furthermore, Cx43 distribution was disorder which showing reduced at the intercalated disc and dispersively distributed in membranes. Pretreated with Taxol, microtubular depolymerization, Cx43 reduction and redistribution were inhibited in a dose dependent manner.CONCLUSION: Microtubular depolymerization and Cx43 redistribution induced by hypoxia were protected by low dosage of Taxol.Microtubule stabilizer may be acted as a new anti-arrhythmic agent.

Key words: Cardiac myocyte, Hypoxia, Microtubule, Gap junction, Taxol, Connexin

中图分类号:

王德国, 王新, 王安才, 朱红军, 孙贤林. Taxol对缺氧培养乳鼠心肌细胞Cx43蛋白表达及分布的影响[J]. 中国临床药理学与治疗学, 2013, 18(9): 975-980.

WANG De-guo, WANG Xin, WANG An-cai, ZHU Hong-jun, SUN Xian-lin. Effects of Taxol on the expression and distribution of connexin 43 in cultured cardiac myocytes under hypoxia[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(9): 975-980.

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