中国病理生理杂志2012，Vol.28Issue(8)：1399-1404,6.DOI:10.3969/j.issn.1000-4718.2012.08.011
细胞外低渗诱导大鼠胚胎心肌细胞H9c2容积激活性氯电流和调节性细胞容积回缩
Hypotonic challenges induce volume-activated chloride currents and regulatory volume decrease in rat embryonic myocardial H9c2 cells
摘要
Abstract
AIM: To invesligale the volume — aclivaled chloride currents and regulalory volume decrease (RVD) induced by hypolonic challenges in ral embryonic myocardial cell line H9c2. METHODS: The technique of whole — cell patch — clamp was used Lo record the chloride currenls induced by hypolonic challenges and to clarify the properlies of the currenls in H9c2 cells. The changes of cell volume were observed by the technique of real — lime living cell imaging, and the roles of chloride channels in RVD were analyzed. RESULTS: A weak background currenl was recorded in H9c2 cells under isolonic condition. Extracellular application of 47% hypolonic solution rapidly aclivaled an oulward rectified currenl, which did not exhibit lime — and vollage — dependent inaclivation with the currenl density of (47.77 ± 3. 80) pA/pF at +80 mV and ( -33.36 ±2.80) pA/pF at -80 mV. The reversal potential was ( -9.02 ±0.61) mV, closed to the calculated equilibrium potential for Cl- ( —0. 9 mV). The currenl was volume — sensitive and was completely suppressed by 47% hyperlonic solution. In addilion, chloride channel blockers lamoxifen (20 |xmol/L) , 5 — nitro —2 — (3 — phenylpropylamino) benzoic acid (NPPB,100 μmol/L) and ATP (10 mmol/L) significantly inhibited the currenl wilh different inhibitory ratios. The phenomenon of RVD was also observed in H9c2 cells under the condition of perfusion wilh 47% hypolonic solution. The chloride channel blocker NPPB at concentration of 100 μmol/L completely inhibited the RVD process. CONCLUSION: The volume — aclivaled chloride channels, which are aclivaled by extracellular hypolonic chal-lenges, play an important role in the process of regulalory volume decrease in H9c2 cells.关键词
H9c2细胞/氯通道/膜片钳术/容积调节Key words
H9c2 cells/ Chloride channels/ Patch - clamp techniques/ Volume regulation分类
医药卫生引用本文复制引用
刘振锋,厉冰雪,滕双凤,罗海,林娜,黄维渊,朱林燕,王立伟,陈丽新..细胞外低渗诱导大鼠胚胎心肌细胞H9c2容积激活性氯电流和调节性细胞容积回缩[J].中国病理生理杂志,2012,28(8):1399-1404,6.基金项目
国家自然科学基金资助项目(No.30870567 （）
No.30871267 （）
No.90913020 （）
No.U0932004) （）
广东省自然科学基金资助项目(No.7005974) （No.7005974）
