Mechanisms of chloride in anoxia-reoxygenation injury of cultured rat ventricular myocytes
Mechanisms of chloride in anoxia-reoxygenation injury of cultured rat ventricular myocytes
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发表时间：
2007-06
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通讯作者：
Jie Chen;Dan Liu;He-Ping Chen;Z. Liao;Z. Lai;M. He
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作者：
Jie Chen;Dan Liu;He-Ping Chen;Z. Liao;Z. Lai;M. He
目的研究氯离子在原代培养大鼠心室肌细胞缺氧复氧损伤中的作用及机制。方法在缺氧-复氧（A/R）损伤条件下，分别用Cl-/HCO 3-抑制剂4-乙酰基-4 ′-异硫氰酸二苯乙烯-2，2 ′-二磺酸（SITS）、Cl-通道阻断剂蒽-9-羧酸（9-AC）、Na+，K+-2Cl-抑制剂布美他尼或等摩尔葡萄糖酸盐替代Cl-处理原代培养的大鼠心室肌细胞。检测细胞活力、丙二醛（MDA）含量、细胞内钙离子（[Ca 2 +]i）、乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性及核因子κ-B（NF-κB）。结果与对照组相比，复氧后A/R组MDA含量、[Ca 2 +]i、NFκB活性明显升高，细胞活力、LDH、SOD、GSH-Px活性明显降低。9-AC组和布美他尼组细胞活力、MDA、LDH、SOD和GSH-Px含量与A/R组比较差异无统计学意义。与A/R组相比，去Cl-组和SITS组MDA含量、[Ca 2 +]i、NF-κB B活性明显降低，细胞活力、SOD、GSH-Px活性明显升高。结论A/R诱导的Cl-增加是通过阴离子交换刺激介导的，Cl-增加参与了复氧诱导的Ca 2+超载和NF-κB转位。
Aim To study the role and mechanisms of chloride in anoxia-reoxygenation injury of primary cultured rat ventricular myocytes. Methods Under conditions of anoxia- reoxygenation(A/R) injury, primary cultured rat ventricular myocytes were treated with Cl-/HCO3- inhibitor 4-acetanide-4′-isothiocya- natostilbene -2,2′-disulfonic acid (SITS), Cl- channel blocker Anthracene-9-carboxylic acid(9-AC),Na+,K+-2Cl- inhibitor bumetanide or replaced Cl- with equimolar gluconate, respectively. The cell viability and contents of malondialdehyde(MDA) and intracellular Ca2+([Ca2+]i),activity of Lactate dehydrogenase (LDH), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)and Nuclear factor kappa-B (NF-κB) were measured. Results After reoxygenation, contents of MDA and [Ca2+]i, activity of NFκB were significantly increased in the A/R group while the cell viability and activities of LDH, SOD, GSH-Px decreased compared with those of control group. The cell viability and contents of MDA, LDH, SOD and GSH-Px in 9-AC or bumetanide group had no significance compared with those of A/R group. In Cl--free group or SITS group, contents of MDA and [Ca2+]i , activity of NF-κB were noticeably lower than those of the A/R group while the cell viability and activities of SOD, GSH-Px significantly increased compared with those of A/R group. Conclusions The A/R induced Cl- increase is mediated by anion exchange stimulation, and Cl- increase takes part in the reoxygenation induced Ca2+ overload as well as NF-κB transposition.