牛磺酸对肾NRK-52E细胞缺氧/复氧损伤的保护及初步机制  

Preliminary study on mechanism of protective effect of taurine in NRK-52E cells induced by hypoxia/reoxygenation injury

作　　者：龚慧[1] 万慧芳[1] 王美凤[2] 涂硕[1] 余乐涵[1] 杨晓红[1] 万福生[1] 

机构地区：[1]南昌大学医学院生物化学与分子生物学教研室,江西南昌330006 [2]南昌市第一医院护理部,江西南昌330008

出　　处：《中国药理学通报》2011年第7期961-965,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30060029)

摘　　要：目的观察牛磺酸(Tau)对NRK-52E细胞缺氧/复氧(H/R)损伤的保护作用及初步机制。方法用NRK-52E细胞缺氧8 h复氧12 h培养建立了H/R模型,流式细胞仪检测凋亡率和胞内游离钙离子浓度,RT-PCR和Western blot检测GRP78、Caspase-12、Caspase-3 mRNA和蛋白表达。结果与对照组比较,H/R组细胞凋亡率上升,GRP78、Caspase-12及Caspase-3 mRNA和蛋白表达均明显增高(P<0.01),胞质游离钙离子浓度也升高(P<0.01);与H/R组比较,各浓度牛磺酸处理组的细胞Caspase-3活性和凋亡率明显下降,胞质游离钙和GRP78、Caspase-12及Caspase-3 mRNA和蛋白表达均有明显降低(P<0.01)。结论牛磺酸对NRK-52E细胞H/R损伤有较好的抑制作用,且呈一定的剂量依赖性,其机制可能是调节胞内钙稳态、抑制GRP78、Caspase-12及Caspase-3表达,减少了细胞凋亡。Aim To investigate the mechanism of protective effect of taurine in NRK-52E cells induced by hypoxia/reoxygenation （ H/R ） injury. Methods NRK-52E cells were exposed to hypoxia for 5 hours fol- lowed by reoxygenation for 12 hours. Cell count and cell survival rate were determined by trypan blue exclusion test, and renal tubular epithelial cells intracellular Ca^2＋ concentration and apoptotic rate were analyzed by Flow Cytometry Analyzer, respectively. RT-PCR and Western blot were used to detect the mRNA and protein level of GRP78, Caspase-12 and Caspase-3. Resuits Compared with control, in the H/R group, cell apoptosis was obviously increased, the mRNA and protein expressions of GRP78, Caspase-12 and Caspase-3 were significantly increased （ P 〈 0. 01） , the concen-tration of intracellular Ca2 ＋ increased significantly; compared with H/R group, cell apoptosis, the concentration of intracellular Ca^2＋ , and the mRNA and protein expressions of GRP78, Caspase-12 and Caspase-3 in H/R ＋ Tau group fell dramatically （ P 〈 0. 01 ）. Conclusions Taurine can improve renal tubular epithelial cells NRK-52E hypoxia/ reoxygenation damage in a dose-dependent manner. The mechanism is mainly to relieve the intracellular calcium overload and downregulate the expressions of GRP78, Caspase-12 and Caspase-3 gene, and reduce apoptosis.

关 键 词：缺血/再灌注损伤 牛磺酸 葡萄糖调节蛋白 胱冬蛋白酶 NRK-52E细胞 肾脏 

分 类 号：R-332[医药卫生] R322.61