臭氧氧化预处理对体外大鼠肾小管上皮细胞缺血再灌注损伤的保护作用  

Protective effects of ozone oxidative preconditioning on NRK-52E against ischemia and reperfusion injury in vitro

作　　者：王磊[1] 刘修恒[1] 陈志远[1] 陈晖[1] 翁小东[1] 邱涛[1] 刘林[1] 

机构地区：[1]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2014年第6期1207-1209,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年基金资助项目（30901494）

摘　　要：目的观察在细胞水平臭氧氧化预处理（OzoneOP）对大鼠肾小管上皮细胞再灌注损伤后的保护作用。方法细胞同步化24h后分成3组，正常组、缺血再灌注（I／R）组和OzoneOP组。正常组：更换对照缓冲液1ml，3h后更换2m1完全培养液培养24h。I／R组：更换缺血缓冲液1ml，置于三气培养箱中3h模拟缺血，后更换2ml完全培养液在正常条件下培养24h模拟再灌注。OzoneOP组：在模拟缺血前，加入2ml臭氧化的培养液并作用2h，余同I／R组。采用流式细胞仪检测细胞凋亡率，并进行超氧化物歧化酶（SOD）、丙二醛（MDA）的检测，Western blot检测B细胞淋巴瘤／白血病-2（bcl-2）、bcl-2相关X蛋白（bax）的表达。结果正常组、I／R组和OzoneOP组的凋亡率分别为（4．58±0．35）％、（32．16±3．27）％、（18．73±1．52）％，SOD活性分别为（14．31±1．31）、（7．03±3．94）、（10．45±2．35）U／mg蛋白，MDA含量分别为（0．48±0．12）、（2．13±1．44）、（1．36±0．59）nmol／mg蛋白。I／R组与正常组比较，bcl-2／bax比率的降低（P〈0．05），而OzoneOP可以提高bcl-2／bax的表达比率（P〈0．05）。结论臭氧氧化预处理对肾小管上皮细胞的I／R损伤具有保护作用，其机制可能通过调节bcl-2／bax的比率来实现。Objective To investigate the effects of ozone oxidative preconditioning （OzoneOP） on NRK-52E cells against ischemia and reperfusion （I/R） injury. Methods After all the cells were synchronized for 24 h, they were divided into three groups： the normal group, I/R group and OzoneOP group. In normal group, control buffer （ 1 ml） was added, and 3 h later, 2 ml complete medium was given for culture of 24 h. In I/R group, the ischemic buffer （1 ml） was added and the cells were cuhured under ischemic condition for 3 h, then 2 ml complete medium was given for culture of 24 h. In OzoneOP group, before the simulated ischemia, 2 ml ozonated medium was given and cells were incubated for 2 h, and the rest were the same as I/R group. Flow cytometry was performed to detect cell apoptosis rate. The activity of superoxide dismutase （SOD） and the content of malondialdehyde （MDA） were tested. The expression of B cell lymphoma/lenkemia-2 （bel-2） and bcl-2 associated X protein （bax） was detected by Western blotting. Restilts The cell apoptosis rate in normal group, I/R group and OzoneOP group was （4. 58 ± 0. 35 ） % , （32. 16 ± 3.27 ）% and （18.73± 1.52）% , respectively. The SOD activities in normal group, I/R group and OzoneOP group were （ 14. 31 ± 1.31 ）, （ 7. 03 ± 3. 94） and （ 10. 45 ± 2. 35） U/mg protein, respectively. The MDA contents in normal group, I/R group and OzoneOP group were （0.48 ±0. 12）, （2. 13 ± 1.44） and （ 1.36 ± 0. 59） nmol/mg protein, respectively. The ratio of bel-2/bax expression was downregulated in I/R group as compared with normal group. And OzoneOP could increase this ratio. Conclusion OzoneOP has protective effects on NRK-52E against I/R injury probably by regulating the ratio of the bcl-2/bax expression.

关 键 词：臭氧氧化预处理 缺血再灌注损伤 肾小管上皮细胞 

分 类 号：R692.5[医药卫生—泌尿科学]