CAJ | 학술논문

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24 h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1 ml,3h后更换2 ml完全培养液培养24 h.I/R组:更换缺血缓冲液1 ml,置于三气培养箱中3h模拟缺血,后更换2 ml完全培养液在正常条件下培养24 h模拟再灌注.OzoneOP组:在模拟缺血前,加入2 ml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.
목적 관찰재세포수평취양양화예처리(OzoneOP)대대서신소관상피세포재관주손상후적보호작용.방법 세포동보화24 h후분성3조,정상조、결혈재관주(I/R)조화OzoneOP조.정상조:경환대조완충액1 ml,3h후경환2 ml완전배양액배양24 h.I/R조:경환결혈완충액1 ml,치우삼기배양상중3h모의결혈,후경환2 ml완전배양액재정상조건하배양24 h모의재관주.OzoneOP조:재모의결혈전,가입2 ml취양화적배양액병작용2h,여동I/R조.채용류식세포의검측세포조망솔,병진행초양화물기화매(SOD)、병이철(MDA)적검측,Western blot검측B세포림파류/백혈병-2(bcl-2)、bcl-2상관X단백(bax)적표체.결과 정상조、I/R조화OzoneOP조적조망솔분별위(4.58±0.35)％、(32.16±3.27)％、(18.73±1.52)％,SOD활성분별위(14.31 ±1.31)、(7.03±3.94)、(10.45 ±2.35) U/mg단백,MDA함량분별위(0.48±0.12)、(2.13±1.44)、(1.36±0.59) nmol/mg단백.I/R조여정상조비교,bcl-2/bax비솔적강저(P＜0.05),이OzoneOP가이제고bcl-2/bax적표체비솔(P＜0.05).결론 취양양화예처리대신소관상피세포적I/R손상구유보호작용,기궤제가능통과조절bcl-2/bax적비솔래실현.
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 cultured 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/leukemia-2 (bcl-2) and bcl-2 associated X protein (bax) was detected by Western blotting.Results 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,L/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 bcl-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.