CAJ | 학술논문

目的评价异氟醚预处理对大鼠肾缺血再灌注时细胞凋亡的影响.方法 SD雄性大鼠30只,体重300～ 320 g,12～ 14周龄,采用随机数字表法将其分为3组(n=10):假手术组(S组)、肾缺血再灌注组(I/R组)和异氟醚预处理+肾缺血再灌注组(Iso+ I/R组).采用右肾切除、左肾蒂夹闭30 min时恢复灌注的方法建立肾缺血再灌注损伤模型.Iso+I/R组在密闭麻醉箱中吸入1.5％异氟醚(O22 L/min)1 h,洗脱30 min后制备模型.其余两组吸入O22 L/min 1 h.于再灌注2h时采集腹主动脉血样3 ml,测定血清肌酐(Cr)、尿素氮(BUN)、胱抑素C(CysC)浓度;取左肾组织,采用TUNEL法检测细胞凋亡情况,计算凋亡指数(AI);采用实时荧光定量PCR法检测p53 mRNA、BaxmRNA和Bcl-2 mRNA表达水平,采用Western blot法检测Bax、Bcl-2、caspase-3表达水平,计算Bcl-2和Bax表达的比值.结果与S组比较,I/R组血清Cr、BUN、CysC浓度升高,Bcl-2和Bax及其mRNA表达上调,Bcl-2/Bax比值降低,caspase-3表达上调,AI增加(P＜0.05);与I/R组比较,Iso+I/R组血清Cr、BUN、CysC浓度降低,Bcl-2 mRNA、Bax mRNA和Bax表达下调,Bcl-2/Bax比值升高,caspase-3表达下调,AI降低(P＜0.05);各组间p53 mRNA表达比较差异无统计学意义(P＞0.05).结论异氟醚预处理减轻大鼠肾缺血再灌注损伤的机制与其调节Bax和Bcl-2平衡、抑制肾细胞凋亡有关.
목적 평개이불미예처리대대서신결혈재관주시세포조망적영향.방법 SD웅성대서30지,체중300～ 320 g,12～ 14주령,채용수궤수자표법장기분위3조(n=10):가수술조(S조)、신결혈재관주조(I/R조)화이불미예처리+신결혈재관주조(Iso+ I/R조).채용우신절제、좌신체협폐30 min시회복관주적방법건립신결혈재관주손상모형.Iso+I/R조재밀폐마취상중흡입1.5％이불미(O22 L/min)1 h,세탈30 min후제비모형.기여량조흡입O22 L/min 1 h.우재관주2h시채집복주동맥혈양3 ml,측정혈청기항(Cr)、뇨소담(BUN)、광억소C(CysC)농도;취좌신조직,채용TUNEL법검측세포조망정황,계산조망지수(AI);채용실시형광정량PCR법검측p53 mRNA、BaxmRNA화Bcl-2 mRNA표체수평,채용Western blot법검측Bax、Bcl-2、caspase-3표체수평,계산Bcl-2화Bax표체적비치.결과 여S조비교,I/R조혈청Cr、BUN、CysC농도승고,Bcl-2화Bax급기mRNA표체상조,Bcl-2/Bax비치강저,caspase-3표체상조,AI증가(P＜0.05);여I/R조비교,Iso+I/R조혈청Cr、BUN、CysC농도강저,Bcl-2 mRNA、Bax mRNA화Bax표체하조,Bcl-2/Bax비치승고,caspase-3표체하조,AI강저(P＜0.05);각조간p53 mRNA표체비교차이무통계학의의(P＞0.05).결론 이불미예처리감경대서신결혈재관주손상적궤제여기조절Bax화Bcl-2평형、억제신세포조망유관.
Objective To evaluate the effect of isoflurane preconditioning on cell apoptosis during renal ischemia-reperfusion (I/R) in rats.Methods Thirty male Sprague-Dawley rats, aged 12-14 weeks, weighing 300-320 g, were randomly divided into 3 groups (n =10 each) using a random number table: sham operation group (group S);I/R group;isoflurane preconditioning plus I/R group (group Iso+I/R).The rats were anesthetized with intraperitoneal pentobarbital sodium 30 mg/kg.To establish the model of renal I/R injury, the right kidney was removed, and the left renal pedicle was occluded for 30 min with atraumatic mini-clamp for 30 min, followed by 2 h reperfusion.In Iso+I/R group, 1.5％ isoflurane was inhaled for 1 h, followed by 30 min of washout before I/R.In S and I/R groups only oxygen 2 L/min was inhaled for 1 h.Arterial blood samples were taken at 2 h of reperfusion to determine the concentrations of serum creatinine (Cr), blood urea nitrogen (BUN) and cystatin C (CysC).The animals were then sacrificed, and left kidneys were sampled for determination of the cell apoptosis (by TUNEL), expression of p53, Bax and Bcl-2 mRNA (using real-time fluorescent quantitative PCR), and expression of Bax, Bcl-2, and caspase-3 (by Western blot analysis).The Bcl-2/Bax ratio was calculated.Results Compared with group S, the serum Cr, BUN, and CysC concentrations were significantly increased, the Bcl-2/Bax ratio was decreased, Bcl-2 mRNA, Bax mRNA, Bcl-2, Bax and caspase-3 expression was up-regulated, and AI was increased in group I/R.Compared with group I/R, the serum Cr, BUN, and CysC concentrations were significantly decreased, the Bcl-2/Bax ratio was increased, Bcl-2 mRNA, Bax mRNA, Bax and caspase-3 expression was down-regulated, and AI was decreased in group Iso+I/R.There was no significant difference in p53 mRNA expression among the three groups.Conclusion Regulating the balance between Bcl-2 and Bax and inhibiting apoptosis in kidney cells are involved in the mechanism by which isoflurane preconditioning reduces renal I/R injury in rats.