CAJ | 학술논문

目的评价热休克蛋白70(HSP70)和血红素加氧酶-1(HO-1)表达在肾缺血后处理减轻肾缺血再灌注损伤中的作用.方法健康雄性SD大鼠140只,体重250～280 g,采用随机数字表法,将大鼠随机分为4组(n=35):假手术组(S组)仅开腹,游离双侧肾脏,分离双侧肾蒂不夹团;肾缺血再灌注组(I/R组)夹闭双侧肾蒂缺血45 min,恢复灌注;缺血后处理组(IPo组)夹闭双侧肾蒂45 min,再灌注10 s,缺血10 s,反复3次,恢复灌注;HSP抑制剂槲皮黄酮+缺血后处理组(Q+IPo组)缺血前1 h 腹腔注射槲皮黄酮100 mg/kg,余操作同IPo组.于再灌注即刻(T0)、1、3、6、12、24、48 h(T1～6)时各组随机取5只大鼠抽心脏血后取肾,检测肾组织HSP70、HO-1的mRNA和蛋白表达,T3时抽心脏血,测定血清肌酐(Cr)和尿素氮(BUN)浓度、caspase-3 mRNA的表达,TUNNEL法检测肾组织凋亡细胞,计算凋亡指数(AI),光镜下观察肾组织病理学结果.结果与S组比较,其余组T3时血清Cr和BUN浓度和AJ升高,caspase-3 mRNA表达上调,各时点HSF70、BO-1的mRNA和蛋白表达上调(P＜0.05);与I/R组比较,IPo组T3时血清Cr和BUN浓度和AI降低,caspase-3 mRNA表达下调,T1～5时HSP70、HO-1的mRNA和蛋白表达上调(P＜0.05);与IPo组比较,Q+IPo组T3时血清Cr和BUN浓度和AJ升高,caspase-3mRNA表达上调,T1～5时HSP70、HO-1的mRNA和蛋白表达下调(P＜0.05).IPo组肾组织病理学损伤较I/R组减轻,Q+IPo组肾组织病理学损伤程度与I/R组相似.结论 HSP70和H0-1表达参与了肾缺血后处理减轻肾缺血再灌注损伤的过程.
목적 평개열휴극단백70(HSP70)화혈홍소가양매-1(HO-1)표체재신결혈후처리감경신결혈재관주손상중적작용.방법건강웅성SD대서140지,체중250～280 g,채용수궤수자표법,장대서수궤분위4조(n=35):가수술조(S조)부개복,유리쌍측신장,분리쌍측신체불협단;신결혈재관주조(I/R조)협폐쌍측신체결혈45 min,회복관주;결혈후처리조(IPo조)협폐쌍측신체45 min,재관주10 s,결혈10 s,반복3차,회복관주;HSP억제제곡피황동+결혈후처리조(Q+IPo조)결혈전1 h 복강주사곡피황동100 mg/kg,여조작동IPo조.우재관주즉각(T0)、1、3、6、12、24、48 h(T1～6)시각조수궤취5지대서추심장혈후취신,검측신조직HSP70、HO-1적mRNA화단백표체,T3시추심장혈,측정혈청기항(Cr)화뇨소담(BUN)농도、caspase-3 mRNA적표체,TUNNEL법검측신조직조망세포,계산조망지수(AI),광경하관찰신조직병이학결과.결과 여S조비교,기여조T3시혈청Cr화BUN농도화AJ승고,caspase-3 mRNA표체상조,각시점HSF70、BO-1적mRNA화단백표체상조(P＜0.05);여I/R조비교,IPo조T3시혈청Cr화BUN농도화AI강저,caspase-3 mRNA표체하조,T1～5시HSP70、HO-1적mRNA화단백표체상조(P＜0.05);여IPo조비교,Q+IPo조T3시혈청Cr화BUN농도화AJ승고,caspase-3mRNA표체상조,T1～5시HSP70、HO-1적mRNA화단백표체하조(P＜0.05).IPo조신조직병이학손상교I/R조감경,Q+IPo조신조직병이학손상정도여I/R조상사.결론 HSP70화H0-1표체삼여료신결혈후처리감경신결혈재관주손상적과정.
Objective To evaluate the role of the expression of heat shock protein 70 (HSP70) and heme oxygenase-1 (HO-1) in the reduction of renal ischemia-reperfusion (I/R) injury by ischemic postconditioning in tats.Methods One hundred and forty healthy male SD rats weighing 250-280 g were randomized into 4 groups ( n = 35 each) : sham operation group (S group) ; I/R group; ischemic postconditioning group (IPo group); quercetin (an inhibitor of HSP) + ischemic postconditioning group (Q + IPo group). Renal I/R was produced by clamping bilateral renal pedicels for 45 min followed by reperfusion. In group S, bilateral kidneys were only exposed through a midline incision but their- pedicels were not clamped. In IPo and Q + IPo groups, 45 min ischemia was followed by three 10 s episodes of ischemia at 10 s intervals for reperfusion and in addition intraperitoneal quercetin 100 mg/kg was injected at 1 h before ischemia in group Q + IPo. Blood samples from hearts were obtained at 0, 1, 3, 6, 12, 24 and 48 h of reperfusion (T0-6) and the rats were then sacrificed and kidneys removed to detect the expression of HSP70 and HO-1 mRNA and protein in renal tissues. The blood samples obtained at T3 were used to determine serum creatinine (Cr) and urea nitrogen (BUN) concentrations and the expression of caspase-3 mRNA . The apoptosis in the renal tissues was detected using TUNEL and apoptotic index ( AI) was calculated. Microscopic examination was performed with light microscope. Results Compared with group S, the serum Cr and BUN concentrations and AI were significantly increased at T3,the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T0-6 in the other groups (P ＜ 0.05) . Compared with group I/R, the serum Cr and BUN concentrations and AI were significantly decreased at T3, the expression of caspase-3 mRNA was down-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was up-regulated at T1-5 in group IPo ( P ＜ 0.05) . Compared with group IPo, the serum Cr and BUN concentrations and AI were significantly increased at T3, the expression of caspase-3 mRNA was up-regulated at T3, and the expression of HSP70 and HO-1 mRNA and protein was down-regulated at T1-5, in group Q + IPo ( P ＜ 0.05) . The microscopic examination showed that the renal I/R injury was significantly attenuated by ischemic postconditioning and the degree of injury in group IPo was similar to that in group I/R. Conclusion The expression of HSP70 and HO-1 is involved in the reduction of renal I/R injury by ischemic postconditioning in rats.