CAJ | 학술논문

目的评价青藤碱对肾缺血再灌注大鼠肾小管上皮细胞凋亡的影响及其与c-Jun氨基末端激酶(JNK)信号通路的关系.方法健康成年雄性Wistar大鼠54只,6～8周龄,体重180 ～220 g,采用随机数字表法分为3组(n=18):假手术组(S组)、肾缺血再灌注组(I/R组)和青藤碱组(SIN组).I/R组和SIN组采用夹闭左侧肾蒂45 min,并于再灌注即刻切除右肾制备肾缺血再灌注损伤模型.SIN组于再灌注前30 min腹腔注射青藤碱60 mg/kg,S组和I/R组在同一时间腹腔注射等容量生理盐水.于再灌注0.5、6、24 h时心脏穿刺采集血标本,测定血清Cr和BUN浓度,取血后,即刻行左肾切除术,光镜下观察肾组织病理学结果.采用免疫组化法检测肾小管上皮细胞磷酸化JNK(p-JNK)和caspase-3的表达.采用TUNEL法检测肾小管上皮细胞凋亡率.结果与S组比较,I/R组和SIN组血清Cr和BUN浓度升高,肾小管上皮细胞p-JNK和caspase-3表达上调,细胞凋亡率升高(P＜0.05);与I/R组比较,SIN组血清Cr和BUN浓度降低,肾小管上皮细胞p-JNK和caspase-3表达下调,细胞凋亡率降低(P＜0.05).SIN组肾组织病理学损伤较I/R组减轻.结论青藤碱减轻大鼠肾缺血再灌注损伤的机制与抑制JNK信号通路的激活,从而减少肾小管上皮细胞凋亡有关.
목적 평개청등감대신결혈재관주대서신소관상피세포조망적영향급기여c-Jun안기말단격매(JNK)신호통로적관계.방법 건강성년웅성Wistar대서54지,6～8주령,체중180 ～220 g,채용수궤수자표법분위3조(n=18):가수술조(S조)、신결혈재관주조(I/R조)화청등감조(SIN조).I/R조화SIN조채용협폐좌측신체45 min,병우재관주즉각절제우신제비신결혈재관주손상모형.SIN조우재관주전30 min복강주사청등감60 mg/kg,S조화I/R조재동일시간복강주사등용량생리염수.우재관주0.5、6、24 h시심장천자채집혈표본,측정혈청Cr화BUN농도,취혈후,즉각행좌신절제술,광경하관찰신조직병이학결과.채용면역조화법검측신소관상피세포린산화JNK(p-JNK)화caspase-3적표체.채용TUNEL법검측신소관상피세포조망솔.결과 여S조비교,I/R조화SIN조혈청Cr화BUN농도승고,신소관상피세포p-JNK화caspase-3표체상조,세포조망솔승고(P＜0.05);여I/R조비교,SIN조혈청Cr화BUN농도강저,신소관상피세포p-JNK화caspase-3표체하조,세포조망솔강저(P＜0.05).SIN조신조직병이학손상교I/R조감경.결론 청등감감경대서신결혈재관주손상적궤제여억제JNK신호통로적격활,종이감소신소관상피세포조망유관.
Objective To evaluate the effect of sinomenine on apoptosis in renal tubular epithelial cells of rats subjected to renal ischemia-reperfusion (I/R), and the relationship with C-Jun N-terminal kinase (JNK) signaling pathway.Methods Fifty-four male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were randomly divided into 3 groups (n =18 each) using a random number table: sham operation group (group S), I/R group and sinomenine group (group SIN).Renal ischemia was induced by occlusion of the left renal pedicle for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion in anesthetized rats in I/R and SIN groups.In group SIN, sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion, while the equal volume of normal saline was given instead of sinomenine at the same time point in S and I/R groups.Six animals in each group were selected at 0.5, 6 and 24 h of reperfusion, blood samples were collected by cardiac puncture for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.Immediately after blood sampling, the left kidney was removed for examination of pathological changes in renal tissues (with light microscopes) and for determination of phosphorylated JNK (p-JNK) and caspase-3 expression (by immune-histochemistry) and apoptosis in renal tubular epithelial cells (by TUNEL).The apoptotic rate was calculated.Results Compared with group S, the serum Cr and BUN concentrations were significantly increased, the expression of p-JNK and caspase-3 was up-regulated, and the apoptotic rate was increased in I/R and SIN groups.Compared with group I/R, the serum Cr and BUN concentrations were significantly decreased, the expression of p-JNK and caspase-3 was down-regulated, and the apoptotic rate was decreased in group SIN.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activation of p-JNK signaling pathway and reduced apoptosis in renal tubular epithelial cells of rats.