摘要
目的探讨早期应用血必净联合法舒地尔对脓毒症急性肾损伤后肾小管细胞凋亡的影响及机制。方法将60只雄性SD大鼠随机分为分假手术组、脓毒症组、血必净组、法舒地尔组和联合用药组,每组12只。以盲肠结扎穿孔法制造脓毒症模型,假手术组麻醉后开腹翻动肠道,随即关腹;脓毒症组按4ml/kg尾静脉注射0.9%氯化钠注射液;血必净组按4ml/kg注射血必净;法舒地尔组按20mg/kg注射盐酸法舒地尔;联合用药组予等剂量血必净和法舒地尔注射。在造模后6、24h再将每组随机分为2个亚组,每个亚组6只。检测各时间点大鼠的肾功能和炎症指标,观察肾小管病理改变、测定肾小管损伤评分,采用TUNEL法观察肾小管凋亡细胞,并计算凋亡指数(AI)。用免疫印迹法测定ROCK-1、MYPT-1和caspase-3表达的变化。结果(1)造模后24h时与脓毒症组相比,各用药组肾功能均能明显改善(P<0.05),在抑制TNF-α、IL-6表达上联合用药组优于两单药组(均P<0.05)。(2)造模后24h时联合用药组与两单药组比较,肾小管损伤评分和AI值明显降低(均P<0.05)。(3)造模后24h时联合用药组与两单药组比较,更能抑制MYPT-1的磷酸化,促使caspase-3表达下降(均P<0.05)。结论脓毒症早期血必净联合法舒地尔可以通过Rho/ROCK信号通路抑制凋亡蛋白的表达,减少肾小管细胞在脓毒症进程中的凋亡,发挥协同作用减轻脓毒症导致的急性肾损伤。
Objective To investigate the effect of Xuebijing combined with fasudil on tubular apoptosis in sepsis-induced acute kidney injure（SI-AKI） of rats.Methods Sixty male SD rats were randomly divided into five groups：sham group,sepsis group,Xuebijing group,fasudil group and combination group with 12 rats in each group.Sham operation was performed in rats of sham group;SI-AKI were induced by cecal ligation and puncture（CLP） surgery in other 4 groups.0.9% sodium chloride at4ml/kg,Xuebijing at 4ml/kg,fasudil at 20mg/kg and Xuebijing with fasudil were injected via caudal vein in sepsis group,Xuebijing group,fasudil group and combination group,respectively.Six rats were sacrificed in each group at 6h and 24 h after treatment,respectively.Renal function parameters,inflammation factors,pathological changes of tubules,renal tubular injury score were measured.The apoptosis of renal tubular cells was determined by TUNEL method,apoptotic indices（AI） were calculated.The expressions of ROCK-1,MYPT-1 and caspase-3 were detected by Western blot.Results At 24 h after treatment,the renal function in 3 treatment groups was significantly improved compared to sepsis group（P〈0.05）.The TNF-α,IL-6 levels in combination group were lower than those in other two treatment groups（all P〈0.05）.The tubular damage scores and AI values in combination group were more attenuated compared to other two treatment groups（all P〈0.05）.The phosphorylation of MYPT-1was more markedly inhibited and the caspase-3 expression was decreased in combination group compared to other two treatment groups（all P〈0.05）.Conclusion Early application of Xuebijing combined with fasudil can alleviate renal dysfunction and pathological changes,reduce the tubular cell apoptosis in SI-AKI rats,in which the Rho-kinase signaling pathway may be involved.
作者
孙雪东
严一核
刘芳
褚韦韦
张亦婷
应利君
SUN Xuedong YAN Yihe LIU Fang et al(Department of Critical Care Medicine, Shaoxing People＇s Hospital, Shaoxing 312000, China)
出处
《浙江医学》
CAS
2017年第3期165-169,I0003,共6页
Zhejiang Medical Journal
基金
绍兴市科技计划(2014B70066)
