艾司氯胺酮对大鼠气腹肠缺血再灌注损伤的作用及机制

Effect and Mechanism of Esketamine on Pneumoperitoneum Intestinal Ischemia-reperfusion Injury in Rats

目的:探讨艾司氯胺酮对气腹大鼠Elav样家族成员1(CELF1)/凋亡诱导因子(AIF)通路及肠缺血再灌注(I/R)损伤的影响。方法:100只20周龄Wistar大鼠分为正常对照组、模型组、丙泊酚组(200mg/ml)、艾司氯胺酮低剂量组(1000mg/ml)、高剂量组(2000mg/ml)。丙泊酚组、艾司氯胺酮低、高剂量组分别给予相应药物腹腔注射10min,正常对照组、模型组给予等体积生理盐水腹腔注射10min,然后模型组、丙泊酚组、艾司氯胺酮低、高剂量组大鼠进行气腹肠缺血再灌注120min。最终各组进行腹部撤回反射(AWR)评分评价大肠扩张度、取血清测定IL-2、TNF-α、DAO水平,处死大鼠测定肠湿重/干重比值、肠组织HE染色病理检查、对肠组织行CHiu评分、RT-PCR法及蛋白印记法测定肠组织中CUGBP信号通路的CELF1/AIF的mRNA和蛋白表达水平。结果:各组大鼠CHiu评分、肠湿重/干重比值、肠扩张度、血清IL-2、TNF-α、DAO水平、CELF1、AIF mRNA蛋白表达差异均有统计学意义(P均<0.05),模型组CHiu评分、肠湿重/干重比值、血清IL-2、TNF-α、DAO水平、CELF1、AIF mRNA和蛋白表达高于正常对照组、丙泊酚组、艾司氯胺酮各剂量组,肠扩张度低于正常对照组、丙泊酚组、艾司氯胺酮各剂量组(P均<0.05)。与艾司氯胺酮低剂量比较,艾司氯胺酮高剂量CHiu评分、肠湿重/干重比值、血清IL-2、TNF-α、DAO水平、CELF1、AIF mRNA和蛋白表达降低,肠扩张度升高(P均<0.05);艾司氯胺酮低剂量组CHiu评分、肠湿重/干重比值、血清IL-2、TNF-α、DAO水平、CELF1、AIF mRNA和蛋白表达高于丙泊酚组,肠扩张度低于丙泊酚组(P均<0.05);艾司氯胺酮高剂量组CHiu评分、湿重/干重比值、肠扩张度、血清IL-2、TNF-α、DAO水平、CELF1、AIF mRNA和蛋白表达与丙泊酚组比较差异无统计学意义(P>0.05)。结论:艾司氯胺酮对大鼠气腹肠I/R具有明显保护作用;其机制与艾司氯胺酮在肠I/R损伤中显著抑制CELF1/AIF通路有关。

Objective:To investigate the effect of esketamine on elav-like family member 1(CELF1)/apoptosis-inducing factor(AIF)pathway and intestinal ischemia-reperfusion injury in rats with pneumoperitoneum.Method:10020-week-old Wistar rats were divided into normal control group,model control group,propofol group(200mg/ml),low dose esketamine group(1000mg/ml)and high dose esketamine group(2000mg/ml).The propofol group,esketamine low and high dose groups were given intraperitoneal injections of the corresponding drugs for 10 minutes,and the normal control group and model control group were given intraperitoneal injections of an equal volume of normal saline for 10 minutes.Then the model control group,propofol group,esketamine low and high dose groups were subjected to pneumoperitoneum intestinal ischemia-reperfusion for 120minutes.Finally,the degree of large intestine dilatation of each group was evaluated by abdominal withdrawal reflex(AWR)score,and serum was collected to determine the levels of IL-2,TNF-α,and DAO.Intestinal tissues were subjected to CHiu score,and RT-PCR and Western blotting were used to determine the mRNA and protein expression levels of CELF1/AIF of the CUGBP signaling pathway in intestinal tissues.Results:There were statistically significant differences in CHiu score,intestinal wet weight/dry weight ratio,intestinal dilatation,serum IL-2,TNF-α,DAO levels,CELF1,AIF mRNA protein expression among the rats in each group(all P<0.05).The CHiu score,intestinal wet weight/dry weight ratio,serum IL-2,TNF-α,DAO levels,CELF1,AIF mRNA protein expression in the model group were higher than those in the normal control group,propofol group,and esketamine group,and the degree of intestinal distension lower than the normal control group,propofol group,and esketamine group(all P<0.05).Compared with low-dose esketamine,high-dose esketamine decreased CHiu score,intestinal wet weight/dry weight ratio,serum IL-2,TNF-α,DAO levels,CELF1,AIF mRNA protein expression,and increased intestinal distension(P<0.05);the CHiu score,intestinal wet weight/dry weight ratio,serum IL-2,TNF-α,DAO level,CELF1,AIF mRNA protein expression in the low-dose esketamine group were higher than those in the propofol group,and intestinal dilatation was decreased.The degree of CHiu score,wet weight/dry weight ratio,intestinal dilatation,serum IL-2,TNF-α,DAO levels,CELF1,AIF mRNA in the high-dose esketamine group were no significant difference compared with propofol group(P>0.05).Conclusion:Esketamine prevents intestinal ischemia-reperfusion injury in rats with pneumoperitoneum by inhibiting CELF1/AIF pathway.