长托宁预处理激活Sirt1抑制失血性休克复苏大鼠肺组织ROS/TRAF6减轻肺损伤

Pretreatment with penehyclidine inhibits ROS/TRAF6 in lung tissue of hemorrhagic shock resuscitation rats by activating SIRT1 and alleviating lung injury

目的探讨长托宁对失血性休克复苏(hemorrhagic shock resuscitation,HSR)损伤模型大鼠肺损伤的保护作用及可能机制。方法 30只SD大鼠随机分为假手术组、HSR组以及长托宁复苏组,每组10只,通过股动脉放/回血造成HSR模型。评价各组大鼠血气分析指标和氧合指数;HE染色观测各组大鼠肺损伤情况;ELISA方法检测各组大鼠肺泡灌洗液中IL-1β、IL-6、SOD和MDA含量;检测各组大鼠损伤肺组织线粒体中ROS的表达;Western blot方法检测各组大鼠损伤组织中Sirt1、NF-κB p65、IRAK4、p-IRAK4和TRAF6蛋白水平的表达。结果 HSR可减弱大鼠换气功能,造成肺损伤,显著升高肺泡灌洗液中IL-1β、IL-6和MDA含量,降低SOD活性,促进损伤肺组织线粒体中产生ROS,并增加大鼠损伤肺组织中NF-κBp65、TRAF6蛋白表达量,降低Sirt1、p-IRAK4蛋白表达;长托宁能显著提高HSR大鼠换气功能,改善肺损伤,显著降低肺泡灌洗液IL-1β、IL-6和MDA含量并升高SOD活性,抑制损伤肺组织ROS产生,且显著降低大鼠肺组织中NF-κB p65、TRAF6蛋白表达量,同时增加Sirt1、p-IRAK4蛋白表达。结论长托宁预处理能够通过激活Sirt1,抑制ROS/TRAF6的表达减轻失血性休克复苏大鼠肺损伤。

Objective To investigate the protective effect of penehyclidine hydrochloride on lung injury in rats with hemorrhagic shock resuscitation(HSR) injury and its possible mechanism. Methods Thirty SD rats were randomly divided into sham group, HSR group and penehyclidine resuscitation group, with 10 rats in each group, and the HSR model was created by the femoral artery blood injection/return.To evaluate the blood gas analysis indexes and oxygenation index of each group of rats;The lung injury of each group of rats were observed by HE staining;The contents of IL-1β, IL-6, SOD and MDA in the alveolar lavage fluid of each group of rats were detected by ELISA;The expression of ROS in the mitochondria of injured lung tissue of rats in each group;The protein expression of Sirt1, NF-κB p65, IRAK4, p-IRAK4 and TRAF6 in injured tissues were detected by Western blot. Results HSR could weaken the ventilation function of rats, caused lung injury, significantly increased the content of IL-1β, IL-6 and MDA, reduced SOD activity in the bronchoalveolar lavage fluid(BALF), promoted the production of ROS in the mitochondria of damaged lung tissue, and increased the expression of NF-κB p65 and TRAF6 protein in lung tissues of rats, and reduced Sirt1 and p-IRAK4 protein expression;Penehyclidine could significantly improve the ventilation function of HSR rats, improved lung injury, and significantly reduced IL-1β, IL-6 and MDA content and increased SOD activity in BALF, inhibited the generation of ROS in injured lung tissues, and significantly reduced the expression of NF-κB p65 and TRAF6 protein in rat lung tissues, while increasing the expression of Sirt1 and p-IRAK4 protein. Conclusion Pretreatment with penehyclidine hydrochloride can reduce lung injury in rats with hemorrhagic shock and resuscitation by activating Sirt1 and inhibiting the expression of ROS/TRAF6.