七氟烷通过JAK2-STAT3通路对大鼠肝缺血再灌注损伤的影响  

Effect of sevoflurane via JAK2-STAT3 pathway on hepatic ischemia-reperfusion injury in rats

作　　者：马志勇[1] 李雪平[1] 侯振宇[1] 汪明强 吕金利[1] MA Zhi-yong;LI Xue-ping;HOU Zhen-yu;WANG Ming-qiang;LYU Jin-li(Department of General Surgery,No.153 Central Hospital of PLA,Zhengzhou Henan 450042,China)

机构地区：[1]解放军第153中心医院普外科,河南郑州450042

出　　处：《局解手术学杂志》2020年第9期693-697,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(30901575)。

摘　　要：目的探讨七氟烷在抗大鼠肝缺血再灌注损伤(HIRI)中的机制。方法健康雄性SD大鼠40只,随机分为假手术组(sham组)、模型组(HIRI组)、七氟烷干预组(SF组)、七氟烷联合AG490干预组(AG490组),每组10只。再灌注6 h后收集肝和血清标本。采用HE染色法观察大鼠肝形态;采用RT-PCR检测大鼠JAK2、STAT3 mRNA表达;采用Western blot检测JAK2、STAT3、p-JAK2、p-STAT3蛋白表达水平;采用酶联免疫吸附试验(ELISA)检测血清中谷丙转氨酶(ALT)、天门冬氨酸转氨酶(AST)、碱性磷酸酶(AKP)、白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)的水平;钴淬灭技术检测线粒体膜通透性转运孔(mPTP)开放程度。结果HE染色结果显示HIRI组肝小叶结构明显破坏,SF组小叶和汇管区炎症轻于HIRI组,AG490组肝细胞间有炎症浸润并出现明显水肿。HIRI组JAK2、p-JAK2、STAT3和p-STAT3蛋白水平均低于sham组(P<0.05),SF组各蛋白表达水平均高于HIRI组(P<0.05),而AG490组各蛋白表达水平均低于SF组(P<0.05)。各组的JAK2和STAT3 mRNA表达趋势与蛋白表达一致(P<0.05)。HIRI组ALT、AST、AKP、IL-1β、IL-6和TNF-α的水平均较sham组高(P<0.05),SF组各指标较HIRI组低(P<0.05),而AG490组较SF组高(P<0.05)。HIRI组mPTP开放程度较sham组高(P<0.05),SF组较HIRI组低,而AG490组较SF组高(P<0.05)。结论七氟烷可以明显改善大鼠HIRI,降低肝的免疫炎症反应,其机制可能与激活JAK2-STAT3通路进而抑制mPTP的过度开放有关。Objective To investigate the mechanism of sevoflurane in treatment of hepatic ischemia-reperfusion injury(HIRI)in rats.Methods A total of 40 healthy male SD rats were randomly divided into sham group,model group(HIRI group),sevoflurane intervention group(SF group),sevoflurane combined with AG490 intervention group(AG490 group),with 10 rats in each group.Liver and serum samples were collected after reperfusion for 6 hours.The liver morphology of rats was observed by HE staining method.The expression of JAK2 and STAT3 mRNA in rats was detected by RT-PCR.The expression levels of JAK2 protein,STAT3 protein,p-JAK2 protein and p-STAT3 protein were detected by western blot.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(AKP),interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Cobalt quenching technique was used to detect the degree of mitochondrial membrane permeability pore(mPTP)opening.Results HE staining results showed that the structure of hepatic lobule was destroyed obviously in HIRI group,the inflammation of lobule and portal area in SF group was slighter than that in HIRI group.There was inflammatory infiltration and obvious edema between hepatocytes in AG490 group.The levels of JAK2 protein,pJAK2 protein,STAT3 protein and pSTAT3 protein in HIRI group were lower than those in sham group(P<0.05),while the levels of those protein in SF group were higher than those in HIRI group(P<0.05),while the protein levels in AG490 group were lower than those in SF group(P<0.05).The trend of mRNA expression in each group was similar to the protein level(P<0.05).The levels of ALT,AST,AKP,IL-1β,IL-6 and TNF-αin HIRI group were higher than those in sham group(P<0.05),while the serum levels in SF group were lower than those in HIRI group(P<0.05),the serum levels in AG490 group were higher than those in SF group(P<0.05).The mPTP opening degree of HIRI group was higher than that of sham group(P<0.05),the mPTP o

关 键 词：七氟烷 肝缺血再灌注损伤 Janu激酶信号转导子 转录激活子 线粒体膜通透性转运孔 免疫炎症 

分 类 号：R971.2[医药卫生—药品]