PI3K/AKT通路和S1PR2在急性梗阻性胆管炎诱导大鼠全身炎症反应中的作用  

Roles of PI3K/AKT-S1PR2 pathway in systemic inflammatory response induced by acute obstructive cholangitis in rats

作　　者：余伍勇 张文锋[1] 程瑶 龚建平[1] YU Wuyong;ZHANG Wenfeng;CHENG Yao;GONG Jianping(Department of Hepatobiliary Surgery,The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,P.R.China;Department of General Surgery,Shizhu Branch,The Second Affiliated Hospital of Chongqing Medical University and Shizhu People’s Hospital,Chongqing 409100,P.R.China)

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆400010 [2]重庆医科大学附属第二医院石柱分院·重庆市石柱土家族自治县人民医院普外科,重庆409100

出　　处：《中国普外基础与临床杂志》2019年第11期1278-1283,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金项目（项目编号：81701950）;重庆市科卫联合医学科研项目（项目编号：2016MSXM2-01）

摘　　要：目的研究急性梗阻性胆管炎(AOC)大鼠模型中外周血单个核细胞(PBMCs)中磷脂酰肌醇-3-羟激酶(PI3K)/AKT通路和鞘氨醇-1-磷酸受体2(S1PR2)的激活情况以及其对大鼠全身炎症反应的影响。方法①体外实验:分离和培养清洁级大鼠PBMCs,然后将其分为磷酸盐缓冲溶液对照组、单独PI3K抑制剂LY294002处理组、脂多糖处理组和脂多糖+LY294002处理组4组,收集各组细胞的上清液和总蛋白,检测细胞上清液中炎性因子肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平以及细胞中PI3K、AKT磷酸化水平和S1PR2蛋白的变化。②体内实验:60只清洁级SD大鼠被随机分为假手术组、单独PI3K抑制剂LY294002处理组、AOC模型组及AOC模型+LY294002处理组4组,记录大鼠存活情况,检测大鼠血清中肝功能丙氨酸转氨酶(ALT)、门冬氨酸氨基转移酶(AST)和总胆红素(TBIL)水平及血清中TNF-α和IL-6的水平以及大鼠PBMCs中PI3K、AKT磷酸化水平和S1PR2蛋白的变化。结果①体外实验结果:脂多糖+LY294002处理组细胞上清液中TNF-α和IL-6水平均明显低于脂多糖处理组(P<0.050),PI3K、AKT磷酸化水平和S1PR2的蛋白水平亦明显低于脂多糖处理组(P<0.050)。②体内实验结果:AOC模型+LY294002处理组大鼠的生存率高于AOC模型组,血清中肝功能ALT、AST、TBIL和炎性因子TNF-α和IL-6水平均明显低于AOC模型组(P<0.050),PI3K和AKT磷酸化水平及S1PR2蛋白表达水平也明显低于AOC模型组(P<0.050)。结论抑制AOC模型大鼠中PBMCs中PI3K/AKT通路的活化,可降低S1PR2的表达,且能抑制AOC诱导的大鼠全身炎症反应。Objective To investigate activation of phosphatidylinositol 3 hydroxykinase(PI3K)/AKT pathway and sphingosine 1-phosphate receptor 2(S1PR2)in peripheral blood mononuclear cells(PBMCs)of acute obstructive cholangitis(AOC)rats and their effects on systemic inflammation in rats.Methods①In vitro experiment:The isolated PBMCs from the rats were divided into 4 groups:a control group,LY294002 treatment group,lipopolysaccharide(LPS)treatment group,and LPS+LY294002 treatment group.The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in the supernatant were detected and the phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs were detected.②In vivo experiment:The rats were randomly divided into four groups:a control group,LY294002 treatment group,AOC model group,and AOC+LY294002 treatment group.The survival rate of rats was recorded,the liver function(ALT,AST,and TBIL),TNF-α,and IL-6 levels in the serum were detected.The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs of the rats were detected.Results①The results of in vitro experiment:The levels of TNF-αand IL-6 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group(P<0.050).The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group(P<0.050).②The results of in vivo experiment:The survival rate of rats in the AOC+LY294002 treatment group was higher than those in the AOC group.The serum levels of ALT,AST,TBIL,TNF-α,and IL-6 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group(P<0.050).The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group(P<0.050).Conclusion Inhibition of activation of PI3K/AKT pathway in PBMCs can inhibit expression of S1PR2,then alleviate systemic inflammatory response ind

关 键 词：急性梗阻性胆管炎 外周血单个核细胞 磷脂酰肌醇-3-羟激酶 AKT 鞘氨醇-1-磷酸受体2 

分 类 号：R73[医药卫生—肿瘤]