白细胞介素-23在急性肝损伤模型中调控Kupffer细胞M1型极化的作用机制  

IL-23 regulates M1 polarization of Kupffer cells in acute liver injury in vitro and in vivo

作　　者：华男 方莹 张芳芳 桑建忠[1] HUA Nan;FANG Ying;ZHANG Fangfang;SANG Jianzhong(Department of Gastroenterology,Yuyao People's Hospital,Ningbo 315400,China)

机构地区：[1]余姚市人民医院消化内科,315400

出　　处：《浙江医学》2021年第16期1729-1733,I0003,共6页Zhejiang Medical Journal

基　　金：浙江省医药卫生科技计划项目(2018KY756)。

摘　　要：目的探讨IL-23在急性肝损伤模型中调控Kupffer细胞M1型极化的作用机制。方法体外培养Kupffer细胞,用佛波酯(PMA)、脂多糖(LPS)和重组人IFN-γ诱导其M1型极化,将Kupffer细胞分为IL-23+anti-IL-23组、IL-23组、对照组;采用流式细胞术检测M1型细胞比例,采用ELISA法检测一氧化氮合酶(iNOS)、TNF-α、IL-6的水平,采用Western blot法检测JAK1、p-JAK1、STAT1及p-STAT1蛋白表达水平。将小鼠分为anti-IL-23组、IL-23组、模型组、对照组,采用四氯化碳诱导小鼠肝损伤;采用ELISA法检测小鼠肝脏组织中iNOS、TNF-α、IL-6的水平,免疫组织化学染色检测CD11c+的表达情况,HE染色观察小鼠肝脏病理改变,Western blot法检测小鼠肝脏组织中JAK1、p-JAK1、STAT1及p-STAT1蛋白表达水平。结果IL-23组中M1型细胞的比例为(12.05±1.32)%,而IL-23+anti-IL-23组中M1细胞的比例为(7.55±1.08)%,对照组中M1型细胞的比例为(8.32±1.32)%,IL-23组明显高于对照组(P<0.05)。IL-23组中iNOS、TNF-α、IL-6水平均明显高于对照组(均P<0.05),而IL-23+anti-IL-23组中iNOS、TNF-α、IL-6水平均明显低于IL-23组(均P<0.05)。IL-23组中JAK1、p-JAK1、STAT1以及p-STAT1蛋白表达水平均明显高于对照组(P<0.05);而IL-23+anti-IL-23组JAK1、p-JAK1、STAT1以及p-STAT1蛋白表达水平均明显低于IL-23组(P<0.05)。对照组小鼠iNOS、TNF-α、IL-6水平较其余3组均为低(均P<0.05);而模型组iNOS、TNF-α、IL-6水平较IL-23组、anti-IL-23组均为低(P<0.05)。对照组小鼠肝组织中未见CD11c+的表达,而模型组小鼠CD11c+表达明显,IL-23组小鼠CD11c+的表达较模型组增高。对照组小鼠肝组织无明显损伤或细胞炎症反应;而模型组小鼠肝组织出现明显的炎症反应和细胞损伤;IL-23组小鼠肝组织损伤较模型组明显;而anti-IL-23组小鼠肝组织损伤较IL-23组减轻。与对照组比较,IL-23组、模型组小鼠JAK1、p-JAK1、STAT1及p-STAT1蛋白表达水平较明显升高(均P<0.05);anti-IL-23�Objective To explore the mechanism of IL-23 promoting M1 polarization of Kupffer cells.Methods Kupffer cells were cultured in vitro,and their M1 polarization was induced by phorbol ester(PMA),lipopolysaccharide(LPS)and recombinant human IFN-γ.Kupffer cells were divided into control group,IL-23 group,IL-23+anti-IL-23 group.Flow cytometry was used to detect the ratio of F4/80+CD11c+cells,and enzyme-linked immunosorbent assay(ELISA)was used to detect the expression of iNOS,TNF-α,IL-6.Western blot was used to detect the expression of JAK1-STAT1.Acute liver injury was induced by carbon tetrachloride in mice,The mice were divided into control group,model group,IL-23 group and anti-IL-23 group.The expression of iNOS,TNF-αand IL-6 in peripheral blood was detected by ELISA,CD11c+in liver tissue and JAK1-STAT1 were detected by IHC and Western-blot.Results The proportion of M1 cells in the IL-23 group was(12.05±1.32)%,while the proportion of M1 cells in the IL-23+anti-IL-23 group was(7.55±1.08)%,and the proportion of M1 cells in the control group It was(8.32±1.32)%,the IL-23 group was significantly higher than the control group(P<0.05).The levels of iNOS,TNF-αand IL-6 in the IL-23 group were significantly higher than those in the control group(all P<0.05),while the levels of iNOS,TNF-αand IL-6 in the IL-23+anti-IL-23 group All were significantly lower than the IL-23 group(all P<0.05).The expression levels of JAK1,p-JAK1,STAT1,and p-STAT1 in the IL-23 group were significantly higher than those in the control group(P<0.05),while in the IL-23+anti-IL-23 group,JAK1,p-JAK1,STAT1,and The expression level of p-STAT1 protein was significantly lower than that of IL-23 group(P<0.05).The levels of iNOS,TNF-αand IL-6 in the control group were lower than those in the other three groups(all P<0.05),while the levels of iNOS,TNF-αand IL-6 in the model group were lower than those in the IL-23 group and anti-IL The-23 group was all low(P<0.05).There was no CD11c+expression in the liver tissue of mice in the control group,while the

关 键 词：白介素-23 急性肝损伤 KUPFFER细胞 M1极化 

分 类 号：R575[医药卫生—消化系统]