CXCR1/CXCL8轴在原发性胆汁性胆管炎胆管上皮细胞异常增殖中的作用  

Roles of the CXCR1/CXCL8 axis in abnormal proliferation of bile duct epithelial cells in primary biliary cholangitis

作　　者：艾馨 付海艳 徐加敏 杨文霞 唐映梅[1] Ai Xin;Fu Haiyan;Xu Jiamin;Yang Wenxia;Tang Yingmei(Department of Gastroenterology,The Second Affiliated Hospital of Kunming Medical University,Kunming 650101,China)

机构地区：[1]昆明医科大学第二附属医院消化内科,昆明650101

出　　处：《中华肝脏病杂志》2023年第2期174-180,共7页Chinese Journal of Hepatology

基　　金：国家自然科学基金(81660102);云南省自然科学基金(2018FE001(-051))。

摘　　要：目的探讨CXC趋化因子受体1(CXCR1)/CXC趋化因子配体8(CXCL8)轴在原发性胆汁性胆管炎(PBC)胆管上皮细胞异常增殖中的作用。方法体内实验将30只雌性C57BL/6小鼠随机分为PBC模型组(PBC组)、Reparixin干预组(Rep组)、空白对照组(Con组),予以2-辛炔酸偶联牛血清白蛋白(2OA-BSA)联合聚肌苷酸聚胞苷酸(polyI:C)腹腔注射12周后建立PBC动物模型,成功建模后,Rep组予Reparixin皮下注射(2.5 mg·kg^(-1)·d^(-1),3周)。苏木精-伊红染色检测肝脏组织学变化,免疫组织化学法检测细胞角蛋白19(CK-19)表达,qRT-PCR检测肿瘤坏死因子-α(TNF-α)、γ干扰素(IFN-γ)和白细胞介素(IL)-6 mRNA表达,蛋白质印迹法(western blot)检测核转录因子-κB p65(NF-κB p65)、细胞外调节蛋白激酶1/2(ERK1/2)、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)表达。体外实验将人肝内胆管上皮细胞分为IL-8干预组(IL-8组)、IL-8+Reparixin干预组(Rep组)、空白对照组(Con组),IL-8组加入10 ng/ml人重组IL-8蛋白培养,Rep组加入10 ng/ml人重组IL-8蛋白培养后加入100 nmol/L Reparixin培养。EdU法检测细胞增殖情况,酶联免疫吸附法检测TNF-α、IFN-γ和IL-6表达,qRT-PCR检测CXCR1 mRNA表达,western blot检测NF-κB p65、ERK1/2及p-ERK1/2表达。数据组间比较采用单因素方差分析。结果体内实验结果显示,与Con组相比,PBC组小鼠肝脏内胆管上皮细胞增殖增多,NF-κB及ERK通路相关蛋白表达增高,炎症细胞因子表达增高,而经Reparixin干预后,上述结果被逆转(P<0.05)。体外实验表明,与Con组相比,IL-8组人肝内胆管上皮细胞增殖增多,CXCR1 mRNA表达增高,NF-κB及ERK通路相关蛋白表达增高,炎症细胞因子表达增高。与IL-8组相比,Rep组人肝内胆管上皮细胞增殖减少,NF-κB及ERK通路相关蛋白和炎症指标均显著降低(P<0.05)。结论CXCR1/CXCL8轴可调控PBC中胆管上皮�Objective To investigate the role of the CXC chemokine receptor 1(CXCR1)/CXC chemokine ligand 8(CXCL8)axis in the abnormal proliferation of bile duct epithelial cells in primary biliary cholangitis(PBC).Methods 30 female C57BL/6 mice were randomly divided into the PBC model group(PBC group),reparixin intervention group(Rep group),and blank control group(Con group)in an in vivo experiment.PBC animal models were established after 12 weeks of intraperitoneal injection of 2-octanoic acid coupled to bovine serum albumin(2OA-BSA)combined with polyinosinic acid polycytidylic acid(polyI:C).After successful modelling,reparixin was injected subcutaneously into the Rep group(2.5 mg·kg^(-1)·d^(-1),3 weeks).Hematoxylin-eosin staining was used to detect histological changes in the liver.An immunohistochemical method was used to detect the expression of cytokeratin 19(CK-19).Tumor necrosis factor-α(TNF-α),γ-interferon(IFN-γ)and interleukin(IL)-6 mRNA expression were detected by qRT-PCR.Western blot was used to detect nuclear transcription factor-κB p65(NF-κB p65),extracellularly regulated protein kinase 1/2(ERK1/2),phosphorylated extracellularly regulated protein kinase 1/2(p-ERK1/2),Bcl-2-related X protein(Bax),B lymphoma-2(Bcl-2),and cysteine proteinase-3(Caspase-3)expression.Human intrahepatic bile duct epithelial cells were divided into an IL-8 intervention group(IL-8 group),an IL-8+Reparicin intervention group(Rep group),and a blank control group(Con group)in an in vitro experiment.The IL-8 group was cultured with 10 ng/ml human recombinant IL-8 protein,and the Rep group was cultured with 10 ng/ml human recombinant IL-8 protein,followed by 100 nmol/L Reparicin.Cell proliferation was detected by the EdU method.The expression of TNF-α,IFN-γand IL-6 was detected by an enzyme-linked immunosorbent assay.The expression of CXCR1 mRNA was detected by qRT-PCR.The expression of NF-κB p65,ERK1/2 and p-ERK1/2 was detected by western blot.A one-way ANOVA was used for comparisons between data sets.Results The results of in vi

关 键 词：原发性胆汁性胆管炎 CXCR1 CXCL8 胆管上皮细胞 增殖 

分 类 号：R575.7[医药卫生—消化系统]