IL-18在红斑狼疮模型小鼠中的促炎作用及其在Wnt通路中的作用机制  

Proinflammatory effect of IL-18 in lupus erythematosus model mice and its mode of action in Wnt pathway

作　　者：殷洪 廷艳 曾家顺 YIN Hong;TING Yan;ZENG Jiashun(The Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Rheumatology and Immunology,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Nephrology,Guiqian International General Hospital,Guiyang 550018,Guizhou,China)

机构地区：[1]贵州医科大学附属医院,贵州贵阳550004 [2]贵州医科大学附属医院风湿免疫科,贵州贵阳550004 [3]贵黔国际总医院肾内科,贵州贵阳550018

出　　处：《贵州医科大学学报》2025年第4期520-528,共9页Journal of Guizhou Medical University

基　　金：贵阳市科技计划项目(筑科合同〔2019〕9-1-11);贵州省卫健委课题项目〔gzwkj2021-134〕;贵州医科大学附属医院国家自然科学基金培育项目(I-2020-24)。

摘　　要：目的评估白细胞介素-18(interleukin-18,IL-18)在红斑狼疮模型(MRL/lpr)小鼠中的促炎及肾脏损害作用,探讨其与Wnt/β-catenin信号通路中糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)、β-连环蛋白(β-catenin)蛋白的关系。方法选择5~6周龄雌性MRL/lpr小鼠分为模型组(n=8)及IL-18组(n=8),相同周龄、性别的C57BL/6小鼠为对照组(n=8);自8周龄起,对照组和模型组小鼠腹腔注射灭菌注射用水(0.1 mL/只),IL-18组腹腔注射IL-18注射液(500 ng/只),每天1次,持续8周;每两周检查检测1次各组小鼠尿蛋白水平;给药结束后麻醉处死小鼠,摘取小鼠脾脏、肾脏,计算脏器系数;ELISA法检测小鼠血清肌酐(serum creatinine,Scr)、抗双链DNA抗体(anti-double-stranded DNA antibody,Anti-dsDNA)、干扰素-γ(interferon-γ,IFN-γ)、IL-1β、IL-6及IL-10水平;HE染色观察肾脏组织结构改变及炎症浸润情况,IHC和Western blot检测肾组织中β-catenin、GSK-3β及pGSK-3β蛋白的表达情况。结果与对照组相比,模型组及IL-18组小鼠24 h尿蛋白水平在12~14周龄时开始升高,16周龄时IL-18组升高明显;与对照组比较,IL-18组肾脏、脾脏系数增大(P<0.05);肾脏HE染色结果显示,模型组及IL-18小鼠肾脏光镜下可见肾小球系膜细胞增生,炎症细胞浸润,肾小管变形坏死,以IL-18组小鼠病变最为显著;与对照组比较,IL-18组小鼠肾组织β-catenin、GSK-3β、p-GSK-3β蛋白表达增高(P<0.05);但GSK-3β蛋白在模型组和IL-18组的差异无统计学意义(P>0.05);与对照组比较,IL-18组Scr、Anti-dsDNA、IFN-γ水平升高(P<0.05);与对照组及模型组比较,IL-18组IL-1β、IL-6水平升高(P<0.05),IL-10水平降低(P<0.05)。结论IL-18在MRL/lpr小鼠中发挥促炎作用,其机制可能是通过抑制GSK-3β蛋白过度激活,阻止β-catenin蛋白降解而参与肾脏损害。Objective To evaluate the pro-inflammation and renal damaging effect of interleukin-18(IL-18)in mice with lupus erythematosus(MRL/lpr mice)and explore its relation to glycogen synthase kinase-3β(GSK-3β)andβ-catenin proteins in the Wnt/β-catenin signaling pathway.Methods Female MRL/lpr mice of 5-6 weeks of age were selected and divided into model group(n=8)and IL-18 group(n=8),with C57BL/6 mice of the same age and gender as control group(n=8).At 8 weeks of age,control and model groups were intraperitoneally injected(i.p.)with sterile water(0.1 mL/mouse),while IL-18 group was i.p.with IL-18(500 ng/mouse).The treatment was once a day for 8 weeks.Mouse urinary protein level in each group was measured biweekly.After the completion of administration,the mice were anesthetized and euthanized to collect spleens and kidneys for calculating organ indices.ELISA was used to examine mouse serum creatinine(Scr),anti-double-stranded DNA antibody(Anti-dsDNA),interferon-γ(IFN-γ),interleukin-1β(IL-1β),IL-6,and IL-10.HE staining was performed to observe the changes in renal tissue structure and inflammatory infiltration.IHC and Western blot were used to detect protein expressions ofβ-catenin,GSK-3β,and p-GSK-3β.Results When compared to control group,mouse 24 h urinary protein levels were increased in both model and IL-18 groups starting at 12-14 weeks of age,with significant elevation in IL-18 group by 16 weeks of age.IL-18 group exhibited higher kidney indices and spleen indices than control group did(P<0.05).HE staining results of the kidneys demonstrated that under the microscope,model and IL-18 groups displayed mesangial cell proliferation in glomeruli,inflammatory cell infiltration and tubular deformation and necrosis,with IL-18 group showing the most significant lesions.When compared to control group,IL-18 group showed increased expressions ofβ-catenin,GSK-3β,and p-GSK-3βproteins in mouse renal tissue(P<0.05).However,there was no statistically significant difference in GSK-3βprotein between model and IL-18 grou

关 键 词：白细胞介素-18 WNT GSK-3Β Β-CATENIN 系统性红斑狼疮 狼疮性肾炎 

分 类 号：R593.24[医药卫生—内科学]