基于NF-κB信号通路探究狼疮定对红斑狼疮MRL/lpr小鼠炎症反应的调节作用  

Exploring the Regulatory Effect of Lupusidine on the Inflammatory Response of Lupus Erythematosus MRL/lpr Mice Based on NF-κB Signaling Pathway

作　　者：张芹 张丹君 曹莹 陈慕芝 杨科朋 ZHANG Qin;ZHANG Danjun;CAO Ying(The Third School of Clinical Medicine,Zhe-jiang Chinese Medical University,Hangzhou(310005),China;The Second School of Clinical Medicine,Zhejiang Chinese Medical University)

机构地区：[1]浙江中医药大学附属第三医院,杭州310005 [2]浙江中医药大学附属第二医院

出　　处：《浙江中医药大学学报》2023年第7期730-736,共7页Journal of Zhejiang Chinese Medical University

基　　金：浙江省中医药科研基金项目(2021ZB151);第七批全国老中医药专家学术经验继承工作项目(国中医药人教函〔2022〕76号)。

摘　　要：[目的]研究狼疮定对系统性红斑狼疮(systemic lupus erythematosus,SLE)MRL/lpr小鼠炎症反应的影响,并分析其对核因子-κB(nuclear factor-κB,NF-κB)信号通路的调节作用。[方法]将12只MRL/lpr小鼠随机分为模型组(6只)、干预组(6只),另选取C57BL/6小鼠6只为对照组。干预组灌胃给予0.1 mL/10 g狼疮定浓缩液,对照组、模型组给予等体积0.9%氯化钠溶液,所有动物均干预8周。给药前后分别测定血清炎性因子白细胞介素-6(interleukin-6,IL-6)、α-干扰素(interferon-α,IFN-α)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及NF-κB信号通路相关蛋白NF-κB抑制蛋白α(NF-κB inhibitorɑ,IκBα)、NF-κB抑制蛋白αp50(NF-κB inhibitorɑp50,IκBαp50)、NF-κB抑制蛋白αp65(NF-κB inhibitorɑp65,IκBαp65)表达水平,采用扩增阻碍突变系统-聚合酶链式反应(amplification refractory mutation system Real time fluorescence-quantitative polymerase chain reaction,ARMS-q PCR)检测NF-κB信号通路相关mRNA表达情况。通过苏木精-伊红染色检测肾组织损伤情况,采用免疫印迹法检测肾组织中炎性因子表达水平。[结果]与对照组比较,模型组血清炎性因子IL-6、IFN-α、TNF-α、NF-κB信号通路相关蛋白(IκBα、IκBαp50、IκBαp65)水平、NF-κB信号通路相关mRNA(IκBα、IκBαp50、IκBαp65)水平均显著升高(P<0.05),肾组织中IL-6、IFN-α、TNF-α表达水平升高(P<0.05)。模型组小鼠肾组织损伤明显,主要表现为肾小管扩张,肾小球硬化,结构严重破坏,且肾小球、肾小管周围出现明显的炎症反应。干预后,MRL/lpr小鼠血清各指标水平均出现明显降低(P<0.05),肾组织炎性损伤较模型组减轻。[结论]狼疮定可抑制MRL/lpr小鼠炎症反应,从而减轻炎性肾损伤,其作用机制可能与下调NF-κB信号通路有关。[Objective]To study the effect of lupusidine on the inflammatory response of MRL/lpr mice with systemic lupus erythematosus(SLE),and to analyze its regulatory effect on nuclear factor-κB(NF-κB)signaling pathway.[Methods]Twelve MRL/lpr mice were randomly divided into model group(6 mice)and intervention group(6 mice),and C57BL/6 mice(6 mice)were selected as control group.The intervention group was given 0.1 mL/10 g lupusidine concentrate by gavage,and the control group and model group were given an equal volume of 0.9%sodium chloride solution.All animals were intervened for 8 weeks.The expression levels of serum inflammatory factors interleukin-6(IL-6),interferon-α(IFN-α),tumor necrosis factor-α(TNF-α)and NF-κB signaling pathway-related proteins NF-κB inhibitorɑ(IκBα),NF⁃κB inhibitorɑp50(IκBαp50),NF⁃κB inhibitorɑp65(IκBαp65)levels were measured before and after administration.The expression of NF-κB signaling pathway-related mRNA was detected by amplification refractory mutation system Real time fluorescence-quantitative polymerase chain reaction(ARMS-qPCR).Hematoxylin-eosin(HE)staining was used to detect renal tissue damage,and Western blot was used to detect the expression levels of inflammatory factors in renal tissue.[Results]Compared with control group,the serum inflammatory factors IL-6,IFN-α,TNF-α,NF-κB signaling pathway related protein(IκBα,IκBαp50,IκBαp65)level,and NF-κB signaling pathway related mRNA(IκBα,IκBαp50,IκBαp65)levels in model group were significantly increased(P<0.05).The expression levels of IL-6,IFN-αand TNF-αin kidney tissue were increased(P<0.05).In model group,the kidney tissue was damaged significantly,mainly manifested as the expansion of renal tubules,glomerulosclerosis and severe structural damage,and obvious inflammatory reaction appeared around the glomerulus and tubules.After intervention,the serum levels of all indicators in MRL/lpr mice were significantly reduced(P<0.05).Inflammatory injury of kidney tissue was less than that of model grou

关 键 词：系统性红斑狼疮 MRL/LPR小鼠 NF-ΚB信号通路 狼疮定 炎症反应 肾损伤 

分 类 号：R331[医药卫生—人体生理学]