中药复方狼疮方对MRL/LPR狼疮小鼠肾组织干扰素及自身免疫炎症损伤的作用观察  

Effects of lupus recipe on renal interferon and autoimmunity injury in MRL/LPR mice

作　　者：刘瑞华 黄旋 吴海珊 叶红坚[1] 詹翠霞 范瑾瑾[1] 李晓艳[1] 阳晓[1] LIU Ruihua;HUANG Xuan;WU Haishan(Department of Nephrology,the First Affiliated Hospital,Sun Yat-sen University,Key Laboratory of Nephrology,National Health Commission and Guangdong Province,Guangzhou,510080)

机构地区：[1]中山大学附属第一医院肾内科,卫健委和广东省重点实验室,广州510080

出　　处：《中国中西医结合肾病杂志》2022年第1期6-10,I0001,共6页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.81774069、82074170)。

摘　　要：目的:通过研究狼疮方对MRL/LPR狼疮小鼠肾组织干扰素(IFN)及自身免疫炎症损伤的影响,探讨其治疗狼疮肾炎的作用机制。方法:将12周龄的雌性MRL/LPR狼疮小鼠随机分成模型组和狼疮方组,6~8周龄的雌性C57BL/6小鼠作为正常对照组。正常组和模型组予生理盐水0.2 ml灌胃,狼疮方组予狼疮方颗粒溶液(3.9 g/kg)0.2 ml灌胃,每天1次,连续给药4周。检测各组小鼠24 h尿蛋白、肾脏病理改变。免疫荧光检测肾组织免疫球蛋白G(IgG)含量;ELISA检测血清抗dsDNA抗体、抗核抗体(ANA)以及白细胞介素(IL)-1β和IL-10水平;qPCR检测肾组织IFNα和IFNγmRNA水平变化;试剂盒检测肾组织Caspase3酶活性变化;Western Blot检测肾组织非经典自噬通路LC3及Rubicon蛋白表达。结果:与狼疮小鼠模型组比较,狼疮方治疗组小鼠第2周和第4周的24 h尿蛋白水平明显下降(2.58±1.30)g vs.(0.99±0.10)g;(3.26±1.92)g vs.(1.22±0.41) g,P均<0.05。血清抗dsDNA抗体、ANA抗体及IL-1β显著降低,IL-10与模型组比较无显著差别。肾小球细胞增生及间质炎症细胞浸润减轻,IgG沉积显著减少;肾组织IFNαmRNA表达显著降低,IFNγmRNA表达无显著变化;Caspase3活性显著下降(201.1±50.3%vs.128.9±49.4,P<0.05);LC3及Rubicon蛋白表达水平增加。结论:中药复方狼疮方能抑制狼疮小鼠肾组织IFNα表达,降低肾组织自身免疫炎症反应,显著改善狼疮小鼠肾脏损伤。Objective:We investigated the effects of Lupus Recipe on interferon(IFN) and renal autoimmunity injury in MRL/LPR mice, with the aim to explore the potential mechanism of Lupus Recipe treating lupus nephritis. Methods:Female MRL/LPR mice aged 12 weeks were classified into model group and Lupus Recipe treatment group. Female C57 BL/6 mice age 6-8 weeks were served as normal control group. The control and model group mice were given 0.2 ml normal saline solution intragastrically, while Lupus Recipe treatment group mice received 0.2 ml(3.9 g/kg) Lupus Recipe solution. The drugs were given once daily for 4 weeks. The 24-hour urine protein and renal pathological changes of mice were examined. Immunofluorescence was used to examine immunoglobulin G deposition in kidney of mice. ELISA was used to detect serum dsDNA, and antinuclear antibodies(ANA), as well as serum cytokines of IL-1β and IL-10 levels. QPCR was employed to determine the mRNA levels of IFN-α and IFNγ in renal tissue of mice in each group. The caspase3 enzyme activity kit was used to assess the enzyme activity of caspase3 in kidney of mice. Western Blot was used to determine the expression of noncanonical autophagy protein of LC3 and Rubicon. Results:Compared to the lupus mouse model, Lupus Recipe treatment group mice had significantly decreased levels of 24-hour proteinuria at week 2 and week 4(2.58±1.30)g vs.(0.99±0.10)g, P<0.05;(3.26±1.92)g vs.(1.22±0.41)g, both P<0.05, as well as the serum levels of dsDNA autoantibodies, ANA antibodies, and inflammatory cytokine IL-1β. The proliferation of renal cells, infiltration of inflammatory cells, IgG deposition, the expression level of IFNα mRNA and activities of Caspase3(201.1±50.3% vs. 128.9±49.4%, P<0.05) in kidney were reduced significantly, while protein expression of LC3 and Rubicon increased remarkably. Serum level of IL-10 and renal mRNA level of IFNγ were comparable between the model group and Lupus Recipe treatment group. Conclusion:Lupus Recipe treatment could inhibit renal IFNα and infl

关 键 词：狼疮肾炎 狼疮方 肾脏损伤 自身免疫炎症反应 

分 类 号：R285.5[医药卫生—中药学]