芪黄健脾滋肾颗粒改善MRL/lpr小鼠肾脏损害机制研究  

Mechanism of Qihuang Jianpi Zishen Granule on Improving Renal Damage in MRL/lpr Mice

作　　者：尚双双 李明[1] 黄传兵[1] SHANG Shuang-shuang;LI Ming;HUANG Chuan-bing(Rheumatology Department,The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031)

机构地区：[1]安徽中医药大学第一附属医院风湿病科,合肥230031

出　　处：《中国中西医结合杂志》2025年第3期321-329,共9页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：安徽省高校协同创新项目(No.GXXT-2021-085);安徽省卫生健康科研项目重点项目(No.AHWJ2022a005);2022年度省级中医优势专科建设项目-风湿病科(No.皖中医药服务秘[2022]34号)。

摘　　要：目的基于长链非编码RNA生长停滞特异性转录因子5(IncRNA GAS5)/miR-21/软脂酰化磷蛋白(Sprouty1)轴,研究芪黄健脾滋肾颗粒(QJZ)治疗MRL/lpr小鼠肾脏损害的疗效及作用机制。方法选取10只C57BL/6雌性小鼠为对照组,40只MRL/lpr雌性小鼠随机分为模型组、泼尼松组、QJZ组、吗替麦考酚酯组,每组10只;泼尼松组予以泼尼松溶液灌胃,QJZ组予以泼尼松+QJZ溶液灌胃,吗替麦考酚酯组予以泼尼松+吗替麦考酚酯溶液灌胃,对照组及模型组予以等量生理盐水,干预8周。二喹啉甲酸检测尿蛋白定量;ELISA检测肾损害指标[肌酐(SCr)和尿素氮(BUN)]、细胞因子[白介素(IL)-1、IL-6和肿瘤相关因子(TNF-α)、免疫学指标[抗dsDNA抗体、C3、C4];HE、Masson染色、肾脏病理损伤进行活动性指数(AI)及慢性指数(CI)评分评价小鼠肾脏组织病变;RT-qPCR检测小鼠肾脏lncRNA GAS5、miR-21-5p、Sprouty1、细胞外信号调节激酶(ERK)1/2、环磷酸腺苷反应元件结合蛋白(CREB)基因的表达;Western Blot检测小鼠肾脏Sprouty1、ERK1/2、CREB蛋白的表达。结果病理结果显示,模型组小鼠肾脏出现大量炎症、不同程度纤维化、系膜区细胞过度增殖等病理改变,AI与CI评分升高(P<0.01)。与模型组比较,QJZ能改善小鼠肾脏病理病变,降低AI与CI评分(P<0.01);降低尿蛋白、SCr、BUN(P<0.01),降低抗dsDNA抗体,升高补体C3、C4,降低细胞因子IL-1、IL-6和TNF-α水平(P<0.01)。与对照组比较,模型小鼠肾脏中GAS5的表达降低,miR-21-5p的表达升高,Sprouty1的表达下降(P<0.01),ERK/CREB通路的磷酸化水平升高(P<0.01)。与模型组比较,QJZ能够上调GAS5、下调miR-21-5p、上调Sprouty1的表达(P<0.01),抑制ERK/CREB通路的磷酸化(P<0.01)。结论QJZ可能通过lncRNA GAS5/miR-21-5p/Sprouty1信号轴,抑制ERK/CREB通路磷酸化,抑制肾小球系膜细胞增殖,减少炎症因子产生,改善系统性红斑狼疮肾脏损害。Objective To investigate the efficacy and underlying mechanism of Qihuang Jianpi Zishen Granule(QJZ)in the treatment of renal damage in MRL/lpr mice,based on the lncRNA growth arrest-specific 5(lncRNA GAS5)/miR-21/Sprouty1 axis.Methods Ten female C57BL/6 mice comprised the control group.Forty female MRL/lpr mice were randomly assigned into four groups:model group,prednisone group,QJZ group,and mycophenolate mofetil group,with ten mice per group.The prednisone group received a prednisone solution,the QJZ group was administered with a combination of prednisone and(QJZ)solution,and the mycophenolate mofetil group was given a combination of prednisone and mycophenolate mofetil solution.Both the control group and the model group received equivalent volumes of normal saline.The intervention period lasted for 8 weeks.Urinary protein levels were quantitatively measured using the bicinchoninic aci(BCA)assay.Renal damage indicators,including serum creatinine(SCr)and blood urea nitrogen(BUN),as well as cytokines(IL-1,IL-6,and TNF-α),and immunological markers(anti-dsDNA antibody,C3,C4)were assessed using ELISA.Histological examination of kidney lesions was performed using HE and Masson staining,activity index(AI)and chronic index(CI).The expression levels of lncRNA GAS5,miR-21-5p,Sprouty1,extracellular signal-regulated kinase(ERK)1/2,and cAMP response binding protein(CREB)genes in mouse kidneys were determined by RT-qPCR.The protein expression levels of Sprouty1,ERK1/2,and CREB in mouse kidneys were analyzed using Western Blot.Results Pathological analysis revealed significant inflammation,varying degrees of fibrosis,and excessive proliferation of mesangial cells in the kidneys of the model group.Additionally,AI and CI scores were significantly elevated(P<0.01).Compared with model group,QJZ significantly ameliorated renal pathological changes and reduced AI and CI scores in lupus mice(P<0.01).QJZ treatment resulted in decreased levels of urinary protein,SCr and BUN(P<0.01).It also reduced anti-dsDNA antibody levels and incr

关 键 词：MRL/LPR小鼠 肾损害 肾小球系膜细胞增殖 芪黄健脾滋肾颗粒 中药复方 

分 类 号：R285.5[医药卫生—中药学]