麻黄连翘赤小豆汤及其效应成分抑制IgA肾病大鼠替代途径补体激活的研究  

Mahuang Lianqiao Chixiaodou Decoction and its active components inhibit alternative pathway complement activation in rat model of IgA nephropathy

作　　者：宋婷 盛广宇[1,2,3,4] 阮伟 张亚亨 杨雪军 SONG Ting;SHENG Guang-yuu;RUAN Wei;ZHANG Ya-heng;YANG Xue-jun(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Traditional Chinese Medicine Nephrology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Key Laboratory of Hepatorenal Diseases of Ministry of Education,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海中医药大学中医肾病研究所,上海201203 [3]上海中医药大学肝肾疾病病证教育部重点实验室,上海201203 [4]上海市中医临床重点实验室,上海201203

出　　处：《中国中药杂志》2025年第6期1626-1636,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82274492);上海市科委基金资助项目(22Y31920200)。

摘　　要：探究麻黄连翘赤小豆汤(MHLQ)及其主要效应成分治疗IgA肾病(IgAN)的可能作用机制。采用牛血清白蛋白(BSA)灌胃、皮下注射四氯化碳(CCl4)、尾静脉注射脂多糖(LPS)等综合措施建立IgAN大鼠模型。成模后,将IgAN大鼠随机分为模型组,MHLQ低、中、高剂量组,连翘苷(PHI)组,盐酸伪麻黄碱(PSE)组及氯沙坦钾(LP)组,并以Wistar大鼠为对照。MHLQ低、中、高剂量组分别给予MHLQ颗粒剂1.773、3.545、7.090 g·kg^(-1)灌胃,PHI组给予PHI 0.020 g·kg^(-1)灌胃,PSE组给予PSE 0.020 g·kg^(-1)灌胃,LP组给予LP 9.003 mg·kg^(-1)灌胃,对照组和模型组每日灌胃等体积生理盐水,各组均连续干预4周。给药结束后收集各组大鼠尿液、血清、肝组织和肾组织。检测24 h尿蛋白定量(24 h UTP)和肾功能;病理染色观察肾组织病理改变;免疫荧光(IF)检测肾脏IgA和补体C3/C3b/C3c表达;电镜观察肾组织超微结构;酶联免疫吸附测定(ELISA)法检测血清和肾组织补体C3、可溶性端补体复合物(sC5b-9)蛋白表达;蛋白免疫印迹(Western blot)法检测肝、肾补体C3/C3b/C3c、C5/C5a、C5b-9及B因子(CFB)蛋白表达;免疫组化(IHC)进一步检测肾组织补体C3蛋白表达。结果显示,与对照组比较,模型组大鼠尿素氮(BUN)、血肌酐(SCr)含量升高,肾小球代偿性扩张,球内系膜细胞增殖,系膜基质扩张,肾小球可见IgA免疫复合物或电子致密物沉积;血清补体C3含量增加;肾组织CFB,补体C3/C3b/C3c、C5/C5a、C5b-9蛋白表达升高;肝组织中补体C3/C3b/C3c和C5b-9蛋白表达升高。而经过MHLQ及其效应成分干预后,上述指标皆逆转。综上所述,MHLQ及其效应成分干预IgAN大鼠疗效肯定,能够改善肾功能,减轻肾脏免疫复合物沉积和病理损伤,其机制可能与抑制IgAN大鼠替代途径(AP)补体激活有关。This study aims to investigate the mechanism of Mahuang Lianqiao Chixiaodou Decoction(MHLQ)and its main active components in treating immunoglobin A nephropathy(IgAN).The rat model of IgAN was established by a combination of measures including gavage of bovine serum albumin,subcutaneous injection of carbon tetrachloride,and tail vein injection of lipopolysaccharide.The modeled rats were randomized into model,low-,medium-,and high-dose(1.773,3.545,and 7.090 g·kg^(-1),respectively)MHLQ,phillyrin(PHI,0.020 g·kg^(-1)),pseudoephedrine(PSE,0.020 g·kg^(-1)),and losartan potassium(LP,9.003 mg·kg^(-1))groups,and Wistar rats were used as the control.Rats were administrated with corresponding drugs by gavage,and those in the control and model groups received an equal volume of normal saline.All the groups were treated for 4 consecutive weeks.Urine,serum,liver,and kidney samples were collected from rats in each group at the end of drug administration.The 24 h urine protein and renal function were examined,and staining was performed to observe the pathological changes in the renal tissue.The immunofluorescence assay was employed to detect the expression of IgA and complement C3/C3b/C3c in the renal tissue.Electron microscopy was employed to observe the ultrastructure of the renal tissue.Enzyme-linked immunosorbent assay was performed to determine the expression of complement C3 and sublytic C5b-9 in the serum and renal tissue.Western blot was performed to determine the expression levels of hepatic and renal complement C3/C3b/C3c,C5/C5a,C5b-9,and complement factor B(CFB).Immunohistochemistry(IHC)was employed to measure the expression of complement C3 in the renal tissue.The results showed that compared with the control group,the model group had elevated levels of blood urea nitrogen and serum creatinine,proliferation of glomerular mesangial cells and extracellular matrix,and glomerular deposition of IgA immune complexes or electron-dense material.In addition,the model group showcased increased serum C3 levels and up-regula

关 键 词：麻黄连翘赤小豆汤 IGA肾病 补体激活 炎症反应 补体C3 

分 类 号：R285.5[医药卫生—中药学]