基于C1GALT1/C1GALT1C1通路探讨加味升降散对IgA肾病大鼠Gd-IgA1的影响  

Effect of Modified Shengjiangsan on Gd-IgA1 in IgA Nephropathy Rats:Based on C1GALT1/C1GALT1C1 Signaling Pathway

作　　者：张圆圆 靳培培 靳贺超 顾悦 郭登洲 ZHANG Yuanyuan;JIN Peipei;JIN Hechao;GU Yue;GUO Dengzhou(Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;The First Affiliated Hospital of Hebei University of Chinese Medicine,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China)

机构地区：[1]河北中医学院研究生学院,石家庄050200 [2]河北中医学院第一附属医院,河北省中医院,石家庄050011

出　　处：《中国实验方剂学杂志》2022年第14期70-80,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省科技支撑计划项目(16277765D);河北中医学院科技能力提升项目(KTZ2019025)。

摘　　要：目的:通过观察加味升降散对免疫球蛋白(Ig)A肾病大鼠血清及肾组织中微小RNA-148b(miRNA-148b)、白细胞介素-6(IL-6)、核心1β1,3-半乳糖基转移酶(C1GALT1)、分子伴侣Cosmc(core1β3-Gal-T特异性分子伴侣,C1GALT1C1)、半乳糖缺乏IgA1(Gd-IgA1)表达的影响探讨其可能作用机制,为临床运用加味升降散治疗IgA肾病提供科学依据。方法:将42只SPF级雄性SD大鼠按照随机数字表法分为正常组8只、造模组34只,将大鼠适应性饲养1周后采用联合牛血清白蛋白(BSA)灌胃+尾静脉注射脂多糖(LPS)+皮下注射四氯化碳(CCl4)、蓖麻油的免疫复合的方法建立实验性IgA肾病大鼠模型。造模完成后随机选取2只大鼠取材,检测造模是否成功。造模成功后,将造模组大鼠按照随机数字表法分成模型组、中药低剂量(加味升降散,6.27 g·kg;)组、中药高剂量(加味升降散,12.54 g·kg;)组、贝那普利(10 mg·kg^(-1))组各8只,进行药物灌胃,1次/d,治疗4周。于实验第1周末、第9周末、第13周末检测24 h尿蛋白定量(24 h-UTP),第14周各组大鼠麻醉后取材,股动脉取血后离心取上清检测白蛋白(ALB)、天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、血肌酐(SCr)、尿素氮(BUN);酶联免疫吸附测定法(ELISA)检IL-6、Gd-IgA1表达水平变化;苏木素-伊红(HE)/马松(Masson)/碘酸六胺银(PASM)法观察肾组织病理改变;透射电镜观察肾小球超微结构变化;免疫组化法观察IL-6、C1GALT1、C1GALT1C1表达情况;免疫荧光观察肾小球系膜区;实时荧光定量聚合酶链式反应(Real-time PCR)观察miRNA-148b、IL-6、C1GALT1、C1GALT1C1 mRNA表达;蛋白免疫印迹法(Western blot)检测IL-6、C1GALT1、C1GALT1C1蛋白水平变化。结果:与正常组比较,模型组大鼠24 hUTP、SCr、ALT、IL-6、Gd-IgA1含量水平明显升高(P<0.05),ALB含量水平明显降低(P<0.05);肾小球系膜细胞增生、系膜区增厚、足细胞足突融合,大量颗粒状IgA免疫复合物�Objective:The effect of modified Shengjiangsan on immunoglobulin A(IgA)nephropathy was observed.The microRNA-148b(miRNA-148b),interleukin 6(IL-6),core 1 beta 1,3-galactosyltransferase(C1GALT1),molecular chaperone Cosmc(core1β3-Gal-T-specific molecular chaperone C1GALT1C1),and galactose-deficient IgA1(Gd-IGA1)in serum and kidney tissues of IgA nephropathy rats were detected to explore the underlying mechanism.The result is expected to lay a scientific basis for clinical application of modified Shengjiangsan in the treatment of IgA nephropathy.Method:A total of 42 SPF male SD rats were randomized into the normal group(8rats)and modeling group(34 rats)with the random number table method.After one week of adaptive feeding,rats for modeling were given bovine serum albumin(BSA,gavage),lipopolysaccharide(LPS,injection into tail vein),carbon tetrachloride(CCl4,subcutaneous injection),and castor oil to induce IgA nephropathy.After modeling,two rats were randomly selected to test the modeling outcome.Then the model rats were classified into the model group,low-dose Chinese medicine group(modified Shengjiangsan,6.27 g·kg^(-1)),high-dose Chinese medicine group(modified Shengjiangsan,12.54 g·kg^(-1)),and benazepril group(10 mg·kg^(-1))with the random number table method,8 in each group.The administration(gavage,once a day)lasted 4 weeks.The 24-h urinary total protein(24 h-UTP)was detected at the end of the 1st,9th,and 13th week of the experiment.At the 14th week,after anesthesia,femoral artery blood was collected and centrifugated.The supernatant was collected to detect albumin(ALB),aspartate aminotransferase(AST),alanine aminotransferase(ALT),serum creatinine(SCr),and blood urea nitrogen(BUN).The expression levels of IL-6 and Gd-IGA1 were determined by enzyme-linked immunosorbent assay(ELISA).Based on hematoxylin-eosin(HE)/Masson/periodic Schiff-methenamine silver(PASM)staining,the pathological changes of renal tissues were observed.Ultrastructural changes of glomeruli were observed by transmission electron microscopy.Th

关 键 词：免疫球蛋白(Ig)A肾病 加味升降散 微小RNA-148b 白细胞介素-6 核心1β1 3-半乳糖基转移酶 核心1β3-Gal-T特异性分子伴侣 半乳糖缺乏IgA1 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R256.5R692