探讨固本通络方基于细菌脂多糖介导的肠B细胞激活对Gd-IgA1的影响  

Exploration of the Effect of Guben Tongluo Decoction(固本通络方)on the Activation of Intestinal B Cells Mediated by Bacterial LPS on Gd-IgA1

作　　者：汪容 沈沛成[2] 何立群[2] 吴卿[2] WANG Rong;SHEN Peicheng;HE Liqun;WU Qing(The First Affiliated Hospital of Hunan College of Traditional Chinese Medicine,Zhuzhou 412000,Hunan,China;Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]湖南中医药高等专科学校附属第一医院,湖南株洲412000 [2]上海中医药大学附属曙光医院,上海201203

出　　处：《辽宁中医药大学学报》2024年第2期47-53,F0003,共8页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(81904124);上海市浦东新区卫生系统重点学科群建设项目(PWZxq2017-07);上海市进一步加快中医药事业发展三年行动计划项目[ZY(2018-2020)-CCCX-2003-08,ZY(2018-2020)-FWTX-7005]。

摘　　要：目的探讨固本通络方对IgA肾病小鼠肠道细菌脂多糖(LPS)和糖基化缺陷IgA1(Gd-IgA1)的干预及肾保护作用。方法将36只雄性小鼠分为正常对照组、模型组、固本通络方低剂量组、固本通络方中剂量组、固本通络方高剂量组、依立托仑组。实验采用ddY小鼠构建IgA肾病动物模型,将模型组和各药物组进行造模,中药干预各组分别予以固本通络方免煎颗粒低(13.2 g/kg)、中(26.4 g/kg)、高(52.8 g/kg)3种不同剂量药液灌胃给药,正常对照组和模型组小鼠给予0.9%氯化钠溶液灌胃(10 mL/kg),依立托仑组予以腹腔注射依立托仑药液,各药物组每天给予药物1次,连续8周。干预4周、8周后,测定各组24 h尿蛋白定量;8周测定血清及肠道黏膜LPS、IgA、GdIgA1水平,观察肾脏病理染色及右肾IgA沉积情况,实时荧光定量PCR检测Cosmc基因表达,Western blot法测定Cosmc蛋白表达及IgA肠道菌群特异性,流式细胞术检测肠道B淋巴细胞活化率及分泌IgA前体B淋巴细胞比率,对肠道黏膜组织LPS水平与B细胞活化率及IgA前体B细胞比率行相关性分析。结果固本通络方可降低24 h尿蛋白定量水平及血清肌酐水平(P<0.05)。HE染色发现,与模型组相比,固本通络方用药组肾小球系膜细胞、内皮细胞及基质增生显著减少,肾小球系膜区IgA沉积显著改善;血清及肠道黏膜LPS水平检测发现,与模型组比较,各中药干预组均能降低小鼠血清和肠道黏膜组织LPS、Gd-IgA1和IgA水平(P<0.05);降低B细胞活化率及IgA前体B细胞比率(P<0.05),qRT-PCR和Western blot结果显示固本通络方能够显著降低小鼠粪便培养中菌落蛋白表达水平,上调Cosmc蛋白和Cosmc基因表达水平(P<0.05)。LPS水平与B细胞活化率及IgA前体B细胞比率均呈正相关(P<0.05)。结论固本通络方能有效降低IgA肾病小鼠蛋白尿及血清肌酐水平,减轻肾脏病理损伤及肾小球系膜区IgA沉积,其作用可能和固本通络方降低IgA�Objective To investigate the intervention and renal protection of Guben Tongluo Decoction(固本通络方)on intestinal bacteria LPS and galactose-deficient IgA1(Gd-IgA1)in mice with IgA nephropathy.Methods In this experiment,ddY mice were used to construct the animal model of IgA nephropathy,according to the random number table,the mice were divided into normal control group,model group,Guben Tongluo Decoction low dose group,Guben Tongluo Decoction medium dose group,Guben Tongluo Decoction high dose group and Elitoren group.The intervention groups of traditional Chinese medicine were given Guben Tongluo Decoction granules by gavage at three different doses(13.2,26.4,52.8 g/kg).Mice in normal control group and model group were given 0.9%sodium chloride solution(10 mL/kg),and Elitoren group was given intraperitoneal injection of Elitoren,once a day for 8 weeks.At the 4th and 8th weeks,24-hour urinary protein was measured.At the 8th week,the levels of LPS,IgA and Gd-IgA1 in serum and intestinal mucosa were measured,the pathological changes of kidney and IgA deposition in right kidney were observed,the expression of Cosmc gene was detected by fluorescence quantitative PCR,the expression of Cosmc protein and the specificity of IgA intestinal flora were measured by Western blotting,and the activated proportion of intestinal B lymphocytes and the proportion of B lymphocytes secreting IgA precursor were detected by cell flow cytometry.Results Compared with the model group,each traditional Chinese medicine intervention group could reduce the levels of LPS,Gd-IgA1 and IgA in serum and intestinal mucosa(P<0.05);It decreased the activation rate of B cells and the proportion of IgA precursor B cells(P<0.05),decreased the expression level of colony protein in mouse fecal culture,increased the expression level of Cosmc protein and Cosmc mRNA(P<0.05),and decreased the 24-hour protein quantity and serum creatinine level(P<0.05).Pathologically,the proliferation of glomerular mesangial cells,endothelial cells and matrix in each Gu

关 键 词：固本通络方 IGA肾病 糖基化缺陷IgA1 肠道细菌脂多糖 

分 类 号：R256.5[医药卫生—中医内科学]