凉血解毒活血方治疗大鼠肾毒血清肾炎新月体病变  

Liangxue Jiedu Huoxue Recipe for Crescentic Lesions in Rats with Nephrotoxic Serum Nephritis

作　　者：张冰[1] 黄岩杰[1,2] 侯改灵 彭超群 刘萌萌 ZHANG Bing;HUANG Yanjie;HOU Gailing;PENG Chaoqun;LIU Mengmeng(School of Pediatrics,Henan University of Chinese Medicine,Zhengzhou Henan China 450046;The First Affiliated Hospital to Henan University of Chinese Medicine,Zhengzhou Henan China 450000;Children′s Hospital Affiliated to Zhengzhou University/Children′s Hospital of Henan Province,Zhengzhou Henan China 450018)

机构地区：[1]河南中医药大学儿科医学院,河南郑州450046 [2]河南中医药大学第一附属医院,河南郑州450000 [3]郑州大学附属儿童医院/河南省儿童医院,河南郑州450018

出　　处：《中医学报》2024年第1期181-188,共8页Acta Chinese Medicine

基　　金：国家自然科学基金项目(82174187);河南省中医药学科领军人才项目[豫卫中医函(2021)8号];2022年中原科技创新领军人才项目(234200510028)。

摘　　要：目的:探讨凉血解毒活血方治疗大鼠肾毒血清肾炎(nephrotoxic serum nephritis,NTSN)新月体病变的疗效特点。方法:45只SPF级雄性Wistar-Kyoto(WKY)大鼠随机分为正常组、模型Ⅰ组(造模第7天取材)、模型Ⅱ组、甲泼尼龙组和凉血解毒活血方+甲泼尼龙组,每组9只。建立大鼠NTSN模型的方法为:从Sprague Dawley大鼠中提取肾小球基底膜(glomerular basement membrane,GBM)抗原,在新西兰白兔背部行对称多点皮下注射,制备肾毒血清。WKY大鼠预免疫7 d后,尾静脉注射肾毒血清,连续注射3 d即可。除模型Ⅰ组外,其余各组大鼠均在造模第7天开始灌胃,持续7 d,并在末次灌胃后留取24 h尿液。应用Periodic Acid-Schiff(PAS)染色观察各组大鼠肾脏组织病理变化,包括新月体百分比。使用全自动生化分析仪检测各组大鼠24小时尿蛋白定量、尿免疫球蛋白G(immunoglobulin G,IgG)、尿肌酐(creatinine,Cr)水平。采用免疫组织化学法检测各组大鼠肾脏组织整合素β2(integrin beta-2,ITGB2)蛋白定位及表达水平。结果:正常组大鼠肾小球及周围小管细胞排列整齐,形态正常,边界清晰,未见明显损伤;模型Ⅰ组大鼠在造模第7天时,25%~50%的肾小球形成细胞性新月体,新月体内可见纤维蛋白沉积;模型Ⅱ组大鼠在造模第14天时,肾小球内出现大量细胞性新月体病变,新月体比例≥50%。与正常组比较,模型Ⅰ组大鼠的24小时尿蛋白定量、新月体百分比均显著增加(P<0.05);与模型Ⅰ组比较,模型Ⅱ组大鼠的24小时尿蛋白定量、新月体百分比均显著增加(P<0.05)。与正常组比较,模型Ⅱ组大鼠的24小时尿蛋白定量、新月体百分比与尿IgG/Cr比值均明显升高(P<0.05);与模型Ⅱ组比较,甲泼尼龙组和凉血解毒活血方+甲泼尼龙组大鼠的24小时尿蛋白定量、新月体百分比与尿IgG/Cr比值均明显降低(P<0.05);与甲泼尼龙组比较,凉血解毒活血方+甲泼尼龙组大鼠的24小时尿蛋白定Objective:To characterize the efficacy of Liangxue Jiedu Huoxue Fang(LJH)in treating crescentic lesions in rats with nephrotoxic serum nephritis(NTSN).Method:45 Wistar Kyoto rats were randomly divided into a normal group,ModelⅠgroup(taken on the 7th day of modeling),ModelⅡgroup,methylprednisolone group,and LJH+methylprednisolone group,with 9 rats in each group.The method for establishing a rat NTSN model is to extract the glomerular basement membrane(GBM)antigen from Sprague Dawley rats and perform symmetrical multi-point subcutaneous injection on the back of New Zealand white rabbits to prepare nephrotoxic serum.After 7 days of pre immunization in WKY rats,renal toxic serum was injected into the tail vein continuously for 3 days.Except for ModelⅠgroup,all other groups started gastric lavage on the 7th day of modeling,lasting for 7 days,and urine was collected for 24 hours after the last gastric lavage.Apply PeriodicAcid Schiff(PAS)staining to observe the pathological changes in renal tissue of rats in each group,including the percentage of crescents.Use a fully automated biochemical analyzer to detect the 24-hour urine protein content,urine immunoglobulin G(IgG)and creatinine(Cr)levels in each group of rats.Detection of integrin in renal tissue of rats in each group using immunohistochemical methodβLocalization and expression level of integrinbeta-2(ITGB2)protein.Results:The glomeruli and surrounding tubular cells of the normal group rats were arranged neatly,with normal morphology and clear boundaries,and no significant damage was observed;On the 7th day of modeling in Model GroupⅠrats,25%to 50%of the glomeruli formed cellular crescents,and fibrin deposition was visible in the crescents;On the 14th day of modeling,a large number of cellular crescentic lesions appeared in the glomeruli of ModelⅡrats,with a crescentic body proportion of≥50%.Compared with the normal group,the 24-hour urine protein quantification and crescent percentage in ModelⅠgroup were significantly increased(P<0.05);Compared wit

关 键 词：凉血解毒活血方 甲泼尼龙 肾毒血清肾炎 新月体肾小球肾炎 ITGB2 IGG 

分 类 号：R285.5[医药卫生—中药学]