摘要
为了探究三七皂苷R_(1)(notoginsenoside R_(1),NGR_(1))干预肾脏氧化应激和Nrf2/HO-1信号通路改善IgA肾病(immunoglobulin A nephropathy,IgAN)模型小鼠肾损伤的作用和机制,通过连续牛血清白蛋白(bovine serum albumin,BSA)灌胃、皮下注射四氯化碳(carbon tetrachloride,CCl4)蓖麻油、尾静脉注射脂多糖(lipopolysaccharide,LPS)等综合措施建立IgAN小鼠模型,成模后,将IgAN模型鼠随机分为模型组,NGR_(1)低、中、高剂量组以及氯沙坦(losartan)组,并以C57BL6小鼠为正常对照。模型组、正常组使用磷酸缓冲盐溶液(phosphate buffered saline,PBS)灌胃,NGR_(1)各组使用不同剂量NGR_(1)灌胃,氯沙坦组使用losartan灌胃,连续干预4周。末次给药后收集各组小鼠24 h尿液,动物实验结束时采集小鼠血清和肾组织标本。检测小鼠尿红细胞计数(urine red blood cell count,URBCC)、24 h尿蛋白、血清肌酐(serum creatinine,Scr)、尿素氮(blood urea nitrogen,BUN)的水平,ELISA检测小鼠血清中半乳糖缺陷的IgA1(galactose-deficient IgA1,Gd-IgA1)、肾损伤分子1(kidney injury molecule 1,Kim-1)、中性粒细胞明胶酶相关脂质运载蛋白(neutropil gelatinase-associated lipocalin,NGAL)水平,试剂盒检测丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)的水平,免疫荧光(immunofluorescence,IF)检测肾小球系膜区谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)的表达,Western blot检测肾组织中核转录因子E2相关因子2(nuclear transcription factor E2 related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase 1,HO-1)信号通路蛋白表达,苏木精-伊红(hematoxylin-eosin staining,HE)染色法观察小鼠肾小球区域病理改变。结果显示,与模型组比较,NGR_(1)高剂量组血清Gd-IgA1水平、URBCC、24 h尿蛋白水平、肾损伤指标(Kim-1、NGAL)明显降低,肾功能指标(BUN、Scr)明显改善;肾脏组织中SOD活性、GPX4表达升高,而MDA下降,Nrf2和HO-1蛋白表达升高;同时,肾组织H
The purpose of this study was to investigate the effect of notoginsenoside R_(1)(NGR_(1))on alleviating kidney injury by regulating renal oxidative stress and the Nrf2/HO-1 signaling pathway in mice with IgA nephropathy(IgAN)and its mechanism.The mouse model of IgAN was established using a variety of techniques,including continuous bovine serum albumin(BSA)gavage,subcutaneous injections of carbon tetrachloride(CCl4)castor oil,and tail vein injections of lipopolysaccharide(LPS).After successful modeling,mice with IgAN were randomly separated into a model group,low,medium,and high-dose NGR_(1) groups,and a losartan group,and C57BL6 mice were utilized as normal controls.The model and normal groups were given phosphate buffered saline(PBS)by gavage,the NGR_(1) groups were given varying dosages of NGR_(1) by gavage,and the losartan group was given losartan by gavage for 4 weeks.The 24-hour urine of mice was collected after the last administration,and serum and kidney tissues of mice were taken at the end of the animal experiment.Then urine red blood cell count(URBCC),24-hour urine protein(24 h protein),serum creatinine(Scr),and blood urea nitrogen(BUN)levels were measured.The enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of galactose-deficient IgA1(Gd-IgA1),kidney injury molecule 1(Kim-1),and neutropil gelatinase-associated lipocalin(NGAL)in the mouse serum.The assay kits were used to detect the levels of malondialdehyde(MDA)and superoxide dismutase(SOD),and immunofluorescence(IF)was used to detect the expression level of glutathione peroxidase 4(GPX4)in the mesangial region.Western blot was used to detect the protein expression of nuclear transcription factor E2 related factor 2(Nrf2)/heme oxygenase 1(HO-1)signaling pathway in the renal tissue.Hematoxylin-eosin(HE)staining was used to observe pathological alterations in the glomerulus of mice.The results revealed that,as compared with the model group,the serum Gd-IgA1 level,URBCC,24 h protein level,renal damage markers(Kim-1 and NGAL)in the hi
作者
吴晶魁
陈冬平
叶朝阳
WU Jing-kui;CHEN Dong-ping;YE Chao-yang(Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201200,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第14期3839-3847,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81873617,82170747)。
