摘要
目的:基于转录组测序(RNA-seq)技术分析益肾散结复方抑制阿霉素肾病肾小球硬化的作用机制。方法:将40只SD大鼠随机分为4组,分别为正常组、阿霉素模型组、盐酸贝那普利对照组和益肾散结复方组(以下统称正常组、模型组、对照组和复方组),连续2次尾静脉注射盐酸阿霉素制备阿霉素大鼠肾病模型,对照组和复方组分别按10mg/kg/d盐酸贝那普利和6.24g/kg/d益肾散结复方煎剂灌胃6周,其余组灌胃予等剂量生理盐水,灌胃前后对比大鼠一般情况、24小时尿蛋白定量(24h UTP)、血尿素氮(BUN)及血肌酐(SCr)指标变化,电镜下观察大鼠肾脏病理改变;同时将复苏的HBZY-1大鼠肾小球系膜细胞分为以上4组,以40μM浓度阿霉素诱导大鼠肾脏系膜细胞凋亡模型,分别对应处理各组细胞,观察大鼠肾小球系膜细胞凋亡情况。在此基础上,对正常组、模型组、复方组细胞进行RNA测序,筛选差异表达基因(DEG),对共同DEG进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)富集分析。结果:动物实验研究显示益肾散结复方可改善阿霉素肾病大鼠便溏、反应迟钝、活动减少等精神状态,减少24h UTP、BUN和SCr水平( P <0.05),减少阿霉素肾病大鼠肾小球系膜区基质沉积,改善基底膜缺血、皱缩情况,从而抑制肾小球硬化程度;细胞实验研究显示:益肾散结复方可延缓阿霉素诱导的HBZY-1大鼠肾小球系膜细胞凋亡过程,从而对肾小球硬化起抑制作用。根据细胞RNA-seq结果,益肾散结复方抗阿霉素肾病肾小球硬化的共同DEG有863个,542个上调, 321个下调;GO分析显示这些DEG主要富集在生物过程、细胞成分和分子功能三个方面,其中与肾小球硬化相关的GO功能有:正向调节细胞迁移、细胞粘附、细胞外基质、整合素结合、CXCR趋化因子受体结合、趋化因子活动等;KEGG分析显示上述DEG富集在19条通路( P <0.05)中,其中细胞外基质(ECM)
Objective: To analyze the mechanism of Yishensanjie compound in inhibiting glomerulosclerosis of adriamycin nephropathy based on transcriptome sequencing(RNA-seq). Method: Forty SD rats were randomly divided into 4groups, which were normal group, adriamycin model group, benazepril hydrochloride control group and Yishensanjie compound group(hereinafter referred to as normal group, model group, control group and compound group), and adriamycin hydrochloride was injected into the tail vein twice to prepare adriamycin rat nephropathy model. The control group and compound group were given 10mg/kg/d benazepril hydrochloride and 6.24g/kg/d Yishensanjie compound decoction for 6weeks, and the other groups were given the same dose of normal saline. The general situation, 24h urinary protein quantity(24h UTP), blood urea nitrogen(BUN) and blood creatinine(SCr) indexes of the rats were compared before and after gavage. The pathological changes of kidney were observed under electron microscope. At the same time, the revived HBZY-1rat mesangial cells were divided into the above 4groups, and the apoptosis model of rat renal mesangial cells was induced by doxorubicin at 40μM concentration, and the cells of each group were treated correspondingly, and the apoptosis of rat mesangial cells was observed. On this basis, the normal group, model group and compound group cells were sequenced by RNA, screened for differentially expressed genes(DEG), and the common DEG was enriched by gene ontology(GO) and kyoto encyclopedia of genes and genomes(KEGG). Results: The animal experimental study showed that Yishenanjie compound could improve the mental states of lax stoses, dull reaction and decreased activity in rats with doxorubicin nephropathy, reduce the levels of UTP, BUN and SCr for 24h( P <0.05), reduce the matrix deposition in mesangial region of the glomerulus in rats with doxorubicin nephropathy, improve the condition of basement membrane ischemia and crease, and inhibit the degree of glomerular sclerosis. The cell experiment showed
作者
曹慧慧
屈凯
王悦彤
柳小远
赵亚峰
于小勇
范钰
毛加荣
CAO Huihui;QU Kai;WANG Yuetong(Xi an Chang an District Hospital of Traditional Chinese Medicine,Xi an Shaanxi 710100,China;Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046,China;Shaanxi Provincial Hospital of Traditional Chinese Medicine,Xi an Shaanxi 710003,China;Shaanxi Institute of Traditional Chinese Medicine,Xi an Shaanxi 710003,China)
出处
《四川中医》
2024年第11期60-67,共8页
Journal of Sichuan of Traditional Chinese Medicine
基金
国家自然科学基金项目(编号:81603454)。
关键词
转录组测序技术
益肾散结复方
肾小球硬化
阿霉素肾病
细胞外基质-受体相互作用
Transcriptome sequencing technology
Yishen sanjie compound
Glomerulosclerosis
Adriamycin nephropathy
Extracellular matrix-receptor interaction
