商陆致大鼠肾损伤的基因组学研究  

Genomics Research on Phytolaccae Radix Induced Renal Injury in Rats

作　　者：徐婷婷[1,2] 姚广涛 金若敏[1] 唐黎明[3] 李一飞[1] 羊菲[1] XU Tingting;YAO Guangtao;JIN Ruomin;TANG Liming;LI Yifei;YANG Fei(Center for Drug Safety Evaluation and Research,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai InnoStar Bio-Tech Co.,LTD,Shanghai 201203,China;Department of Pharmacology&Toxicology,Shanghai Institute for Food and Drug Control,Shanghai 201203,China)

机构地区：[1]上海中医药大学药物安全评价研究中心,上海201203 [2]上海益诺思生物技术股份有限公司,上海201203 [3]上海市食品药品检验所药理毒理室,上海201203

出　　处：《中药新药与临床药理》2021年第11期1591-1600,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家重点基础研究发展计划项目(973计划,2009CB522807);国家重点研发计划项目(SQ2017YEC170204):基于器官芯片技术的中药安全性有效性评价体系。

摘　　要：目的采用寡核苷酸微阵列芯片技术探讨商陆长期给药对大鼠肾组织相关细胞因子及基因表达的影响,以阐释商陆致大鼠肾损伤的作用机制。方法将大鼠随机分为对照组及商陆低、高剂量组(10、20 g·kg-1),灌胃商陆水煎液(20 mL·kg-1)或等体积纯水,每天1次,连续49 d。采用酶联免疫吸附法(ELISA)和免疫组化法(IHC)检测肾组织中相关炎性因子[白细胞介素(IL)1β、IL-6、肿瘤坏死因子α(TNF-α)]含量及核因子κB(NF-κB)表达;采用全基因组寡核苷酸生物芯片检测大鼠肾组织基因表达,筛选差异基因,并进行京都基因与基因组百科全书(KEGG)通路分析;采用逆转录-聚合酶链反应(RT-PCR)法验证差异基因。结果与对照组比较,商陆高、低剂量组大鼠肾组织中IL-1β、IL-6、TNF-α含量均明显升高(P<0.05,P<0.01),商陆高剂量组大鼠肾组织中NF-κB表达量明显升高(P<0.05);商陆高、低剂量组分别有2 560、1 923个差异表达基因,两组均激活了丝裂原活化蛋白激酶(MAPK)信号通路中的MAP、p38 MAPK、JNK 3条信号通路,且高剂量组引起MAPK通路的差异基因数是低剂量组的2倍;商陆高剂量组还激活了Janus激酶/信号转导与转录激活因子(JAK-STAT)信号通路及细胞凋亡(Apoptosis)信号通路。验证实验结果表明,与对照组比较,商陆低剂量组大鼠肾组织的转化生长因子β1(TGF-β1)基因表达量明显升高(P<0.01),而高剂量组大鼠肾组织的IL-1β、表皮生长因子受体(EGFR)、TGF-β1基因表达量均显著升高(P<0.01)。结论通过IL-1β、TGF-β1/β2等细胞因子激活MAPK、JAK-STAT及细胞凋亡信号转导通路,继而引起炎性反应和细胞凋亡,可能是商陆致大鼠肾损伤的机制之一。Objective To study the effects of long-term administration of Phytolaccae radix on cytokine content and gene expression in rat’s renal tissue based on oligonucleotide microarray chips,and discuss the mechanism of Phytolaccae radix induced renal injury in rats.Methods Rats were randomly divided into control group,Phytolaccae radix groups of low-and high-dose,which were treated with pure water or Phytolaccae radix decoction at doses of 10 g·kg-1 and 20 g·kg-1 by intragastric administration respectively for 49 consecutive days.The inflammatory factors,Interleukin-1β(IL-1β),IL-6,and Tumor Necrosis Factor-α(TNF-α),and the expression of Nuclear Factor kappa-B(NF-κB) in renal tissue were detected by Enzyme-Linked Immunosorbent Assay(ELISA) and immunohistochemistry(IHC).The gene expression in rat kidney was detected by oligonucleotide microarray chip technology,the differential genes were screened and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis was performed,and the differential genes were verified by Reverse Transcription-Polymerase Chain Reaction(RT-PCR).Results Compared with that in the control group,the protein contents of inflammatory mediators including IL-1β,IL-6,and TNF-α in the Phytolaccae radix groups with low-and high-dose were increased significantly(P<0.05,P<0.01),and the expression of NF-κB in the high-dose group was increased significantly(P<0.05).A total of 2 560 and 1 923 differential gene expression were found in the Phytolaccae radix groups with high-and low-dose respectively,signal pathways of MAP,JNK,and p38 in Mitogen-activated Protein Kinase(MAPK)signaling pathway were all activated in high-and low-dose groups,the number of differential genes in high dose group was twice as many as that in low dose group.Additionally,Janus Kinase/Signal Transducer and Activator of Transcription(JAK/STAT)and apoptosis signal pathways were also activated in the high-dose group.Compared with that in the control group,the expression levels of IL-1β,EGFR,and TGF-β1 genes in the highdo

关 键 词：商陆 肾损伤 炎性因子 基因组学 炎性反应 细胞凋亡 大鼠 

分 类 号：R285.5[医药卫生—中药学] R994.1[医药卫生—中医学]