药学学报, 2019, 54(8): 1439-1448
引用本文:
贺升升, 李爱平, 张王宁, 秦雪梅. 基于转录组测序技术探讨黄芪水提物干预多柔比星肾病大鼠的分子机制[J]. 药学学报, 2019, 54(8): 1439-1448.
HE Sheng-sheng, LI Ai-ping, ZHANG Wang-ning, QIN Xue-mei. Exploring the molecular mechanism of aqueous extract from Huang qi on doxorubicin induced nephropathy in rats using transcriptome sequencing technique[J]. Acta Pharmaceutica Sinica, 2019, 54(8): 1439-1448.

基于转录组测序技术探讨黄芪水提物干预多柔比星肾病大鼠的分子机制
贺升升1,2, 李爱平1, 张王宁1, 秦雪梅1
1. 山西大学中医药现代研究中心, 山西 太原 030006;
2. 山西大学化学化工学院, 山西 太原 030006
摘要:
为了研究黄芪水提物对多柔比星肾病大鼠基因表达谱的影响,探讨黄芪水提物干预多柔比星肾病大鼠的分子机制。采用转录组测序技术检测对照组、模型组和黄芪水提物组大鼠肾组织的基因表达谱,通过STEM趋势分析软件筛选趋势表达的差异表达基因(differentially expressed genes,DEGs),并针对DEGs进行GO功能富集与KEGG通路分析,用实时荧光定量PCR(RT-qPCR)对基因的表达量进行验证。实验过程中对动物的处置符合动物实验伦理标准。结果显示,与对照组相比,模型组共筛选DEGs 432个;与模型组相比,黄芪水提物组共筛选DEGs811个。KEGG通路分析与荧光定量PCR结果指示,PI3K-AKT通路(Col6a6、Nr4a1、Sgk1、Gng7)与脂质代谢相关基因(Cpt1b、Pcsk9、Abca1、Ascm5)是黄芪水提物治疗多柔比星肾病、发挥肾脏保护作用的关键通路与基因。总之,黄芪水提物干预多柔比星肾病大鼠的分子机制与凋亡相关基因和脂质代谢相关基因密切相关,此结果为后续黄芪治疗多柔比星肾病的关键基因验证和作用机制研究提供了研究基础。
关键词:    黄芪      水提物      多柔比星肾病      转录组学      分子机制     
Exploring the molecular mechanism of aqueous extract from Huang qi on doxorubicin induced nephropathy in rats using transcriptome sequencing technique
HE Sheng-sheng1,2, LI Ai-ping1, ZHANG Wang-ning1, QIN Xue-mei1
1. Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan 030006, China;
2. College of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006, China
Abstract:
We studied the effect of aqueous extract from Huang qi on gene expression profile of doxorubicin induced nephropathy in rats, and explored the molecular mechanism of the intervention. The gene expression profiles of control group, model group and aqueous extract from Huang qi group were detected by using transcriptome sequencing technique. The differentially expressed genes (DEGs) were screened by STEM trend analysis software. GO function enrichment and KEGG pathway analysis were performed for DEGs, and the gene expression level was verified by real-time fluorescence quantitative PCR (RT-qPCR). The results showed that, compared with the control group, 432 DEGs were obtained in doxorubicin nephropathy model group; compared with the model group, 811 DEGs were obtained due to aqueous extract of Huang qi. The results of GO function enrichment and KEGG enrichment analysis indicated that PI3K-AKT pathway (Col6a6, Nr4a1, Sgk1, Gng7) and lipid metabolism-related genes (Cpt1b, Pcsk9, Abca1, Ascm5) were the key pathways and genes in the treatment of doxorubicin induced nephropathy by aqueous extract from Huang qi, which played a protective role in kidney. In conclusion, the molecular mechanism of aqueous extract from Huang qi in protection against doxorubicin induced nephropathy rats is closely related to apoptosis-related genes and lipid metabolism-related genes, suggesting for the need of follow-up study for key gene validation and mechanism of action of aqueous extract from Huang qi for prevention of doxorubicin induced nephropathy.
Key words:    Huang qi    aqueous extract    doxorubicin nephropathy    transcriptomics    molecular mechanism   
收稿日期: 2019-04-01
DOI: 10.16438/j.0513-4870.2019-0225
基金项目: 国家自然科学基金资助项目(31570346).
通讯作者: 李爱平,Tel:86-351-7011501,E-mail:qinxm@sxu.edu.cn;秦雪梅,Tel:86-351-7018379,E-mail:aipingli@sxu.edu.cn
Email: qinxm@sxu.edu.cn;aipingli@sxu.edu.cn
相关功能
PDF(846KB) Free
打印本文
0
作者相关文章
贺升升  在本刊中的所有文章
李爱平  在本刊中的所有文章
张王宁  在本刊中的所有文章
秦雪梅  在本刊中的所有文章

参考文献:
[1] He SS, Li AP, Zhang WN, et al. Research progress on adriamycin nephropathy model and its pathological mechanism[J]. Chin Tradit Herb Drugs (中草药), 2018, 49:5426-5434.
[2] Hulkko J, Patrakka J, Lal M, et al. Neph1 is reduced in primary focal segmental glomerulosclerosis, minimal change nephrotic syndrome, and corresponding experimental animal models of adriamycin-induced nephropathy and puromycin aminonucleoside nephrosis[J]. Nephron Extra, 2014, 4:146-154.
[3] Kawamorita Y, Shiraishi T, Tamura Y, et al. Renoprotective effect of topiroxostat via antioxidant activity in puromycin aminonucleoside nephrosis rats[J]. Physiol Rep, 2017, 5:e13358.
[4] Jiang Q, He X, Zou Y, et al. Altered gut microbiome promotes proteinuria in mice induced by adriamycin[J]. AMB Expr, 2018, 8:31.
[5] Vaziri ND. Disorders of lipid metabolism in nephrotic syndrome:mechanisms and consequences[J]. Kidney Int, 2016, 90:41-52.
[6] Zhong F, Wang W, Lee K, et al. Role of C/EBP-α in adriamycininduced podocyte injury[J]. Sci Rep, 2016, 6:33520.
[7] Jia L, Ma X, Gui B, et al. Sorafenib ameliorates renal fibrosis through inhibition of TGF-β-induced epithelial-mesenchymal transition[J]. PLoS One, 2015, 10:e0117757.
[8] Qian ZZ, Dan Y, Liu YZ, et al. Pharmacopoeia of the People's Republic of China (2010 Edition):a milestone in development of China's healthcare[J]. Chin Herbal Med, 2010, 2:157-160.
[9] Yang L, Li AP, Zhang WN, et al. Research progress on pharmacological effects and clinical application of single Astragali Radix and classical formulae containing Astragali Radix in treatment of kidney disease[J]. Chin Tradit Herb Drugs (中草药), 2018, 49:3419-3424.
[10] Ye LB, Mao W, Li C, et al. Evaluation of adriamycin-induced nephropathy model with syndrome of deficiency of kidney yang[J]. Chin J Tradit Chin Med Pharm (中华中医药杂志), 2016, 31:2146-2149.
[11] Fu D, Xia HY. Treatment of nephrotic syndrome with integrated traditional Chinese and Western medicine[J]. J Changchun Univ Tradit Chin Med (长春中医药大学学报), 2013, 29:242-244.
[12] Xu F, Zhang Y, Xiao S, et al. Absorption and metabolism of Astragali Radix decoction:in silico, in vitro, and a case study in vivo[J]. Drug Metab Dispos, 2006, 34:913-924.
[13] Wang N, Wei R, Li Q, et al. Protective effects of astragaloside in rats with adriamycin nephropathy and underlying mechanism[J]. Chin J Nat Med, 2016, 14:270-277.
[14] Lei X, Zhang L, Li Z, et al. Astragaloside IV/lncRNA-TUG1/TRAF5 signaling pathway participates in podocyte apoptosis of diabetic nephropathy rats[J]. Drug Des Dev Ther, 2018, 12:2785-2793.
[15] Zhang WN, LiAP, Qi YS, et al. Metabolomics coupled with system pharmacology reveal the protective effect of total flavonoids of Astragali Radix against adriamycin-induced rat nephropathy model[J]. J Pharm Biomed Anal, 2018, 158:128-136.
[16] Yu WC, Wang SJ, Ji XM, et al. Influence of water decoction of Huang qi on immune function in rats with deficiency cold pattern:a study based on transcriptome sequencing technique[J]. J Beijing Univ Tradit Chin Med (北京中医药大学学报), 2018, 41:759-770.
[17] Zhang X, Song JY, Hu YL, et al. Research progress of the regulation on active compound biosynthesis by the bHLH transcription factors in plants[J]. Acta Pharm Sin (药学学报), 2014, 49:435-442.
[18] Li AP, Zhang WN, Qin XM. Optimization of adriamycininduced nephropathy rat model[J]. Chin Tradit Herb Drugs (中草药), 2018, 49:151-159.
[19] Martin M. Cutadapt removes adapter sequences from highthroughput sequencing reads[J]. Embnet J, 2011, 17:10-12.
[20] Bourgeau T, Augé J, Friedman T. TopHat:supporting experiments through measurement infrastructure federation[C]//International Conference on Testbeds and Research Infrastructures. Berlin:Springer, 2011:542-557.
[21] Anders S, Pyl PT, Huber W. HTSeq-a Python framework to work with high-throughput sequencing data[J]. Bioinformatics,2015, 31:166-169.
[22] Trapnell C, Williams BA, Pertea G, et al. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J]. Nat Biotechnol, 2010, 28:511-515.
[23] Ernst J, Bar-Joseph Z. STEM:a tool for the analysis of short time series gene expression data[J]. BMC bioinf, 2006, 7:191.
[24] Rozen S, Skaletsky H. Primer3 on the WWW for general users and for biologist programmers[J]. Methods Mol Biol, 2000, 132:365-386.
[25] Prasad R, Hadjidemetriou I, Maharaj A, et al. Sphingosine-1-phosphate lyase mutations cause primary adrenal insufficiency and steroid-resistant nephrotic syndrome[J]. J Clin Invest, 2017, 127:942-953.
[26] Liang CL, Wu JB, Lai JM, et al. Protection effect of Zhen-WuTang on adriamycin-induced nephrotic syndrome via inhibiting oxidative lesions and inflammation damage[J]. Evid Based Complement Alternat Med, 2014, 2014:131604.
[27] Liu Y, Su L, Lin Q, et al. Induction of C-Mip by IL-17 plays an important role in adriamycin-induced podocyte damage[J]. Cell Physiol Biochem, 2015, 36:1274-1290.
[28] Sonneveld R, Hoenderop JG, Isidori AM, et al. Sildenafil prevents podocyte injury via PPAR-γ-mediated TRPC6 inhibition[J]. J Am Soc Nephrol, 2017, 28:1491-1505.
[29] Pozzi A. PI3-kinase and TGF-β in glomerular nephropathy:which comes first?[J]. Kidney Int, 2012, 82:507-509.
[30] Wang W, Li C, Yang T. Protection of nitro-fatty acid against kidney diseases[J]. Am J Physiol Renal Physiol, 2015, 310:F697-F704.
[31] Nicolaou N, Margadant C, Kevelam SH, et al. Gain of glycosylation in integrin α3 causes lung disease and nephrotic syndrome[J]. J Clin Invest, 2012, 122:4375-4387.
[32] Gui DK. Celluar and Molecular Mechanism of Aqueous Extract of Astragali Radix on Renal Water and Sodium Excretion and Its Renal Protective Effects (黄芪水提物促肾脏水钠排泄及肾脏保护作用的细胞分子机制研究)[D]. Shanghai:Fudan University, 2007.
[33] Agrawal S, Zaritsky JJ, Fornoni A, et al. Dyslipidaemia in nephrotic syndrome:mechanisms and treatment[J]. Nat Rev Nephrol, 2017, 14:57-70.
[34] Song SB. PCSK9-new target of hypercholesterolemia[J]. Med Recapitul (医学综述), 2016, 22:1080-1084.
[35] Ling B, Aziz C, Alcorn J. Systematic evaluation of key L-carnitine homeostasis mechanisms during postnatal development in rat[J]. Nutr Metab, 2012, 9:66.
[36] Van Weeghel M, Abdurrachim D, Nederlof R, et al. Increased cardiac fatty acid oxidation in a mouse model with decreased malonyl-CoA sensitivity of CPT1B[J]. Cardiovasc Res, 2018, 114:1324-1334.
[37] Zhang WN, Li AP, Liu SB, et al. Evaluation of injury degree of rat nephropathy model induced by doxorubicin based on metabolomics[J]. Chin Tradit Herb Drugs (中草药), 2018, 49:360-367.
[38] Gao JR, Qin XJ, Jiang H, et al. The effects of Qi Teng Xiao Zhuo granules, traditional Chinese medicine, on the expression of genes in chronic glomerulonephritis rats[J]. J Ethnopharmacol, 2016, 193:140-149.
[39] Zhu Q, Diao FY, Liu JY. Orphan nuclear receptor NR4A1 regulating apoptosis via mitochondria-related mechanisms[J]. J Interent Rep Health/Fam (国际生殖健康/计划生育杂志), 2017, 36:488-491.
[40] Sun H, Wang W, Han P, et al. Astragaloside IV ameliorates renal injury in db/db mice[J]. Sci Rep, 2016, 6:32545.