汉黄芩苷通过调控MAPK信号通路减轻高血压所致肾脏损伤的机制研究  

Wogonoside Attenuates Hypertension-Induced Renal Injury Through Modulation of the MAPK Signaling Pathway:A Mechanism Study

作　　者：郭智 谢意 刘红纾 肖珺丹 陈榕基 吴美珠 彭军 沈阿灵 GUO Zhi;XIE Yi;LIU Hongshu;XIAO Jundan;CHEN Rongji;WU Meizhu;PENG Jun;SHEN Aling(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中西医结合研究院,福州350122 [2]福建中医药大学中西医结合学院,福州350122 [3]福建省中西医结合老年性疾病重点实验室,福州350122

出　　处：《四川大学学报(医学版)》2025年第1期41-50,共10页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.U22A20372、No.82074363);中华中医药学会2021-2023年度青年人才托举工程项目(No.2021-QNRC2-B19);福建中医药大学青年科研拔尖人才(No.XQB202202);福建省卫生健康中青年领军人才研修培养项目(闽卫人函[2023]2841)资助。

摘　　要：目的通过高通量基因表达数据库(GEO)和网络药理学方法探讨汉黄芩苷对高血压所致肾损伤的潜在治疗作用、靶点及通路,并通过体内外实验验证汉黄芩苷干预对自发性高血压大鼠(spontaneously hypertensive rats,SHR)肾脏组织和血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)刺激大鼠肾细胞NRK-52E凋亡的影响及对相关通路的调控作用。方法通过GEO数据库、网络药理学分析汉黄芩苷治疗高血压性肾病的关键治疗靶点,STRING数据库分析治疗靶点间的互作关系,基因本体(GO)数据库分析相关生物学功能,京都基因与基因组百科全书(KEGG)数据库富集潜在信号通路。将SHR大鼠随机分组并分别灌胃给予低、中、高剂量(0.075、0.75和7.5 mg/kg)汉黄芩苷,干预10周后,通过HE染色检测肾脏组织形态的改变,ELISA检测血清中炎症因子肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6的表达,TUNEL染色检测细胞凋亡率,Western blot检测Bax、Bcl-2、cleaved caspase-3、caspase-3表达及细胞外信号调节激酶(extracellular regulated protein kinases,ERK)、p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)的磷酸化和总蛋白的表达。构建AngⅡ刺激的NRK-52E细胞模型,采用不同剂量汉黄芩苷(25、50、100μmol/L)干预细胞24 h,通过Annexin V染色检测细胞凋亡率以及Western blot验证上述凋亡相关蛋白及通路相关蛋白的表达。结果通过数据集GSE41453分析,发现与WKY组相比,SHR组肾脏组织中有11673个基因上调和5902个基因下调,通过多个数据库获取汉黄芩苷371个潜在作用靶点,从而获取98个交集靶点,进一步筛选得到45个核心治疗靶点,其中包括TNF、CASP3等靶点。通过GO数据库分析,凋亡过程的负向调控等过程被显著富集。通过KEGG数据库富集分析发现,凋亡通路、IL-17信号通路和MAPK信号通路等通路被显著富集。汉黄芩苷干预可有效抑制SHR大�Objective To investigate the potential therapeutic effects,targets,and pathways of wogonoside in hypertension-induced renal injury using the Gene Expression Omnibus(GEO)database and network pharmacology,and to validate the effects of wogonoside intervention on the renal tissues of spontaneously hypertensive rats(SHR),angiotensinⅡ(AngⅡ)-stimulated NRK-52E cell apoptosis,and the regulation of relevant pathways through in vivo and in vitro experiments.Methods GEO dataset and network pharmacology analyses were performed to investigate the key therapeutic targets of wogonoside for hypertensive nephropathy.The STRING database was used to analyze proteinprotein interactions.Biological functions were annotated via Gene Ontology(GO),and the potential signaling pathways were enriched using the Kyoto Encyclopedia of Genes and Genomes(KEGG).SHR were randomly divided into groups and given low,medium,or high doses of wogonoside(0.075,0.75,and 7.5 mg/kg)via gastric gavage for 10 weeks.Morphological changes in the kidney tissue were assessed by hematoxylin-eosin(HE)staining.Serum levels of inflammatory cytokines,including tumor necrosis factorα(TNF-α),interleukin(IL)-1β,and IL-6,were measured using ELISA.Apoptosis rates were evaluated by TUNEL staining,and Western blot was performed to determine the expression of Bax,Bcl-2,cleaved caspase-3,and caspase-3,and the expression of phosphorylated and total extracellular signalregulated kinases(ERK)and p38 mitogen-activated protein kinase(MAPK)proteins.An in vitro model of AngⅡ-stimulated NRK-52E cells was constructed and was treated with wogonoside at different concentrations(25,50,or 100μmol/L)for 24 h.The apoptosis rates were then assessed by Annexin V staining,and Western blot was performed to validate the expression of apoptosis-related and pathway-associated proteins.Results Analysis of dataset GSE41453 revealed 11673 upregulated and 5902 downregulated genes in the renal tissues of SHR compared to the Wistar Kyoto(WKY)rats,or the WKY control group.Through the analysis o

关 键 词：高通量基因表达数据库 网络药理学 汉黄芩苷 高血压性肾病 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]