摘要
通过数据挖掘和动物实验探讨大黄素治疗脓毒症相关急性肾损伤(sepsis-associated acute kidney injury,SA-AKI)的分子机制。从GEO数据库筛选SA-AKI发病过程的关键基因,进行GO和KEGG富集分析,然后利用STRING构建PPI网络,采用Cytoscape中的MCODE构建核心靶标、炎性信号通路互作网络。Wistar大鼠构建三组模型:假手术组(Sham)、脓毒症组(CLP)和药物组(CLP+EMO),运用HE染色法、脲酶法、肌氨酸氧化酶法、硫代巴比妥酸法、ELISA法和Western blot法等实验方法进行检测。结果显示,共获得2801个SA-AKI发病过程的关键靶点。GO生物过程分析提示这些靶点的生物学功能主要涉及膜信号转导、血管调节和伤口愈合。KEGG通路富集分析显示,TNF、IL-17、PI3K-Akt、NF-κB和MAPK信号通路是富集与炎症相关的信号通路。三组模型6 d内死亡率无差异(P>0.05)。与Sham组相比,CLP组肾小球大小不一,有萎缩和分裂现象,肾小管扩张,空泡样改变多见,高倍镜下可见炎性细胞浸润;血液中Scr、BUN和MDA水平显著上升(P<0.001);细胞因子TNF-α、IL-17和IFN-γ水平显著升高(P<0.001);且肾组织中TNF-α、IL-17A、TRAF6和NF-κB信号通路蛋白表达上调(P<0.05)。与CLP组比较,CLP+EMO组空泡样改变降低,高倍镜下炎性细胞浸润减少;血液中测定的上述指标与肾组织测定的蛋白含量表达均呈现不同程度的降低(P<0.01、P<0.001和P<0.05)。综上,大黄素可以改善SA-AKI肾功能水平并减轻其炎症反应,可能与下调IL-17/NF-κB信号通路和TNF/NF-κB信号通路有关。
To investigate the molecular mechanism of emodin in the treatment of sepsis-associated acute kidney injury(SA-AKI)by data mining and animal experiments.Critical genes in the pathogenesis of SA-AKI were screened from the GEO database for GO and KEGG enrichment analysis,and then PPI networks were constructed using STRING,and core target,inflammatory signaling pathway interaction networks were constructed using MCODE in Cytoscape.Wistar rats were used to construct three groups of models:sham operation group(Sham),sepsis group(CLP)and drug group(CLP+EMO).HE staining,urease,sarcosine oxidase,thiobarbituric acid,ELISA and Western blot were used to detect the three groups of models.The results showed that a total of 2801 key targets for the pathogenesis of SA-AKI were obtained.GO biological process analysis suggests that the biological functions of these targets mainly involve membrane signal transduction,vascular regulation,and wound healing.KEGG pathway enrichment analysis showed that TNF,IL-17,PI3K-Akt,NF-κB and MAPK signaling pathways were enriched signaling pathways associated with inflammation.There was no difference in mortality within 6 days among the three groups(P>0.05).Compared with the sham group,glomeruli in the CLP group varied in size,with atrophy and division,tubular dilatation,vacuole-like changes were more common,and inflammatory cell infiltration was observed at high magnification;Scr,BUN,and MDA levels in the blood were significantly increased(P<0.001);cytokines TNF-α,IL-17,and IFN-γlevels were significantly increased(P<0.001);and TNF-α,IL-17A,TRAF6,and NF-κB signaling pathway protein expression was up-regulated in the renal tissue(P<0.05).Compared with the CLP group,vacuolated changes were decreased and inflammatory cell infiltration was decreased in the CLP+EMO group at high magnification;the above indexes measured in blood and the protein content expression measured in renal tissue showed different degrees of decrease(P<0.01,P<0.001 and P<0.05).In summary,emodin can improve renal function levels and reduce the inflammatory response in SA-AKI,which may be related to the down-regulation of IL-17/NF-κB signaling pathway and TNF/NF-κB signaling pathway.
作者
欧贤
周金瑶
孙饶
王亚萍
吴祯祥
赵凯奇
汪华学
OU Xian;ZHOU Jin-yao;SUN Rao;WANG Ya-ping;WU Zhen-xiang;ZHAO Kai-qi;WANG Hua-xue(Department of Critical Care Medicine of the First Affiliated Hospital of Bengbu Medical College;Institute of Emergency and Critical Care Medicine of the First Affiliated Hospital of Bengbu Medical College;Bengbu Medical College,Bengbu 233000,China)
出处
《天然产物研究与开发》
CAS
CSCD
2023年第8期1422-1430,1401,共10页
Natural Product Research and Development
基金
安徽省重点研究和开发计划(1804h08020256)
蚌埠医学院研究生创新计划(Byycx21099)
蚌埠医学院大学生创新培训项目(bydc2021066)。
作者简介
通信作者:汪华学,Tel:86-013965282230,E-mail:huaxuew2010@163.com。
