CXCL1/2/10介导的免疫炎症与草酸钙结石形成的关系

Relationship between CXCL1/2/10 mediated immune inflammation and calcium oxalate stone formation

目的:探讨CXC趋化因子配体1、2、10(CXCL1/2/10)等炎症诱导因子的差异分布及其介导的免疫炎症与草酸钙结石形成的关系。方法:首先在GEO数据库中进行数据检索,获得高钙尿大鼠的mRNA测序矩阵后,对其进行差异基因分析;对获得的差异基因进行GO与KEGG的功能富集分析;对获得的差异基因进行蛋白互作分析(PPI)进而筛选关键分子;通过饲喂1%乙二醇与灌胃1%的氯化铵构建草酸钙结石的大鼠模型,并进行HE染色验证模型;在草酸钙结石大鼠的肾组织切片中进行免疫组化实验,以验证关键分子的表达水平。结果:基于GEO数据共获得195个差异表达基因,其中上调146,下调49(P<0.05,log_(2)FC<-1或log_(2)FC>1);基于Metascape数据库得出的GO与KEGG分析显示,基因富集在GO:0032496:对脂多糖的反应、GO:0010942:细胞死亡的正调控、GO:0002544:慢性炎症反应、GO:0006954:炎症反应、GO:0072593:活性氧代谢过程等代谢通路中;PPI蛋白互作分析结合功能富集分析最终确定5个关键基因,即CXCL1、CXCL2、CXCL10、IL6、IL1B;HE染色结果显示草酸钙结石大鼠模型构建成功,且免疫组化显示5个关键分子在造模组中均升高,差异具有统计学意义(P<0.05),其中CXCL1/2/10表达明显升高(P<0.001)。结论:基于生物信息学分析与实验验证,CXCL1/2/10在草酸钙结石大鼠肾组织中表达明显升高,可能与免疫炎症相关,提示CXCL1/2/10可成为缓解草酸钙结石患者肾组织免疫炎症进而达到治疗效果的潜在分子靶点。

Objective:To investigate the differential distribution of CXC(CXCL1/2/10)and other inflammatory inducing factors and the relationship between immune inflammation mediated by CXCL1/2/10 and calcium oxalate stone formation.Methods:Firstly,we searched the GEO database to obtain the mRNA sequencing matrix of hypercalciuria rats to analyze the differential genes;After that,the functional enrichment of GO and KEGG was analyzed;Protein interaction analysis(PPI)was used to screen the key molecules;The rat model of calcium oxalate calculus was established by feeding 1%ethylene glycol and intragastric administration of 1%ammonium chloride,and the model was verified by HE staining;In order to verify the expression level of the key molecules,immunohistochemistry was performed in renal tissue sections of calcium oxalate stone rats.Results:A total of 195 differentially expressed genes were obtained based on GEO data,including 146 up-regulated genes and 49 down regulated genes(P<0.05,log_(2)FC<-1 or log_(2)FC>1);Based on Metascape(http:∥metascape.org/)website,GO and KEGG analysis results showed that genes were enriched in some terms such as GO:0032496:response to lipopolysaccharide,GO:0010942:response to positive regulation of cell death,GO:0002544:response to chronic inflammatory,GO:0006954:response to inflammatory,GO:0072593:response to metabolic process of reactive oxygen species and other metabolic pathways;PPI protein interaction analysis combined with functional enrichment analysis finally identified five key genes:CXCL1,CXCL2,CXCL10,IL6,IL1B;HE staining results showed that the rat model of calcium oxalate stone was successfully constructed,and the results of immunohistochemistry showed that the five key molecules were increased in the model group,the difference was statistically significant(P<0.05),and the expression of CXCL1/2/10 was significantly increased(P<0.001).Conclusion:Based on bioinformatics analysis and experimental verification,this study has found that CXCL1/2/10 is significantly increased in renal tissue of rats with calcium oxalate calculi,which may be related to immune inflammation,suggesting that CXCL1/2/10 may be a potential molecular target for alleviating renal immune inflammation in patients with calcium oxalate calculi and achieving therapeutic effect.