基于生物信息学分析雌激素对软骨细胞分泌炎症因子表达谱的影响  

Analysis of the Effect of Estrogen on the Expression Profile of Inflammatory Factors Secreted by Chondrocytes Based on Bioinformatics

作　　者：李永胜 黄泽祈[2,3] 赵喆 邓志钦[3] 邓桢翰 李文翠 LI Yongsheng;HUANG Zeqi;ZHAO Zhe;DENG Zhiqin;DENG Zhenhan;LI Wencui(Shantou University Medical College,Guangdong Shantou 515000;Shenzhen University Medical school,Guangdong Shenzhen 518060;Shenzhen Second People’s Hospital,Guangdong Shenzhen 518035;The First Affiliated Hospital of Wenzhou Medical University,Zhejiang Wenzhou 325000)

机构地区：[1]汕头大学医学院,广东汕头515000 [2]深圳大学医学部,广东深圳518060 [3]深圳市第二人民医院,广东深圳518035 [4]温州医科大学附属第一医院,浙江温州325000

出　　处：《深圳中西医结合杂志》2024年第10期1-6,I0002,I0003,共8页Shenzhen Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金面上项目(82172465);中国高校产学研创新基金(2021JH037);广东省临床重点学科骨科(2000005);广东省自然科学基金项目(2021A1515010706,2023A1515010102);深圳市重点医学学科建设基金(SZXK025);深圳市科技计划国际合作项目(GJHZ20210705142007023);深圳市医疗卫生“三名工程”项目(SZSM202311008)。

摘　　要：目的:探讨雌激素介导软骨细胞分泌炎症因子的机制。方法:收集深圳市第二人民医院收治的股骨颈骨折患者,从髋关节置换术中无菌取出的股骨头中,分离培养关节软骨细胞。空白对照组仅加入软骨细胞培养液,其他观察分组除软骨细胞培养液,分别加入雌二醇(E2)、雌激素受体α(ERα)、雌激素受体β(ERβ)和雌激素受体阻断剂氟维司群(Ful),均培养48 h,与空白对照组进行比较;通过GSH-INF-3芯片分析软骨细胞分泌的炎症因子,在DAVID网站(https://david.ncifcrf.gov/)进行信号通路富集基因本体(GO)和京都基因组与基因组百科全书(KEGG)分析,阐明雌激素调控软骨细胞表达关键基因与分子标记。结果:观察组中各组与空白对照组比较,均鉴定出差异蛋白表达,通过蛋白质谱分析筛选得到目的基因,当雌激素介导软骨细胞分泌验证因子时,上调的基因包括:白细胞介素(IL)-1α、IL-5、IL-11、细胞间黏附分子-1(ICAM-1)、肿瘤坏死因子受体I(TNF-RI)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)、IL-12P70、组织金属蛋白酶抑制物(TIMP)-1和巨噬细胞炎性蛋白(MIP)-1β;下调的基因包括B淋巴细胞趋化因子(BLC)、单核细胞集落刺激因子(MCSF)、单核细胞趋化蛋白-1(MCP-1)、IL-17、IL-2、IL-4、TNF-α和IL-8;GO分析示差异基因多富集于细胞因子介导的信号通路、内质网的内腔、信号受体激活剂活性和受体配体活性。KEGG分析示以细胞因子活性、细胞因子-细胞因子受体相互作用和类风湿性关节炎通路参与雌激素激活软骨细胞炎症因子。结论:雌激素主要通过细胞因子活性和细胞因子-细胞因子受体相互作用通路,上调IL-5、IL-11、TNF-RⅠ、ICAM-1和IL-1α等软骨细胞分泌因子的关键基因,其可能成为骨性关节炎(OA)的潜在治疗靶点。Objective To investigate the mechanism of estrogen mediated chondrocyte secretion of inflammatory cytokines.Methods Patients with femoral neck fracture admitted to Shenzhen Second People's Hospital were collected,and the femoral head was sterile removed during hip replacement,and the articular chondrocytes were isolated and cultured.Only chondrocyte culture medium was added to the blank control group.In addition to chondrocyte culture medium,estradiol(E2),estrogen receptorα(ERα),estrogen receptorβ(ERβ)and estrogen receptor blocker fluvestrant(Ful)were added to the other observation groups,and cultured for 48 h,compared with the blank control group.By analyzing GSH-IN-3 chip chondrocytes secretion of the inflammatory factor,enrichment of signaling pathways in DAVID website(https://david.ncifcrf.gov/)gene ontology(GO)and the Kyoto encyclopedia genome and genomes(KEGG)analysis,to elucidate the key genes and molecular markers of estrogen regulation of chondrocyte expression.Results Compared with the blank control group,different protein expressions were identified in all groups in the observation group.The target genes were screened through protein profiling.When estrogen mediated chondrocyte secretion of validation factors,the up-regulated genes included:interleukin(IL)-1α,IL-5,IL-11,intercellular adhesion molecule-1(ICAM-1),tumor necrosis factor(TNF)-RI,granulocyte-macrophage colony-stimulating factor(GM-CSF),IL-12P70,tissue inhibitor of metalloproteinase(TIMP)-1,and macrophage inflammatory protein(MIP)-1β.Down-regulated genes included B lymphocyte chemokine(BLC),monocyte colony stimulating factor(MCSF),monocyte chemokine protein-1(MCP-1),IL-17,IL-2,IL-4,TNF-αand IL-8.GO analysis showed that differential genes were mainly concentrated in cytokine-mediated signaling pathways,endoplasmic reticulum lumen,signaling receptor activator activity and receptor ligand activity.KEGG analysis showed that cytokine activity,cytokine-cytokine receptor interactions,and the rheumatoid arthritis pathway were involved in estro

关 键 词：骨性关节炎 软骨细胞 雌激素 炎症因子 生物信息学 

分 类 号：R684.3[医药卫生—骨科学]