Orai1体外干预对宫颈息肉炎症反应的作用

Effect of Orai1 protein intervention on inflammatory reactions in cervical polyps in vitro

目的探讨Orai1的体外干预对宫颈息肉(CP)中炎症反应的作用。方法收集复旦大学附属华山医院北院病理科2016年至2019年期间40例CP患者的息肉组织,用器官培养技术进行体外培养,然后将2-氨基乙基联苯基硼酸酯(2-APB)作为体外干预手段,以免疫组织化学染色方法检测2-APB干预前后CP中Orai1蛋白的表达情况,接着用酶联免疫吸附试验(ELISA)和实时定量反转录聚合酶链反应(RT-PCR)分别检测2-APB干预前后CP组织中Orai1蛋白及其mRNA的含量,最后再用上述方法检测CP组织中白介素(IL)-1β、肿瘤坏死因子(TNF)-α及其mRNA的浓度。结果免疫组织化学染色显示CP标本中表达Orai1蛋白,而且2-APB干预后该蛋白的染色评分减少;ELISA法和实时定量RT-PCR法检测结果表明,2-APB干预后CP中Orai1蛋白及其mRNA的含量均较干预前减少(t值分别为5.76、10.85,均P<0.05),炎症介质IL-1β、TNF-α及其mRNA的浓度也显著低于干预前(t值分别为15.28、6.35、7.10、17.74,均P<0.05)。结论 Orai1的体外干预可以抑制CP中的炎症反应。

Objective To explore effect of Orai1 protein intervention on inflammatory reactions in cervical polyps(CP) in vitro.Methods CP tissues that were preserved in Department of Pathology of our hospital from 40 patients who were diagnosed and treated in our hospital from years 2016 to 2019 were obtained and cultured in vitro in organ culture technique.2-Aminoethoxydiphenyl borate(2-APB) was selected as intervention agent in vitro.Immunohistochemical staining was used to detect expression of Orai1 protein in CP tissues before and after the intervention.Then enzyme-linked immunosorbent assay(ELISA) and real-time quantitative reverse transcription-polymerase chain reaction(RTq-RT-PCR) were used to detect concentrations of Orai1 protein and its mRNA in CP tissues before and after administration of 2-APB.Finally,the contents of inflammatory mediators such as interleukin-1β(IL-1β) and tumor necrosis factor α(TNF-α) and their mRNAs in CP tissues were also detected in above methods.Results Orai1 protein was expressed in CP tissues cultured in vitro,and its score in immunohistochemical staining was decreased after 2-APB administration.The detection results by ELISA and RTq-RT-PCR showed that Orai1 protein(t=5.76,P<0.000 1) and its mRNA(t=10.85,P <0.000 1) in CP tissues in vitro were reduced after the intervention by 2-APB,and also,the concentrations of inflammatory mediators IL-1β(t=15.28,P <0.000 1) and TNF-α(t=6.35,P<0.000 1) and their mRNAs(t=7.10,P<0.000 1;t=17.74,P<0.000 1) were significantly lower than those before the invention.Conclusion The intervention on Orai1 protein may inhibit inflammatory reactions in CP in vitro.