低温暴露诱导卵巢组织TNF-α表达水平上调的信号转导机制

Signal transduction mechanism involved in up regulation of TNF-α expression in ovarian tissue induced by cold exposure

目的探讨低温暴露对卵巢组织炎性因子表达的影响及相关分子机制。方法将6~8周龄雌性小鼠随机分为对照组以及低温暴露组,暴露后取卵巢组织,通过RT-PCR方法检测炎性因子表达水平,Western印迹法检测炎性反应相关蛋白的表达变化及活化情况。体外培养SKOV3细胞,进行温和冷暴露(35℃),通过RT-PCR和Western印迹法检测炎性因子和炎性反应相关蛋白的表达水平及活化情况。结果低温暴露后卵巢的组织局部出现肿瘤坏死因子(TNF-α)表达水平的显著上调,炎性转录因子NF-κB p65活化。体外培养的SKOV3卵巢癌细胞经温和冷刺激后,同样能观察到TNF-α表达水平的显著上调和炎性转录因子NF-κB p65的诱导活化;在SKOV3细胞中转染NF-κB p65 siRNA后,可显著抑制TNF-α的诱导表达。结论低温暴露通过诱导卵巢组织NF-κB p65活化介导炎性因子TNF-α上调及炎症反应。

Objective To investigate the effect of low temperature exposure on the expression of inflammatory factors in ovarian tissues and the related signal transduction mechanism.Methods Female mice aged 6-8 weeks were randomly divided into two groups:one group was left untreated while the other group was subjected to cold exposure.The expression levels of inflammatory factors and signaling molecules involved in inflammatory responses were detected in ovarian tissues of mice from both groups by RT-PCR and Western blotting assays,respectively.SKOV3 cells were cultured under normal(37℃) or mild cold(35℃) conditions.The inflammatory factors and inflammation related proteins were detected by RT-PCR and Western blotting assays.Results The tumor necrosis factor-α(TNF-α) level was up-regulated and NF-κB p65 was activated in the ovarian tissues after cold exposure.In addition,an increase in TNF-α expression and activation of NF-κB p65 were also observed in SKOV3 cells cultured under 35℃.Moreover,up-regulation of TNF-α expression induced by cold exposure was inhibited in SKOV3 cells after transfection of NF-κB p65 siRNA.Conclusion Cold exposure induces up-regulation of TNF-α expression and inflammatory responses in ovarian tissues by activating NF-κB p65.