从受体角度探讨黄芩苷对肺炎衣原体感染细胞的干预机理

Exploring the Mechanism of Baicalin in Influencing Cells Infected with Chlamydia Pneumoniae by Observing Its Effect on Receptors

[目的]观察黄芩苷对肺炎衣原体诱导的TOLL上受体(TLRs)的影响。[方法]体外培养人脐静脉内皮细胞(ECV-304 细胞),待长成单层后加入肺炎衣原体(CPN)为模型组,不加任何刺激者为正常组,每组设3个复孔,6孔板培养2 d后, 用单克隆抗体荧光标记和姬姆萨染色检测肺炎衣原体在ECV-304细胞内的生长情况;用逆转录聚合酶链反应(RT-PCR) 法检测细胞TLR2 mRNA、TLR4 mRNA表达。另取长成单层的ECV-304细胞,加入CPN为模型组,加入CPN+黄芩苷0.48 g/L为黄芩苷高剂量组,加入CPN+0.24 g/L为黄芩苷低剂量组,用流式细胞仪检测ECV-304细胞表面TLR2蛋白表达。[结果]CPN能在ECV-304细胞内生长;该细胞检测不出TLR4 mRNA的表达,存在TLR2 mRNA和TLR2蛋白的高表达。中药黄芩苷高剂量可降低TLR2蛋白的表达。[结论]黄芩苷对CPN所刺激的TLR2高表达具有抑制作用。

[Objective] To observe the effect of Baicalin on the expression of Toll-like receptors (TLRs) induced by chlamydia pneumoniae (CPN). [Methods] The endothelial cells of umbilical veins (ECV-304) was cultured in-vitro in 6-well culture plate (3 double-wells as a group). When a single layer formed, CPN or CPN + baicalin (0.48 g/L or 0.24 g/L) was added, serving as the model group and baicalin groups respectively. The wells without adding agents served as the normal group. After 5 days of culture, the intracellular growth of CPN in ECV-304 was examined by monoclonal antibody fluorescence labeling method and Giemsa stain, and the expression of TLR2 mRNA and TLR4 mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR) in the model group. Protein expression of TLR2 on ECV-304 cell surface in the cultured supernatants was detected with flow cytometer. [Results] The intracellular growth of CPN can be found in ECV-304. The expression of TLR4 mRNA was undetectable, and high expression of TLR2 mRNA and TLR2 protein existed in ECV-304. Baicalin in high doses could lower the expression of TLR2 protein. [Conclusion] Baicalin has an inhibitory effect on the high expression of TLR2 protein in ECV-304 stimulated by CPN.