汉防己甲素对人眼Tenon囊成纤维细胞表达细胞凋亡蛋白和转化生长因子的影响

背景汉防己甲素（Tet）对体外培养的人眼Tenon囊成纤维细胞（TCFs）有抑制效应，但其作用机制尚不完全明确。目的观察Tet对体外培养的人眼TCFs中bax、bel-2、转化生长因子β2（TGF—β2）蛋白表达的影响，探讨其可能的作用机制。方法收集健康供体眼球巩膜组织，用组织块培养法培养、分离TCFs，并进行传代，将第3代细胞以1×10^5个／ml的密度接种于培养板，采用光学显微镜和免疫组织化学法对培养细胞进行鉴定。培养24h后，用培养液稀释Tet至浓度为1×10^-5mol／L，并加入培养孔中，对照组用1640培养液，继续培养24h后冲洗固定，采用TUNEL法检测细胞的凋亡情况；采用免疫组织化学法检测bax、bcl-2、TGF—β2蛋白在TCFs中的表达。结果培养的细胞形态学呈现成纤维细胞的特征，vimentin染色呈阳性反应。Tet组出现大量的TUNEL阳性细胞，而对照组TCFs未见明显凋亡细胞核出现。免疫组织化学检测表明，Tet组TCFs中bax、bcl-2、TGFβ2蛋白的表达率（A值）分别为0．577±0．009、0．430±0．012、0．341±0．017，对照组分别为0．320±0．015、0．819±0．021、0．624±0．014，Tet组bax的表达明显升高，bel-2和TGFβ2的表达明显下降，差异均有统计学意义（t=33．277、-35．356、-28．093，P〈0．01）。结论Tet通过上调人眼Tenon囊成纤维细胞中bax的表达，下调bcl-2、TGF-β2的表达，从而抑制TCFs的增生。

The Expression of Apoptosis-Related Genes Bcl-2 and Bax Protein and Apoptosis Positivity in Cervical Carcinoma during Irradiation

To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.