转化生长因子β_2诱导人小梁细胞凋亡的体外研究

Apoptosis of human trabecular meshwork cells induced by transforming growth factor-β_2 in vitro

目的 观察转化生长因子 β2 (transforminggrowthfactor β2 ,TGF β2 )是否能诱导体外培养人眼小梁细胞 (humantrabecularmeshworkcells,HTMC)发生凋亡。方法 将体外培养的第 3～ 5代人眼小梁细胞分为对照组和实验组 ,实验各组分别经 0 .32、1.0 0、3.2μg·L-1TGF β2 作用 4 8h后 ,分别用透射电镜、TUNEL法和流式细胞术检测细胞凋亡的诱导情况。结果 透射电镜观察发现凋亡细胞典型的形态学变化———核染色质凝集边集 ,凋亡小体形成等。TUNEL法检出发生DNA片段化的HTMC。经流式细胞术定量分析 ,不同浓度TGF β2 处理的HTMC凋亡细胞百分数分别为 ( 2 .79± 0 .4 4 ) %、( 4 .4 3±1.17) %、( 9.6 0± 2 .0 5 ) % ,其中 1、3.2 μg·L-1TGF β2 处理组与对照组 ( 1.4 1± 0 .34) %相比分别有显著差异和极显著差异。结论 TGF β2 能诱导体外培养HTMC发生凋亡 ,推测其可能参与了原发性开角型青光眼患者和正常人衰老过程中HTMC数目的减少。

Objective To investigate whether transforming growth factor β 2(TGF β 2) could induce apoptosis of human trabecular meshwork cells in vitro. Methods Cultrued 3～5 passage human trabecular meshwork cells were divided into control group and experimental group,and were treated with 0 (control),0.32,1.00 and 3.2 μg·L -1 TGF β 2 for 48 hours, apoptosis of human trabecular meshwork cells were examined by transmisson electron microscope,TUNEL technique and flow cytometry.Results Characteristic morphologic changes of apoptotic cells were observed under transmission electron microscope with chromatin condensation and margination as well as formation of apoptotic bodies.DNA fragmentation of human trabecular meshwork cells were found by TUNEL technique.Quantitative analysis of flow cytometry showed that percentages of apoptotic human trabecular meshwork cells were (2.79±0.44)%, (4.43±1.17)% and (9.60±2.05)% with different concentrations of TGF β 2, there were significant difference between experimental groups treated with 1μg·L -1 ( P <0.05),3.2μg·L -1 ( P <0.01) TGF β 2 and ( 1.41 ± 0.34 )% of control group.Conclusion TGF β 2 can induce apoptosis of human trabecular meshwork cells in vitro. It may be involved in the decrease of trabecular meshwork cells in patients of primary open angle glaucoma and aging of normal people.