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目的探讨天然药物姜黄素对晶状体上皮细胞(LEC)凋亡的诱导效应及其机制。方法通过透射电镜观察姜黄素对体外培养的牛LEC超微结构的影响;采用流式细胞术(FCM)观察姜黄素对LEC的DNA含量及线粒体跨膜电位(AΨm)的影响。结果透射电镜下可观察到姜黄素组LEC呈现核染色质凝聚、固缩、边集、核碎裂等典型的细胞凋亡形态学改变。经FCM检测显示,姜黄素组作用早期(24 h)LEC细胞核的DNA量没有改变,在作用的中、晚期(48、72 h)细胞核内DNA呈现明显的下降趋势,DNA含量显著低于空白对照组(P<0．01);而姜黄素作用早期线粒△Ψm已经开始下降,8 h时△Ψm下降的细胞数占(4．13±0．52)％;作用72 h后达高峰,占(46．91±3．71)％。各期与正常对照组的差异均有统计学意义(P<0．01)。结论姜黄素可通过细胞核和细胞质两种途径诱导LEC凋亡。诱导LEC凋亡可能是姜黄素减少晶状体后囊膜混浊的细胞和分子机制,姜黄素可能成为一种新型的、高效低毒的防治后发性白内障的药物。
Objective To investigate the inducing effect of natural drug curcumin on apoptosis of lens epithelial cells (LECs) and its mechanism. Methods The effects of curcumin on the ultrastructure of bovine LEC cultured in vitro were observed by transmission electron microscopy. The effects of curcumin on DNA content of LEC and mitochondrial transmembrane potential (AΨm) were observed by flow cytometry (FCM). RESULTS: Under TEM, the morphological changes of apoptotic cells were observed in the LEC of the curcumin group, which showed nuclear chromatin condensation, pyknosis, edge set, and nuclear fragmentation. The FCM assay showed that the amount of DNA in the LEC nuclei did not change in the early (24 h) phase of the curcumin group. DNA in the nuclei of the mid- and late-phase (48, 72 h) showed a declining trend, and the DNA content was significantly lower than that in the control group. In the group (P<0.01), the early mitochondrial △Ψm of curcumin had begun to decline, and the number of △Ψm decreased (4.13±0.52)% at 8 h; after 72 h, peaked. It accounts for (46.91±3.71)%. The difference between each period and the normal control group was statistically significant (P<0.01). Conclusion Curcumin can induce LEC apoptosis through both nuclear and cytoplasmic pathways. The induction of LEC apoptosis may be the cellular and molecular mechanisms by which curcumin reduces lens opacification, and curcumin may become a novel, highly effective, and low toxic drug for the prevention and treatment of later-onset cataracts.