School of Life Science and Technology, China Pharmaceutical University, Nanjing, China.
Toxicol Lett. 2012 Nov 23;215(1):25-30. doi: 10.1016/j.toxlet.2012.09.017. Epub 2012 Oct 2.
Apoptosis of lens epithelial cell (LEC) plays an important role in cataract formation, and its prevention may be one of the therapeutic strategies in treating cataract. This study used human lens epithelial cell (hLEC) line SRA01/04 to investigate the protective effect and mechanism of phycocyanin on glactose-induced apoptosis in hLEC. hLECs were cultured in D/F(12)-10% FBS medium containing 125mM d-galactose with or without phycocyanin. Cell viability was assessed by methylthiazol tetrazolium (MTT) assay. Cell apoptosis was elevated with Wright-Giemsa staining, AO/EB double staining, and DNA fragmentation assay. Mitochondrial apoptosis-associated molecules and unfolded protein response-associated molecules from cultured SRA01/04 cells were quantified using protein blot analysis. The results demonstrated that phycocyanin suppressed SRA01/04 cells' morphologic changes and apoptosis induced by d-galactose, inhibited the expression and activation of caspase 3, alternated the Bax/Bcl-2 ratio, and down-regulated the level of p53, GRP78, and CHOP in d-galactose-treated SRA01/04 cells. These results suggest that phycocyanin might suppress d-galactose-induced hLEC apoptosis through two pathways: mitochondrial pathway, involving p53 and Bcl-2 family protein expression, and unfolded protein response pathway, involving GRP78 and CHOP expression.
晶状体上皮细胞(LEC)的凋亡在白内障形成中起重要作用,其预防可能是治疗白内障的一种治疗策略。本研究使用人晶状体上皮细胞(hLEC)系 SRA01/04 来研究藻蓝蛋白对 hLEC 葡萄糖诱导凋亡的保护作用和机制。hLEC 在含有 125mM d-半乳糖的 D/F(12)-10% FBS 培养基中或含有藻蓝蛋白的培养基中培养。通过甲基噻唑四唑(MTT)测定法评估细胞活力。通过 Wright-Giemsa 染色、AO/EB 双重染色和 DNA 片段化测定法检测细胞凋亡。使用蛋白印迹分析定量测定培养的 SRA01/04 细胞中的线粒体凋亡相关分子和未折叠蛋白反应相关分子。结果表明,藻蓝蛋白抑制 d-半乳糖诱导的 SRA01/04 细胞形态变化和凋亡,抑制 caspase 3 的表达和激活,改变 Bax/Bcl-2 比值,并下调 d-半乳糖处理的 SRA01/04 细胞中 p53、GRP78 和 CHOP 的水平。这些结果表明,藻蓝蛋白可能通过两条途径抑制 d-半乳糖诱导的 hLEC 凋亡:线粒体途径,涉及 p53 和 Bcl-2 家族蛋白的表达;以及未折叠蛋白反应途径,涉及 GRP78 和 CHOP 的表达。