Role of glutathione and other antioxidants in the inhibition of apoptosis and mesenchymal transition in rabbit lens epithelial cells.
Role of glutathione and other antioxidants in the inhibition of apoptosis and mesenchymal transition in rabbit lens epithelial cells.
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DOI：
10.4238/2014.september.1.1
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发表时间：
2014-09
期刊：
影响因子：
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通讯作者：
H. J. Wang;J. Zhu;G. Zheng
中科院分区：
文献类型：
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作者：
H. J. Wang;J. Zhu;G. Zheng
本研究旨在探讨谷胱甘肽（GSH）和过氧化氢酶（CAT）在转化生长因子β（TGF-β）诱导的透镜上皮细胞（LEC）凋亡和上皮间质转化（EMT）中的作用及机制，以防治白内障。随机选取4周龄健康家兔，取其晶状体进行体外培养。将第二代和第三代细胞分为6组（A组：75 pg/mL TGF-β2; B组：75 pg/mL TGF-β2+10 mM GSH; C组：75 pg/mL TGF-β2+300 U/mL CAT; D组：10 mM GSH; E组：300 U/mL CAT; F组：对照组）。倒置显微镜下观察细胞形态。加入75 pg/mL TGF-β2后，细胞差距增大，细胞呈网状，细胞肿胀，呈梭形。但是，抗氧化剂降低了这些变化。在12、24和48 h时分析生长抑制，组间差异无统计学显著性。细胞凋亡分析，A组与B、C组比较差异有统计学意义（P<0.05）。逆转录-聚合酶链反应检测α-SMA mRNA的表达。A组α-SMA mRNA水平明显高于B、C组（P<0.05）。TGF-β2抑制LEC增殖，诱导细胞凋亡和EMT。GSH和CAT抑制LECs的凋亡和EMT，对细胞增殖影响不大。TGF-β2作为一种细胞信号分子，可能参与了细胞凋亡和EMT的发生。
This study aimed to discuss the effect and mechanism of glutathione (GSH) and catalase (CAT) on transforming growth factor beta (TGF-β)-induced lens epithelial cell (LEC) apoptosis and epithelial mesenchymal transition (EMT) to prevent and control cataracts. Healthy rabbits (4 weeks old) were randomly selected, and LECs from their lenses were cultured in vitro. The 2nd- and 3rd-generation cells were divided into 6 groups (group A: 75 pg/mL TGF-β2; B: 75 pg/mL TGF-β2+10 mM GSH; C: 75 pg/mL TGF-β2+300 U/mL CAT; D: 10 mM GSH; E: 300 U/mL CAT; and F: control group). Cell morphology was observed under an inverted microscope. The gap between cells increased, and the cells became reticulate after adding 75 pg/mL TGF-β2; also, the cells swelled and appeared spindle-shaped. However, antioxidants reduced these changes. Growth inhibition was analyzed at 12, 24, and 48 h, and the differences between groups were not statistically significant. Cell apoptosis was analyzed, and the differences between group A and groups B and C were statistically significant (P<0.05). Reverse transcriptase-polymerase chain reaction was used to detect mRNA expression of α-SMA. The α-SMA mRNA level was greater in group A than in groups B and C (P<0.05). TGF-β2 inhibited LEC proliferation and induced apoptosis and EMT. GSH and CAT inhibited apoptosis and EMT in LECs, and they had little effect on cell proliferation. Reactive oxygen species may be involved in cell apoptosis and EMT induced by TGF-β2 as a cell-signaling molecule.