复方水蛭滴眼液抑制H_2O_2诱导的晶状体上皮细胞凋亡的信号转导机制研究

The Signal Transduction Mechanism of Inhibitory Effect of Compound Leech Eye Drops on Apoptosis of Lens Epithelial Cells

目的探讨复方水蛭滴眼液抑制H 2O2诱导的晶状体上皮细胞(LEC)凋亡的信号转导机制。方法采用牛LEC进行原代和传代培养,用H 2O2诱导LEC凋亡,同时加入终浓度为1/1000的复方水蛭滴眼液共同孵育。荧光分光光度法及放射免疫法检测LEC内游离钙([Ca2+]i)、cAM P和cGM P浓度变化。结果H 2O2可明显升高LEC内[Ca2+]i和cAM P的水平,显著降低cGM P水平;加入复方水蛭滴眼液后,细胞内[Ca2+]i和cAM P浓度较H 2O2组显著降低,而cGM P水平显著升高。结论复方水蛭滴眼液抑制H 2O2诱导的LEC凋亡的机制可能是通过降低LEC内[Ca2+]i和cAM P的水平,阻断或抑制Ca2+-钙调蛋白依赖性蛋白激酶途径、Ca2+-蛋白激酶C途径以及cAM P-蛋白激酶A(PKA)信号转导途径抑制LEC凋亡;同时通过升高LEC内cGM P浓度、激活cGM P-蛋白激酶C(PKC)信号转导途径抑制LEC凋亡。所以,复方水蛭滴眼液是通过多条途径抑制H 2O2诱导的LEC凋亡。

Objective： To investigate the mechanism of signal transduction of compound leech eye drops（Co - leech） on inhibiting apoptosis of lens epithelial cells（LEC） induced by H2O2. Methods： Bovine LEC were primarily cultured and subcultured. The LEC were incubated with H2O2 to induce LEC apoptosis. The LEC damaged by oxidative stress of H2O2 were incubated with Co - leech （1/1000）. Then the [ Ca^2 ＋ ] i, cyclic adenosine monophosphate （cAMP） and cyclic gnanosine monophosphate（cGMP） concentration of cultured LEC were examined. Results： Co-leech decreased the concentration of [Ca^2 ＋]i and cAMP of LEC, but increase the concentration of cGMP. Conclusion： The mechanism of signal transduction of compound Co - leech on inhibit apoptosis of LEC maybe related to the cross- talk among calcium signal transduction system, cAMP signal transduction system and cGMP signal transduction system.