珍珠明目滴眼液改善高渗诱导角膜上皮细胞损伤的作用机制研究  

Study on effect and mechanism of Zhenzhu Mingmu Eye Drops protected HCEC against hypermeability-induced injury

作　　者：杜雪莹[1] 郑姣[1] 顾小盼 曲昌海[1] 倪健[1] 

机构地区：[1]北京中医药大学中药学院,北京100029

出　　处：《中草药》2017年第16期3425-3430,共6页Chinese Traditional and Herbal Drugs

摘　　要：目的探讨珍珠明目滴眼液(ZMED)对高渗诱导人角膜上皮细胞(HCEC)损伤的保护作用及相关机制。方法采用110 mmol/L氯化钠高渗溶液刺激离体培养的HCEC建立干眼损伤模型,药物组分别给予不同稀释倍数的ZMED;分别采用MTT法、乳酸脱氢酶(LDH)实验检测ZMED对高渗诱导损伤HCEC的保护作用;采用流式细胞术分析各组细胞凋亡率;采用AO/EB双染法和Hochest荧光染色法观察各组细胞凋亡形态的差异;通过Western blotting法检测Caspase凋亡通路上关键蛋白表达水平的变化。结果 ZMED可以明显增加高渗诱导损伤的HCEC存活率并降低其LDH释放量;ZMED稀释25倍以下作用于高渗诱导HCEC后,细胞凋亡率较模型组明显降低,且凋亡的HCEC数量呈浓度依赖性减少;ZMED给药后剪切态的Caspase-9、Caspase-3、PARP呈浓度依赖性下调。结论 ZMED对110 mmol/L氯化钠高渗溶液诱导损伤的HCEC有明显的保护作用,该作用可能与其抑制线粒体介导的Caspase-9/Casepase-3细胞调亡通路相关。Objective To investigate the effects and mechanism of Zhenzhu Mingmu Eye Drops（ZMED） on human corneal epithelial cells（HCEC） injury model induced by high permeability. Methods The dye eyes disease model was established using 110 mmol/L Na Cl to stimulate HCEC, and different concentration of ZMED was given to the treatment group. MTT assay and lactate dehydrogenase（LDH） leakage were used to evaluate the protection effects of ZMED on HCEC. The effects of ZMED on apoptosis of HCEC were analyzed by flow cytometry. AO/EB double staining and Hoechst 33258 analysis were used as sensitive assays for apoptosis. Moreover, HCEC treated with ZMED were subjected to Western blotting for protein levels of Caspase-9, Caspase-3 and PARP. Results ZMED significantly increased cell viability of HCEC and decreased the LDH release in a dose-dependent manner. ZMED dramatically decreased the apoptosis rate. Furthermore, ZMED down-regulated the levels of cleaved Caspase-9, cleaved Caspase-3 and cleaved PARP. Conclusion ZMED could protect HCEC against hypermeability, which may be related to the inhibition of Caspase-9/Casepase-3 signaling pathways.

关 键 词：珍珠明目滴眼液 干眼 人角膜上皮细胞 高渗 凋亡 乳酸脱氢酶 CASPASE-9 CASEPASE-3 

分 类 号：R285.5[医药卫生—中药学]