珍珠明目滴眼液改善TNF-α和IFN-γ诱导人角膜上皮细胞损伤的作用  

Effects of Zhenzhu Mingmu Eye Drops on improving the human corneal epithelial cells injury induced by TNF-α and IFN-γ

作　　者：吴慧敏 赛那 杜雪莹[1] 尹兴斌[1] 董晓旭[1] 黄平情 倪健[1] 曲昌海[1] WU Hui-min;SAI Na;DU Xue-ying;YIN Xing-bin;DONG Xiao-xu;HUANG Ping-qing;NI Jian;QU Chang-hai(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;College of Pharmacy,Inner Mongolia Medical University,Hohhot 010110,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]内蒙古医科大学药学院,呼和浩特010110

出　　处：《中华中医药杂志》2022年第4期2311-2315,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：北京市自然科学基金项目(No.7202123)。

摘　　要：目的:探讨珍珠明目滴眼液(ZMED)对肿瘤坏死因子α(TNF-α)和干扰素γ(IFN-γ)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150 ng/mL的TNF-α和IFN-γ(1∶1)溶液刺激离体培养的HCEC建立干眼损伤模型,药物组分别给予稀释倍数为1000、100、50倍的ZMED;分别采用MTT实验检测ZMED对TNF-α和IFN-γ诱导损伤HCEC的保护作用;采用AO/EB双染法和Hoechst荧光染色法观察各组细胞凋亡形态的差异;采用流式细胞术分析各组细胞凋亡率及细胞周期分布;通过Western Blot检测凋亡相关蛋白表达水平的变化。结果:ZMED稀释50倍即可以明显增加TNF-α和IFN-γ诱导损伤的HCEC的存活率;ZMED稀释100倍以下可呈浓度依赖性地降低HCEC凋亡率并抑制其G2期阻滞(P<0.05);ZMED给药后cleaved Caspase-8、cleaved Caspase-3、PARP、p-ERK1/2/ERK1/2下调(P<0.05),Bcl-2/Bax上调(P<0.05)。结论:ZMED对150 ng/mL的TNF-α和IFN-γ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Caspase-8/Caspase-3信号通路,Bcl-2家族蛋白与ERK1/2的活化相关。Objective:To investigate the protective effect and mechanism of Zhenzhu Mingmu Eye Drops(ZMED)on human corneal epithelial cells(HCEC)injury induced by TNF-αand IFN-γ.Methods:The dry eye model was established by stimulating HCEC cultured in vitro with 150 ng/mL TNF-αand IFN-γ(1:1)solution,and 1000,100,50 different dilution rates of ZMED were given to the treatment groups.MTT assay was used to evaluate the protective effect of ZMED on HCEC induced by TNF-αand IFN-γ.AO/EB double staining and Hoechst fluorescence staining were used to observe the morphological differences of apoptosis.The apoptosis rate and cell cycle distribution of each group were analyzed by flow cytometry.Moreover,the expression levels of apoptosis-related proteins were detected by Western Blot.Results:A total of 50 fold dilution of ZMED significantly increased the survival rate of HCEC injured by TNF-αand IFN-γ.ZMED diluted less than 100 times could reduce the apoptosis rate and inhibit phase arrest of G2 in a dose-dependent manner(P<0.05).Furthermore,after ZMED administration,the levels of cleaved Caspase-8,cleaved Caspase-3,cleaved PARP and p-ERK/ERK1/2 ratio were down-regulated(P<0.05),the Bcl-2/Bax rate was up-regulated(P<0.05).Conclusion:ZMED has a significant protective effect on HCEC induced by 150 ng/mL TNF-αand IFN-γsolution,which might be related to the inhibition of Caspase-8/Caspase-3 signaling pathway and the activation of Bcl-2 family proteins and ERK1/2.

关 键 词：珍珠明目滴眼液 干眼 人角膜上皮细胞 肿瘤坏死因子α 干扰素Γ 凋亡相关蛋白 

分 类 号：R285.5[医药卫生—中药学]