摘要
目的观察渗透压对体外培养的兔角膜上皮细胞的影响及牛磺酸对高渗环境细胞的保护作用,为人工泪液的处方制定提供理论依据。方法采用分离培养的兔角膜上皮细胞,分为低渗组(143,229和257 mOsm·L^-1)、等渗对照组(286 mOsm·L^-1)和高渗组(315,343和429 mOsm·L^-1),用不同渗透压的培养液作用3 h后,换回正常培养液培养3 d,记录作用3 h及恢复培养3 d时的细胞形态,并用MTT法检测作用3 h、恢复培养1 d和恢复培养3 d后的细胞活性。在正常培养液286 mOsm·L^-1中加入牛磺酸培养细胞24 h后,换为429 mOsm·L^-1的高渗培养液培养3 h后,换回含牛磺酸的等渗培养液恢复培养1 d,倒置显微镜下记录作用3 h及恢复培养1 d时的细胞形态,并用MTT法检测细胞活性。结果高渗环境会导致细胞活性降低,甚至死亡;低渗环境会改变细胞形态及活性。257 mOsm·L^-1渗透压组与等渗对照组相比,在作用3 h及恢复培养1和3 d后细胞形态及细胞活性差异无统计学意义。含有不同浓度牛磺酸培养基的各组细胞也有轻度肿胀,高渗培养基作用3 h后以及恢复正常培养基培养1 d后,细胞活性差异具有显著统计学意义(P<0.05)。结论泪液高渗透压会造成眼表损伤。应用渗透压为257 mOsm·L^-1的人工泪液,在对正常细胞无影响的条件下,可降低干眼患者泪液的高渗透压,有助于干眼病的治疗。牛磺酸作为细胞保护剂,可降低高渗透压对细胞的损害,在干眼病治疗方面,具有很好的应用前景。
Objective To observe the effect of osmolarity on rabbit corneal epithelial cells cultured in vitro,and the protective effect of taurine on cells in hypertonic environments,therefore to provide a theoretical basis for the choice of the formulation and selection of artificial tears.Methods The rabbit corneal epithelial cells were cultured in hypo-osmolarity group(143,229 and 257 mOsm·L^-1),iso-osmolarity control group(286 mOsm·L^-1)and hyper-osmolarity group(315,343 and 429 mOsm·L^-1)for 3 h.Then they were returned to normal culture for 3 d,and their morphology were recorded.MTT assay was used to detect the viability of the cells in each group after 3 h,1 d and 3 d in recovery culture.The cells were cultured in the normal cultural medium(286 mOsm·L^-1)with taurine for 24 h,then were transferred to the cultural medium with the hyper-osmolarity of 429 mOsm·L^-1 for 3 h,finally were recovered the normal cultural medium with taurine for 1 d.The cell morphology was recorded by the inverted microscope and the cell activity were detected by MTT assay.Results Hypertonic environment may lead to the reduction of cell activity or even death.Hypotonic environment would change the cell morphology and activity.Compared with the iso-osmolarity control group,the osmolarity of 257 mOsm·L^-1 group had no significant difference in cell morphology and cell viability after 3 h and 1 d and 3 d of restoration.Although the cells cultured in medium with different concentrations of taurine had mild swelling,after immersing in the hypertonic medium for 3 h and restoring in normal medium for 1 d,the cell activity was significantly different from that of the group without taurine(P<0.05).Conclusion The hyper-osmolarity of lacrimal fluid may lead to ocular surface damage.With no effect on normal cells,the application of the artificial lacrimal fluid with osmotic pressure of 257 mOsm·L^-1 can reduce the hypertonic pressure of tear in dry eyes and help to treat dry eyes.Taurine,as a cytoprotective agent,could reduce the damage of hyperosmotic pressure to cells,and has a good application prospect in the treatment of dry eyes.
作者
潘士印
肖湘华
赵娜
安娜
刘先宁
朱秀萍
银勇
王养正
PAN Shiyin;XIAO Xianghua;ZHAO Na;AN Na;LIU Xianning;ZHU Xiuping;YIN Yong;WANG Yangzheng(Department of Ophthalmology,Xi′an No.1 Hospital,Shaanxi Institute of Ophthalmology,Shaanxi Key Laboratory of Ophthalmology,Clinical Research Center for Ophthalmology Diseases of Shaanxi Province,the First Affiliated Hospital of Northwestern University,Xi′an 710002,China)
出处
《西北药学杂志》
CAS
2019年第4期509-513,共5页
Northwest Pharmaceutical Journal
基金
陕西省科技厅自然科学基金项目(编号:2010JM4011,2011JM4023,2013JM4039)
西安市卫生局科技计划项目(编号:J2012B02)
关键词
牛磺酸
渗透压
角膜上皮细胞
干眼
taurine
osmolarity
corneal epithelial cells
dry eye