[关键词]
[摘要]
目的:观察高糖培养的人晶状体上皮细胞在创伤刺激后Cyclin D1的表达情况。
方法:采用MTT法观察不同浓度葡萄糖对体外培养的人晶状体上皮细胞(HLEB3)作用24h后细胞增殖活性的影响,确定最适葡萄糖作用浓度。将HLEB3细胞分为高糖预处理组(高糖培养基预处理24h后再更换高糖培养基培养)和非高糖预处理组(正常培养基培养24h后再更换高糖培养基培养),根据更换高糖培养基时是否进行划伤处理将细胞分为对照组和划伤组,分别采用qRT-PCR和Western blot法检测各组细胞创伤后不同时间点Cyclin D1 mRNA和蛋白的表达情况。
结果:一定浓度范围的葡萄糖能够增强细胞增殖活性,25.5mmol/L葡萄糖处理时细胞活性最强。高糖预处理细胞Cyclin D1表达在一定时间范围内呈时间依赖性表达下调; 非高糖预处理细胞Cyclin D1表达呈不规律性,在12h和48h处表达上调; 划伤处理可在一定程度上刺激细胞Cyclin D1表达上调。
结论:葡萄糖对人晶状体上皮细胞活性及Cyclin D1表达的影响呈现不规律性,创伤处理可在一定程度上上调细胞Cyclin D1的表达。
[Key word]
[Abstract]
AIM:To observe the expression pattern of Cyclin D1 in human lens epithelial cells(HLECs)after traumatic stimulation in high-glucose culture
METHODS: The activity of HLECs was detected by MTT method after incubate with differernt concentration glucose for 24h in vitro to determine the optimal glucose concentration. qRT-PCR and Western blot were used to detect the high glucose pretreatment group and the non-high glucose pretreatment group. The expression of Cyclin D1 in HLECs at different time points after traumatic stimulation was detected.
RESULTS: The viability of HLECs were increased when treatment with low concentration glucose, but the concentration should not exceed 25.5mmol/L, or it will inhibit the activity of HLECs; The reasult of high glucose pretreatment group reveal that the expression of Cyclin D1 is down-regulated in a time-dependent manner within a certain time range. While the expression of Cyclin D1 was irregular in the non-pretreatment group, it was increased at the time point of 12h and 48h. The score treatment can up-regulate the expression of Cyclin D1 in HLECs in a certain degreen.
CONCLUSION: The effects of glucose on HLECs activity and Cyclin D1 experssion are irrugular. Trauma treatment can stimulate the expression of Cyclin D1 in HLECs to some extent.
[中图分类号]
[基金项目]
海南省国家级大学生创新创业训练计划(No.201811810021); 海南医学院校级大学生创新创业训练计划(No.HYCX2018021)