热休克预处理对人晶状体上皮细胞HSP60mRNA表达的影响及意义

发布时间：2018-02-12 16:04

本文关键词： 晶状体上皮细胞热休克蛋白60 热休克预处理 SYBR GreenⅠ 实时荧光定量PCR　出处：《南昌大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的采用SYBR Green I实时荧光定量PCR方法检测人晶状体上皮细胞在不同环境刺激处理后HSP60mRNA的表达的变化,探讨HSP60mRNA表达变化规律以及不同环境刺激处理后表达是否有差异。方法采用正常人组织源性晶状体上皮细胞HUM-CELL-0098进行细胞传代培养后,将传代培养后的细胞进行分组实验,在氧化条件以及热休克处理后加氧化条件下培养细胞30min后,在正常培养条件下继续培养并且选取不同时间点(Oh、2h、4h、6h、16h、24h)的细胞采用SYBR Green I实时荧光定量PCR检测HSP60mRNA的表达情况。结果观察到晶状体上皮细胞在不同环境刺激下均表达HSP60mRNA,但热休克预处理后的细胞HSP60mRNA的表达较未热休克处理细胞高。氧化应激条件下HSP60mRNA的表达明显增加,在刺激结束后表达含量为最高,以后逐渐下降；在热休克预处理组中HSP60mRNA的表达含量也明显增加,但在4h表达量最高,以后逐渐下降。两组细胞在不同时段HSP60mRNA表达量变化趋势不同F=178.786,P=0.007,P0.05,有统计学意义。此外,热休克预处理组与单纯氧化应激组中HSP60mRNA表达含量有显著差异,热休克处理组的表达含量较单纯氧化应激组高,存在组间差别F=193.118,P=0.001,P0.05,有统计学意义。时间与分表交互作用F=124.939,P=0.003,有统计学意义。两组细胞HSP60mRNA表达量进行各个时间两两对比发现这两组数据在统计学上有差异。此外,还发现热休克处理后的细胞表达HSP60mRNA可能具有时间依赖性。结论人晶状体上皮细胞在不同应激条件下,均能诱导HSP60mRNA的合成,但在不同应激条件下的HSP60mRNA的表达是具有差异的,热休克预处理后的细胞HSP60mRNA的表达高于氧化损伤处理的细胞,给予预先的热休克处理可通过提高HSP60mRNA表达来保护晶状体。
[Abstract]:Purpose. SYBR Green I real-time quantitative PCR was used to detect the changes of HSP60mRNA expression in human lens epithelial cells treated with different environmental stimuli, and to investigate the changes of HSP60mRNA expression and whether there were differences in the expression of HSP60mRNA after different environmental stimuli. Method. Normal human tissue-derived lens epithelial cells (HUM-CELL-0098) were cultured by subculture. The cultured cells were divided into two groups. The cells were cultured for 30 minutes under the conditions of oxidation and heat shock. SYBR Green I real-time fluorescence quantitative PCR was used to detect the expression of HSP60mRNA. Results. It was observed that HSP60mRNAs were expressed in lens epithelial cells under different environmental stimuli, but the expression of HSP60mRNA was higher in HSP60mRNAs treated with heat shock than that in cells without heat shock. The expression of HSP60mRNAs was significantly increased under oxidative stress. The expression level of HSP60mRNA increased significantly in heat shock preconditioning group, but the highest expression level was observed at 4 h after heat shock preconditioning. The change trend of HSP60mRNA expression in the two groups was different at different time, there was significant difference in the expression of HSP60mRNA between heat shock preconditioning group and oxidative stress group, and there was significant difference between heat shock preconditioning group and oxidative stress group. The expression level in heat shock treatment group was higher than that in oxidative stress group. There was a significant difference between the two groups (P 0.05). The interaction between time and time was statistically significant. The expression of HSP60mRNA in the two groups was compared at different times. In addition, there was statistical difference between the two groups. It was also found that the expression of HSP60mRNA in the cells treated with heat shock may be time dependent. Conclusion. Human lens epithelial cells could induce the synthesis of HSP60mRNA under different stress conditions, but the expression of HSP60mRNA was different under different stress conditions. The expression of HSP60mRNA in cells pretreated with heat shock was higher than that in cells treated with oxidative damage. Preconditioning with heat shock can protect the lens by increasing the expression of HSP60mRNA.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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