紫外线照射对SD大鼠晶状体上皮细胞p53、Bax、Bcl-2表达的影响

发布时间：2018-06-22 02:48

本文选题：紫外线 + 晶状体上皮细胞　；参考：《眼科新进展》2017年05期

【摘要】：目的探讨p53、Bax、Bcl-2基因在紫外线诱导晶状体上皮细胞(lens epithelial cell,LEC)凋亡中的作用。方法 6周龄SD大鼠40只随机分为对照组和实验组,对照组不做处理,实验组腹腔注射100 g·L-1水合氯醛(0.35 mL/100 g),以波长为300~320 nm的紫外线照射眼球15 min,分别于紫外线照射后1、3、5、7 d处死并取出晶状体。Hoechst33258染色法观察LEC的形态变化,实时定量PCR及免疫组织化学检测p53、Bax、Bcl-2mRNA和蛋白在LEC中表达水平。结果紫外线照射后晶状体发生混浊,其LEC发生凋亡,细胞排列不规则、大小不一。实时定量PCR检测结果显示,与对照组相比,紫外线照射后1 d Bax mRNA表达水平升高(P0.05),照射后3、5、7 d Bax和p53 mRNA表达水平均升高(均为P0.01);与对照组相比,Bcl-2 mRNA表达水平在照射后1 d降低(P0.01),而照射后5 d和7 d则升高(均为P0.01)。紫外线照射后p53与Bax mRNA相对表达水平的变化呈正相关(r=0.952,P0.01)。免疫组织化学结果显示,紫外线照射后LEC中p53和Bax蛋白表达逐渐增强,且由细胞质向细胞核转移;相反照射后1 d Bcl-2蛋白表达降低,而3、5、7 d则逐渐增高。结论紫外线诱导LEC凋亡与p53介导Bax/Bcl-2的表达调控有关。
[Abstract]:Objective to investigate the role of p53 Baxfen Bcl-2 gene in ultraviolet induced apoptosis of lens epithelial cells (lens epithelial cells). Methods Forty 6-week-old SD rats were randomly divided into two groups: control group and experimental group. In the experimental group, 100g L-1 chloral hydrate (0.35mL / 100 g),) was injected intraperitoneally into the eyeball for 15 mins with ultraviolet radiation wavelength of 300-320 nm. The rats were killed on the 7th day after ultraviolet irradiation, and the morphologic changes of LEC were observed by Hoechst33258 staining. The expression of Bcl-2 mRNA and protein in LEC was detected by real-time quantitative PCR and immunohistochemistry. Results after ultraviolet irradiation, lens opacity, LEC apoptosis and irregular cell arrangement were observed. The results of real-time quantitative PCR showed that, compared with the control group, The expression of Bax mRNA increased 1 day after ultraviolet irradiation (P0.05), and the expression of Bax and p53 mRNA increased on the 7th day after irradiation (both P0.01), and the expression level of Bcl-2 mRNA decreased at 1 day after irradiation (P0.01), but increased on the 5th and 7th day after irradiation (P0.01). There was a positive correlation between p53 and Bax mRNA expression after UV irradiation (r = 0.952, P 0.01). Immunohistochemical results showed that the expression of p53 and Bax protein in LEC increased gradually after ultraviolet irradiation and transferred from the cytoplasm to the nucleus, whereas the expression of Bcl-2 protein decreased at 1 day after irradiation, but increased gradually on the 7th day after irradiation. Conclusion UV induced apoptosis of LEC is related to p53 mediated Bax-Bcl-2 expression.
【作者单位】：遵义医学院生物化学与分子生物学教研室;
【基金】：贵州省优秀科技教育人才省长专项基金资助项目(2009047)~~
【分类号】：R776.1

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