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  

Effects of ultraviolet radiation on p53,Bax and Bcl-2 expression in rat lens epithelial cells

作　　者：赵学发[1] 杨名慧[1] 葛正龙[1] 

机构地区：[1]遵义医学院生物化学与分子生物学教研室,贵州省遵义市563099

出　　处：《眼科新进展》2017年第5期423-427,共5页Recent Advances in Ophthalmology

基　　金：贵州省优秀科技教育人才省长专项基金资助项目(2009047)~~

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(P<0.05),照射后3、5、7 d Bax和p53 mRNA表达水平均升高(均为P<0.01);与对照组相比,Bcl-2 mRNA表达水平在照射后1 d降低(P<0.01),而照射后5 d和7 d则升高(均为P<0.01)。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,P<0.01)。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。Objective To explore the roles of p53 ,Bax and Bcl-2 in rat lens epithelial ceils apoptosis induced by ultraviolet radiation. Methods Healthy Sprague- Dawley rats （40,6-weeks old,150 g） were selected and divided into 5 groups randomly. After SD rats were injected 100 g · L^-1 chloral hydrate （0.35 mL/100 g） intraperitoneally and pupil dilation,the eyes of SD rats were radiated 15 minutes by using UV-B （300 - 320 run）. The exposed animals were sacrificed at 1 day,3 days,5 days and 7 days after exposure. Both lenses from all animals were extracted. The apoptosis of rats lens epithelial cells was performed by Hoechst 33258 staining. The expression of p53, Bax and Bcl-2 in rat lens epithelial cells at mRNA level was detected by real-time PCR. The distribution and expression of p53, Bax and Bcl-2 were observed by immunohlstochemistry. nesults The apoptosis of lens epithelial cells was aggravated after exposure. The expression of p53 and Bax at mRNA and protein level was increased after UV exposure （P 〈 0.01 ）. But for Bcl-2 the expression both at mRNA and protein level was decreased after a 1 day-exposure（P 〈 0.01 ） and was increased after UV exposure of 5 days and 7 days（P 〈 0.01 ）. The mRNA expression level of p53 was positively correlated with the Bax （r=0. 952,P〈0.01 ）. The expression of p53 and Bax protein were increased after UV exposure, and transferred from cytoplasm to nucleus;While the expression of Bcl-2 protein was decreased at 1 day after UV exposure, and increased gradually at 3 days, 5 days and 7 days. Conelusion The apoptotic process of rat lens epithelial cells induced by ultraviolet may be related with p53 regulating the expression of Bax/Bcl-2.

关 键 词：紫外线 晶状体上皮细胞 P53 BAX Bcl-2 

分 类 号：R776.1[医药卫生—眼科]