紫外线照射对大鼠晶状体上皮细胞Ca2＋-ATP酶表达及活性影响的实验研究  

Effects of ultraviolet irradiation on expression of Ca^(2+)-ATPase and their activities in rat lens epithelial cells

作　　者：王道光[1] 郭大东[2] 毕宏生[2] 杜然然[2] 吴秋欣[1] 刘冬梅[2] 

机构地区：[1]山东中医药大学第二临床学院,山东省济南市250014 [2]山东中医药大学眼科研究所、山东省高校中西医结合眼病防治技术重点实验室,山东省济南市250002

出　　处：《眼科新进展》2013年第11期1016-1020,共5页Recent Advances in Ophthalmology

基　　金：山东省自然科学基金资助(编号:ZR2010HM032)~~

摘　　要：目的研究紫外线对大鼠晶状体上皮细胞(lens epithelial cell,LEC)Ca2+-ATP酶表达及活性的影响。方法 6周龄的Wistar大鼠随机分为对照组和实验组,对照组大鼠未照射紫外线,实验组分别给予紫外线照射1 d、3 d和7 d,照射完毕后摘除双眼眼球,取出晶状体。利用实时荧光定量PCR检测紫外线对LEC Ca2+-ATP酶在mRNA水平表达的影响,Western blot检测紫外线对LEC Ca2+-ATP酶在蛋白水平表达的影响,化学比色法检测LEC内超微量Ca2+-ATP酶活性的变化,HE染色观察LEC的形态学改变,免疫组织化学SP法检测LEC中Ca2+-ATP酶的表达情况。结果紫外线照射1 d、3 d、7 d后,大鼠LEC中Ca2+-ATP酶mRNA、蛋白和活性表达水平逐渐下降,其中ATP2A2和ATP2C2的mRNA表达水平分别是对照组的(0.60±0.10)倍、(0.20±0.05)倍、(0.05±0.01)倍和(0.40±0.10)倍、(0.10±0.05)倍、(0.02±0.01)倍;Ca2+-ATP酶蛋白相对灰度值分别是对照组的(0.66±0.01)倍、(0.46±0.02)倍和(0.15±0.01)倍;Ca2+-ATP酶活性表达水平分别是对照组的0.87倍、0.61倍及0.22倍。以上各实验组与对照组相比,差异均有统计学意义(均为P<0.05)。实验组LEC细胞形态学亦发生明显变化,免疫组织化学检测大鼠LEC Ca2+-ATP酶表达逐渐下降,平均光密度值与对照组比较差异均有统计学意义(均为P<0.05)。结论紫外线照射可降低大鼠LEC Ca2+-ATP酶的表达,且呈时间和剂量依赖性,提示Ca2+-ATP酶对紫外线诱导的LEC凋亡、维持晶状体透明以及在白内障的发生发展中具有重要意义。Objective To investigate the effects of ultraviolet irradiation on the expression of Ca2＋-ATPase and their activities in rat lens epithelial cells （ LEC ）. Methods The 6-week-old Wistar rats were randomly divided into control group and experimental groups,no ultraviolet irradiation was given in control group ,and ultraviolet irradiation for 1 day,3 days and 7 days were given in experimental group. Further,the eyeballs of rats were extirpated and lenses were further extracted. The effects of ultraviolet on Ca2＋-ATPase expression were tested both by real-time quantitative PCR at mRNA level and by Western blot at the protein level in rat lens epithelial cells, the activity changes of Ca2＋-ATPase in lens epithelial cells were detected by colorimetric method, the morphological changes in rat LEC were investigated by HE staining. In the mean time,the expression of Ca2＋ -ATPase was analyzed by SP immunohistochemistry. Re- suits After exposure to ultraviolet irradiation for 1 day, 3 days and 7 days, it was found that Ca2＋-ATPase was decreased at mRNA, protein and activity levels. Compared with the expression level of relevant control, the mRNA levels for ATP2A2 and ATP2C2 in experimental group were （0.50 ± 0.10 ） times, （0.20 ±0.05 ） times, （0.05 ± 0.01 ） times and（0.40 ± 0.10） times, （ 0. 10 ± 0.05 ） times and （ 0.02 ± 0.01 ） times, respectively. The total Ca2＋-ATPase protein level in experimental group was（0.66 ± 0.01 ）times, （0.46 ± 0.02）times and （0. 15 ± 0.01 ）times compared with control group. In the meantime, Ca2＋-ATPase activity was 0.87 times,0.61 times and 0.22 times compared with the control group. There were statistical differences in above indexes between two groups （ all P 〈 0.05 ）. The morphological changes markedly occurred in LEC and the expression of Ca2＋-ATPase was gradually decreased by means of immunohistochemistry. Conclusion Ultraviolet irradiation can reduce the expression of Ca2＋-ATPase In a dose-dependent manner in rat LEC,

关 键 词：紫外线 大鼠 晶状体上皮细胞 CA2+-ATP酶 细胞凋亡 

分 类 号：R-332[医药卫生]