丙戊酸钠对大鼠视网膜缺血再灌注所致视网膜损伤的保护作用  

Protective effect of valproic acid on ischemia-reperfusion induced injury in retina of rat

作　　者：张珍珍[1] 吴星伟[1] 宫媛媛[1] 张薇[1] 尹莉莉[1] 

机构地区：[1]上海交通大学医学院附属上海市第一人民医院眼科,200080

出　　处：《中华眼科杂志》2012年第8期739-743,共5页Chinese Journal of Ophthalmology

基　　金：上海市卫生局中医药重大课题基金（2006ZD01）

摘　　要：目的探讨丙戊酸钠对大鼠视网膜缺血再灌注所致视网膜损伤的保护作用及其相关机制。方法实验研究。Wistar大鼠90只，随机分为3组：空白对照组、视网膜缺血再灌注＋磷酸盐缓冲液组（实验对照组）和缺血再灌注＋丙戊酸钠组（实验组），每组30只。前房加压灌注方法制作大鼠视网膜缺血再灌注模型，于损伤后6、12、24、48h和7d分别取眼球做视网膜组织病理学检查，荧光金逆行标记后行视网膜神经节细胞计数；免疫印迹法检测视网膜组织中热休克蛋白（Hsp）70表达水平及转录因子Spl的乙酰化水平；逆转录（RT）一PCR法检测大鼠视网膜组织中bcl一2和bax的mRNA水平。采用配对t检验对所得数据进行统计学分析。结果（1）缺血后24h，视网膜组织HE染色示实验组视网膜水肿程度较实验对照组轻，差异有统计学意义（t=7．491，P〈0．05）；荧光金逆行标记示再灌注后7d，实验组存活的神经节细胞数[（1629±63）个／mm2]较实验对照组[（9084-65）个／mm2]明显增多，差异有统计学意义（t=7．248，P〈0．05）。（2）免疫印迹法检测结果显示，再灌注损伤后24h实验组视网膜中Hsp70蛋白表达水平及Spl的乙酰化水平升高，与实验对照组相比差异有统计学意义（t=6．176，11．264，P值均〈0．05）；RT—PCR法检测结果显示，再灌注损伤后24h实验组视网膜组织中bcl-2mRNA表达水平（0．403±0．009）较实验对照组（0．314±0．012）升高，差异有统计学意义（t=5．489，P〈0．05）；实验组baxmRNA表达水平（0．383±0．009）较实验对照组（0．492±0．016）降低，差异有统计学意义（t=5．723，P〈0．05）。结论丙戊酸钠对大鼠视网膜缺血再灌注损伤具有保护作用，Hsp70、bcl-2表达水平的上调及bax表达水平的下调可能与该作用相关。Objective To study the protective effects and the related mechanism of valproic acid （VPA） on ischemia-reperfusion-induced injury in retina of rat. Methods Experimental study. Ninety Wistar rats were divided randomly into three groups： normal （ blank ） control group, retinal ischemia-reperfusion （experimental control） group treated with PBS, retinal isehemia-reperfusion （experimental） group treated with VPA. Retinal ischemia was induced by acute high intraocular pressure. Rats were executed at 6,12,24, 48 h and 7 d after reperfusion. The eyeballs were enucleated for retinal histopathological examination. The fluoro-gold retrograde labeling was performed and the survival of retinal ganglion cells was analyzed by calculating the densities in fluoro-gold labeled retinal ganglion cell. The protein expression of heat shock protein 70（Hsp70） and the acetylation of transcription factor Spl were analyzed by Western blot assay. The levels of bcl-2 and bax mRNA were analyzed by RT-PCR assay. To determine the significance of differences, analysis of paired-samples t-test was cmTied out. Results （ 1 ） The HE staining of retinal histological section showed that the edema of retina were attenuated significantly by VPA in experimental group, the difference between the experimental group and the experimental control group was statistically significant （ t = 7. 491,P 〈 0.05 ）. The fluoro-gold retrograde labeling showed that the survival of retinal ganglion cells in experimental group[ （ 1629± 63 ）/mm2 ] was significantly higher than experimental group [ （ 908 ± 65 ）/ mm2 ], the difference between the two groups was statistically significant （ t = 7. 248, P 〈 0. 05 ）. （ 2 ） The immuno-blot analysis showed that VPA resulted in significant increase in expression of Hsp70 protein in ischemic retinas,the difference between experimental group and experimental control group was statistically significant （t = 6. 176, P 〈 0. 05 ）. The acetylation of Spl was significantly higher in

关 键 词：丙戊酸 HSP70热休克蛋白质类 再灌注损伤 视网膜疾病 大鼠 

分 类 号：R965[医药卫生—药理学]