摘要
目的观察活血化瘀方剂对大鼠视网膜急性高眼压损伤的保护作用及对低氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)表达的影响。方法 140只清洁级Wistar大鼠随机分为对照组10只、穿刺组10只、加压组60只和治疗组60只。前房灌注加压法制作急性高眼压模型维持60min(眼压至66mmHg,1kPa=7.5mmHg)。加压组分别于造模后2h、12h、1d、3d、7d、14d处死动物,治疗组在造模后即刻给予活血化瘀方剂4mL灌胃,并分别在造模后2h、12h、1d、3d、7d、14d处死动物。通过HE染色观察视网膜的形态学变化,免疫组织化学方法和实时定量逆转录聚合酶链反应(Real-TimeqRT-PCR)方法检测HIF-1α在各组视网膜中的表达情况。结果造模加压后1d,光学显微镜下可见视网膜高度水肿,7d时水肿基本消失,14d时视网膜各层结构紊乱,明显萎缩变薄。治疗组造模加压后2h、12h视网膜形态学变化与加压组同一时间点无明显差别,其他各时间点视网膜损伤程度均较加压组同一时间点轻。免疫组织化学染色及Real-Time qRT-PCR结果显示:造模加压后2 h HIF-1α的表达即增加(IOD0.54±0.06,mRNA1.5162±0.1199),12h达到高峰(IOD1.73±0.10,mRNA2.9041±0.1690),随着时间推移表达逐渐减少,14d时降至最低(IOD0.52±0.03,mRNA1.2847±0.0434),但仍高于对照组(IOD0.25±0.04,mR-NA1.0009±0.0480,P<0.05);治疗组该因子的表达变化趋势与加压组相同,但除造模后2h、12h外,治疗组各时间点HIF-1α的表达量均比加压组同一时间点低,差异均有统计学意义(均为P<0.05)。结论活血化瘀方剂对急性高眼压造成的视网膜损伤有保护作用,这种保护作用可能与其减少视网膜内源性HIF-1α的表达有关。
Objective To observe the protective effects of Huoxuehuayu prescription against the retinal damage caused by acute intraocular hypertension in rats,and its influence on hypoxia-inducible factor-1 alpha （HIF-1α） expression. Methods One hundred and forty Wistar rats were randomly divided into control group （10 rats）,punctural group （10 rats）,model group （60 rats） and treated group （60 rats）.Acute intraocular hypertension model was created by anterior chamber perfusion for 60 minutes （IOP=66 mmHg,1 kPa=7.5 mmHg）.Rats in treated group were administered with Huoxuehuayu prescription 4 mL immediately after pressurizing.The rats in different groups were sacrificed respectively at 2 hours,12 hours,1 day,3 days,7 days,14 days after pressurizing.The histological change of retina was observed by HE staining under the light microscope.The expression of HIF-1α in retina was determined by immunohistochemistry and Real-time quantitative reverse transcription polymerase chain reaction （Real-Time qRT-PCR）. Results In the model group,histological examination showed that the edema of retina was obvious at 1 day after pressurizing;the edema was disappeared at 7 days,and the retinal structure was disarranged with obvious atrophy at 14 days.The extent of retinal injury in the treated group was less severe than the model group at the same time stages except for the 2 hours and 12 hours.Immunohistochemistry and Real-Time qRT-PCR showed that in model group the expression of HIF-1α increased at 2 hours（IOD 0.54±0.06,mRNA 1.516 2±0.119 9）after pressurizing,got to the peak at 12 hours（IOD 1.73±0.10,mRNA 2.904 1±0.169 0）and decreased gradually,reduced to the minimum at 14 days（IOD 0.52±0.03,mRNA 1.284 7±0.043 4）,but higher than control group（IOD 0.25±0.04,mRNA 1.000 9±0.048 0,P〈0.05）.The trend of HIF-1α in treated group was similar to model group,but the expression of HIF-1α in the former were less than that in the later at the same time stages except for the 2 hours and 12 hours （
出处
《眼科新进展》
CAS
北大核心
2012年第9期818-822,共5页
Recent Advances in Ophthalmology
关键词
活血化瘀方剂
低氧诱导因子-1Α
急性高眼压
视网膜
Huoxuehuayu prescription
hypoxia-inducible factor-1 alpha
acute intraocular hypertension
retina
