摘要
目的研究急性高眼压大鼠视网膜的内质网应激反应(endoplasmic reticulum stress,ERS)以及活血化瘀汤对ERS的影响。方法 88只Wistar大鼠随机分为正常对照组(8只)、高眼压组(40只)、治疗组(40只)。除正常对照组外所有大鼠均随机取一眼制作急性高眼压模型,治疗组在造模后给予活血化瘀汤(4mL·d-1)灌胃,高眼压组和治疗组分别在造模后12h、1d、3d、7d、14d均等处死大鼠。HE染色观察各组视网膜组织结构变化,免疫组织化学染色观察视网膜GRP78、Thy-1表达变化,RT-PCR检测视网膜GRP78、Thy-1 mRNA变化。结果光镜下见高眼压组视网膜早期水肿,后期萎缩;治疗组与高眼压组在各时间点的变化趋势相同,但程度均轻于高眼压组。免疫组织化学和RT-PCR结果显示高眼压组造模后12h,GRP78蛋白和mRNA的表达量迅速上调(0.291±0.012、1.534±0.143),与正常对照组(0.195±0.003、1.011±0.011)比较,差异均有统计学意义(均为P<0.05);造模后1d,表达量达到高峰(0.498±0.049、2.047±0.177);造模后3d迅速降低,但仍高于正常对照组,差异均有统计学意义(均为P<0.05)。治疗组GRP78蛋白和mRNA表达量在造模后12h(0.274±0.023、1.398±0.031)较正常对照组显著增多,差异均有统计学意义(均为P<0.05),较高眼压组低,但差异均无统计学意义(均为P>0.05);造模后1d达高峰(0.432±0.042、1.641±0.164),但增加量均少于同时间点高眼压组,差异均有统计学意义(均为P<0.05);造模后3d表达量降低(0.212±0.022、1.174±0.126),与正常对照组比较,差异均无统计学意义(均为P>0.05),但低于同时间点高眼压组,差异均有统计学意义(均为P<0.05)。造模后7d、14d三组间GRP78蛋白和mRNA的表达量差异均无统计学意义(均为P>0.05)。造模后14d,高眼压组和治疗组Thy-1蛋白和mRNA的表达量均降低,但治疗组Thy-1 mRNA表达量(0.754±0.053)明显高于高眼压组(0.627±0.069),差异有统计学意义(P<0.05)。结论ERS参与了急性高眼压视网膜损伤过程,中药活血化瘀汤对该损伤有保护作用,可能与其抑制ERS有关。
Objective To study endoplasmic reticulum stress （ERS） in retina of rats with acute intraocular hypertension and investigate effects of Huoxuehuayu decoction on it. Methods A total of 88 Wistar rats were randomly divided into normal control group （ 8 rats）, intraocular hypertension group （ 40 rats ） and treatment group （ 40 rats）. One eye of each rat in intraocniar hypertension group and treatment group was ran- domly selected to establish the model of acute intraocular hypertension. Huoxuehuayu decoction gavage was administered to rats in treatment group after modeling. Rats in intraocular hypertension group and treatment group were evenly executed at five time-points： 12 hours, 1 day,3 days,7 days, 14 days after modeling. Structural changes in retinal issues were observed by Hematoxylin and eosin （HE） staining. Changes of expression levels of GRP78 and Thy-1 were detected by immunohistochemistry staining. Changes of expression levels of GRP78 mRNA and Thy-1 mRNA were detected by RT-PCR. Results Retinal edema was observed in intraocular hypertension group under microscopy at early stage and retinal atrophy at later stage. Treatment group shared tendencies of condition changes of intraocular hypertension group, but had milder conditions. Immunohistochemistry staining and RT-PCR results showed that expression levels of GRP78 protein and GRP78 mRNA of intraocular hypertension group gained a rapid rise in 12 hours after modeling （ 0. 291 ± 0.002,1. 534 ± 0.413 ）, compared with those of normal control group （0. 195 ± 0. 003.1.011 ± 0.011 ） （ both P 〈 0.05 ）. The exnression levels neaked at the2nd time-in-point （0.498 ± 0. 049 , 2. 047 ± 0. 177 ）. At the 3rd time-in-point, the expression levels decreased greatly, but were still significantly higher than those of normal control group （ both P 〈 0.05 ）. Expression levels of GRP78 protein and GRP78 mRNA in treatment group at the 1 st time-in-point （ 0.274 ± 0. 023,1. 398 ± 0.031 ） were significantly higher than those of norreal control group （ both P 〈 0.05 ）, but were lower than those of intraoeular hypertension group （ both P 〉 0.05 ）. The expression levels peaked at the 2nd time-in-point （0.432 ± 0.042,1. 641 ± 0. 164 ）, but the increments were lower than those of the intraocular hypertension group at the same time-in-point （ both P 〈 0.05 ）. At the ]rd time-in-point, the expression levels decreased （0.212 ± 0.022,1. 174 ± 0. 126 ）, which were not significantly different from those of the normal control group （ both P 〉 0.05 ）, but were significantly lower than those of the intraocular hypertension group at the same time-in-point （ both P 〈 0.05 ）. At the 4th and 5th time-in-points, differences among expression levels of GRP78 protein and GRP78 mRNA of the three groups were of no statistical significance （ all P 〉 0.05 ）. At the 4th time-in-point, expression levels of Thy-1 protein and Thy-1 mRNA of both intraocular hypertension group and treatment group decreased, but expression level of Thy-1 mRNA of treatment group（0. 754 ± 0. 053 ） was significantly higher than that of intraocular hypertension group （ 0. 627 ± 0. 069） （P 〈 0.05 ）. Conclusion ERS is a part of the process of retinal damage induced by acute ocular hypertension. Huoxuehuayu has a protective effect against the damage, which may be associated with its inhibitory effect on ERS.
出处
《眼科新进展》
CAS
北大核心
2013年第7期622-626,共5页
Recent Advances in Ophthalmology
关键词
内质网应激
活血化瘀汤
急性高眼压
葡萄糖调节蛋白78
endoplasmic reticulum stress
Huoxuehuayu decoction
acute intraoc-i ular hypertension
glucose-regulated protein 78
