CAJ | 학술논문

目的探讨第六类中间丝蛋白nestin在高眼压大鼠视网膜神经胶质细胞中的表达情况及意义.方法 42只SD大鼠采用烧灼右眼涡静脉的方法制作高眼压模型为模型组,剪开左眼结膜作为假手术组.测量并记录眼压,计数术后不同时间点视网膜神经节细胞(RGCs)数.Western blot半定量分析术后视网膜内nestin蛋白含量.免疫电镜检测nestin在高眼压大鼠视网膜Müller细胞上的诱导表达.行nestin/谷氨酰胺合成酶(GS)或nestin/胶质纤维酸性蛋白(GFAP)免疫荧光双标,共焦显微镜观察nestin/GFAP在Müller细胞上的表达情况.结果术后各时间点模型组与假手术组眼压的差异有统计学意义(P＜0.05).术后1～3周模型组RGCs数量显著减少,与假手术组比较差异有统计学意义(P＜0.05).免疫荧光结果显示术后2h～3周,模型组nestin在Müller细胞上表达的荧光强度逐渐增强,与假手术组比较差异有统计学意义.Western blot半定量分析结果与免疫荧光染色结果一致.免疫电镜结果进一步证实眼压升高后Müller细胞上出现nestin的诱导表达.结论 Nestin在Müller细胞上的诱导表达是Müller细胞对眼压升高产生的一种反应,nestin的表达量与病程进展相一致.眼压升高时,nestin在Müller细胞上诱导表达,尤其在足板处的强烈表达,可能对RGCs具有保护作用.Nestin有望成为一种新的研究视网膜损伤的生物标记物.
목적 탐토제륙류중간사단백nestin재고안압대서시망막신경효질세포중적표체정황급의의.방법 42지SD대서채용소작우안와정맥적방법 제작고안압모형위모형조,전개좌안결막작위가수술조.측량병기록안압,계수술후불동시간점시망막신경절세포(RGCs)수.Western blot반정량분석술후시망막내nestin단백함량.면역전경검측nestin재고안압대서시망막Müller세포상적유도표체.행nestin/곡안선알합성매(GS)혹nestin/효질섬유산성단백(GFAP)면역형광쌍표,공초현미경관찰nestin/GFAP재Müller세포상적표체정황.결과 술후각시간점모형조여가수술조안압적차이유통계학의의(P＜0.05).술후1～3주모형조RGCs수량현저감소,여가수술조비교차이유통계학의의(P＜0.05).면역형광결과 현시술후2h～3주,모형조nestin재Müller세포상표체적형광강도축점증강,여가수술조비교차이유통계학의의.Western blot반정량분석결과 여면역형광염색결과 일치.면역전경결과 진일보증실안압승고후Müller세포상출현nestin적유도표체.결론 Nestin재Müller세포상적유도표체시Müller세포대안압승고산생적일충반응,nestin적표체량여병정진전상일치.안압승고시,nestin재Müller세포상유도표체,우기재족판처적강렬표체,가능대RGCs구유보호작용.Nestin유망성위일충신적연구시망막손상적생물표기물.
Background Elevated intraocular pressure leads to the loss of retinal ganglion cells and vigorous reaction of retinal glial cells.The expression of nestin in retinal glial cells secondary to hypertention and its significance are unclear.ObjectiveThis study aim to investigate the expression of nestin in retinal glial cells (RGCs) in ocular hypertention rats.Methods The ocular hypertention models were established by cauterizing the limbus-draining veins in the right eyes of 42 SD rats,and a conjunctival incision in the left eyes of the rats served as the sham group.The intraocular pressure (IOP) was measured with the Tono-Pen XL tonometer.The number of RGCs in the rats with ocular hypertention was counted.The expression of the nestin protein in RGCs was semi-quantitatively analyzed using Western by immunochemistry.Double immunofluorescence was carried out to evaluate the the confocal laser scaning microscope.Results Significant differences were found in the IOP between the model group and the sham group at various time points (P＜0.05).In 1 week to 3 weeks after operation,the number of RGCs significantly declined in the model group compared with the sham group (P＜0.05).Immunochemistry showed that from 2 hours through 1 week after operation,the expression of nestin was gradually enhanced in the model group in comparison with the sham group.Western blot revealed that the expression of the nestin protein reflected a similar tendency to that of immunofluorescence.The increased introcular pressure as manifested by the induced expression of nestin.Immunoelectron microscopy also confirmed the induced expression of nestin especially at their end-feet suggests a potential neuroprotective mechanism in neuronal degeneration.Nestin may be a useful biomarker for retinal injury study.