中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2008年
5期
391-397
,共7页
高眼压%疾病模型%动物%视网膜%神经胶质%视网膜神经节细胞
高眼壓%疾病模型%動物%視網膜%神經膠質%視網膜神經節細胞
고안압%질병모형%동물%시망막%신경효질%시망막신경절세포
Ocular hypertension%Disease models,animal%Retinal%Neuroglia%Retinal ganglion cells
目的 研究青光眼视网膜神经胶质细胞组织病理学改变及其在青光眼视网膜神经节细胞损伤中的作用机制.方法 对照实验研究.选用造模成功的慢性高眼压雄性SD大鼠72只,眼压＞22 mm Hg(1 mm Hg=0.133 kPa).右眼为慢性高眼压眼,左眼为假手术对照眼.根据慢性高眼压模型建立的时间(手术结束时开始计算),将实验鼠分为6组(2、12 h,1 d,1、4、8周),每组12只鼠.正常组SD大鼠12只,平均眼压12.56 mm Hg.分别取实验各组(慢性高眼压)、假手术组及正常组大鼠4只眼球,冰冻切片行胶质纤维酸性蛋白(GFAP)免疫组织化学染色,在激光共焦显微镜下观察视网膜星形胶质细胞及Müller细胞的GFAP表达情况;分别取实验各组(慢性高眼压)和正常组大鼠4只眼球,在视网膜铺片上进行GFAP免疫组织化学染色,进行星形胶质细胞计数并观察其形态;分别取实验各组(慢性高眼压)、假手术组及正常大鼠4只眼球的鼻侧半视网膜,在视网膜铺片上行小胶质细胞标记物OX42免疫组织化学染色,进行小胶质细胞计数并观察其形态;取剩余的颢侧中周部视网膜,半薄切片行甲苯胺蓝染色并进行Müller细胞计数.对不同时间点慢性高眼压组与正常组大鼠细胞表达数最进行比较,采用单因素多水平设计定量资料的方差分析.结果 慢性高眼压模型建立后2 h,即有活化的小胶质细胞出现;1 d后小胶质细胞的数量开始增加,为(327.40±68.32)个/mm2;1周后小胶质细胞的数量达到高峰,为(965.06±86.63)个/mmw,与正常组小胶质细胞数最比较,差异有统计学意义(F=196.56,P＜0.01);其后小胶质细胞数量逐渐减少.慢性高眼压模型建立后12 h,星形胶质细胞及Müller细胞开始呈现活化状态;4周时两种细胞的活化程度达到高峰,以后活化程度逐渐下降,且活化的星形胶质细胞在结构上出现明显异常,表现为星形胶质细胞突起变得粗短、僵硬,胞体的星型结构破坏;但慢性高眼压组视网膜星形胶质细胞及Müller细胞数量与正常组相比,差异均无统计学意义(F=1.36,1.89;均P＞0.05).结论 在慢性高眼压条件下,小胶质细胞可能是视网膜最早发生病理学改变的组织;活化的星形胶质细胞可出现明显的形态和结构变化,其结果不仅将加速神经节细胞的损伤,同时也会形成不利于神经节细胞轴突再生的视网膜微环境.
目的 研究青光眼視網膜神經膠質細胞組織病理學改變及其在青光眼視網膜神經節細胞損傷中的作用機製.方法 對照實驗研究.選用造模成功的慢性高眼壓雄性SD大鼠72隻,眼壓＞22 mm Hg(1 mm Hg=0.133 kPa).右眼為慢性高眼壓眼,左眼為假手術對照眼.根據慢性高眼壓模型建立的時間(手術結束時開始計算),將實驗鼠分為6組(2、12 h,1 d,1、4、8週),每組12隻鼠.正常組SD大鼠12隻,平均眼壓12.56 mm Hg.分彆取實驗各組(慢性高眼壓)、假手術組及正常組大鼠4隻眼毬,冰凍切片行膠質纖維痠性蛋白(GFAP)免疫組織化學染色,在激光共焦顯微鏡下觀察視網膜星形膠質細胞及Müller細胞的GFAP錶達情況;分彆取實驗各組(慢性高眼壓)和正常組大鼠4隻眼毬,在視網膜鋪片上進行GFAP免疫組織化學染色,進行星形膠質細胞計數併觀察其形態;分彆取實驗各組(慢性高眼壓)、假手術組及正常大鼠4隻眼毬的鼻側半視網膜,在視網膜鋪片上行小膠質細胞標記物OX42免疫組織化學染色,進行小膠質細胞計數併觀察其形態;取剩餘的顥側中週部視網膜,半薄切片行甲苯胺藍染色併進行Müller細胞計數.對不同時間點慢性高眼壓組與正常組大鼠細胞錶達數最進行比較,採用單因素多水平設計定量資料的方差分析.結果 慢性高眼壓模型建立後2 h,即有活化的小膠質細胞齣現;1 d後小膠質細胞的數量開始增加,為(327.40±68.32)箇/mm2;1週後小膠質細胞的數量達到高峰,為(965.06±86.63)箇/mmw,與正常組小膠質細胞數最比較,差異有統計學意義(F=196.56,P＜0.01);其後小膠質細胞數量逐漸減少.慢性高眼壓模型建立後12 h,星形膠質細胞及Müller細胞開始呈現活化狀態;4週時兩種細胞的活化程度達到高峰,以後活化程度逐漸下降,且活化的星形膠質細胞在結構上齣現明顯異常,錶現為星形膠質細胞突起變得粗短、僵硬,胞體的星型結構破壞;但慢性高眼壓組視網膜星形膠質細胞及Müller細胞數量與正常組相比,差異均無統計學意義(F=1.36,1.89;均P＞0.05).結論 在慢性高眼壓條件下,小膠質細胞可能是視網膜最早髮生病理學改變的組織;活化的星形膠質細胞可齣現明顯的形態和結構變化,其結果不僅將加速神經節細胞的損傷,同時也會形成不利于神經節細胞軸突再生的視網膜微環境.
목적 연구청광안시망막신경효질세포조직병이학개변급기재청광안시망막신경절세포손상중적작용궤제.방법 대조실험연구.선용조모성공적만성고안압웅성SD대서72지,안압＞22 mm Hg(1 mm Hg=0.133 kPa).우안위만성고안압안,좌안위가수술대조안.근거만성고안압모형건립적시간(수술결속시개시계산),장실험서분위6조(2、12 h,1 d,1、4、8주),매조12지서.정상조SD대서12지,평균안압12.56 mm Hg.분별취실험각조(만성고안압)、가수술조급정상조대서4지안구,빙동절편행효질섬유산성단백(GFAP)면역조직화학염색,재격광공초현미경하관찰시망막성형효질세포급Müller세포적GFAP표체정황;분별취실험각조(만성고안압)화정상조대서4지안구,재시망막포편상진행GFAP면역조직화학염색,진행성형효질세포계수병관찰기형태;분별취실험각조(만성고안압)、가수술조급정상대서4지안구적비측반시망막,재시망막포편상행소효질세포표기물OX42면역조직화학염색,진행소효질세포계수병관찰기형태;취잉여적호측중주부시망막,반박절편행갑분알람염색병진행Müller세포계수.대불동시간점만성고안압조여정상조대서세포표체수최진행비교,채용단인소다수평설계정량자료적방차분석.결과 만성고안압모형건립후2 h,즉유활화적소효질세포출현;1 d후소효질세포적수량개시증가,위(327.40±68.32)개/mm2;1주후소효질세포적수량체도고봉,위(965.06±86.63)개/mmw,여정상조소효질세포수최비교,차이유통계학의의(F=196.56,P＜0.01);기후소효질세포수량축점감소.만성고안압모형건립후12 h,성형효질세포급Müller세포개시정현활화상태;4주시량충세포적활화정도체도고봉,이후활화정도축점하강,차활화적성형효질세포재결구상출현명현이상,표현위성형효질세포돌기변득조단、강경,포체적성형결구파배;단만성고안압조시망막성형효질세포급Müller세포수량여정상조상비,차이균무통계학의의(F=1.36,1.89;균P＞0.05).결론 재만성고안압조건하,소효질세포가능시시망막최조발생병이학개변적조직;활화적성형효질세포가출현명현적형태화결구변화,기결과불부장가속신경절세포적손상,동시야회형성불리우신경절세포축돌재생적시망막미배경.
Objective To study the pathological changes of retinal glial cells and its effect on retinal ganglion cells (RGC) damage in a rat chronic ocular hypertension model. Methods Seventy-two of Sprague-Dawley (SD) rats with chronic elevated intraocular pressure (IOP) by ligating two superior or inferior episcleral veins were used in this study. Twelve normal rats served as control. The densities of glial cells were determined in flat mounted or transverse semithin sections of retinas, microglial cells were visualized by OX42 staining on whole-mounted retinas, and Muller cells were detected by expressing glial fibrillary acidic protein (GFAP) on frozen section or flat mounted retinas with confocal microscopy at 2 hours, 12 hours, 1 day, 1 week, 4 weeks, and 8 weeks after operation, respectively. Results Compared with control group, the densities of activated microglial cells were significantly ( F = 196. 56, P < 0. 01 )increased at 1 day (215.00±18.60 vs 327. 40±68. 32/mm2) and reached a peak value at 1 week ( 965.06±86. 63/mm2), then decreased gradually. But the densities of astrocytes and Muller cells were notsignificantly changed (F = 1.36, 1.89 ; P>0. 05 ) at all time points. The activated microglial cells were appeared at 2 hour while the activated Muller cells and astrocytes presented at 12 hours after operation. The activated Muller cells and astrocytes reached a peak value at 4 weeks, and then decreased gradually. The activated astrocyte had morphological changes including disappeared star-structure of cell body, stiffness,and shortness of cell processes. Conclusions The alteration of microglial cell densities appears to be the earliest pathological changes of retina in rat with chronic ocular hypertension. The activated astrocytes with morphological disorder may deteriorate the damage of RGC and result in a harmful microenvironment for axonal regeneration of RGC in glaucoma.
