中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2012年
3期
272-275
,共4页
闫明%宋艳萍%李付亮%陈中山
閆明%宋豔萍%李付亮%陳中山
염명%송염평%리부량%진중산
视网膜变性/病理生理学%视网膜神经节细胞/生理学%钙/副作用%动物实验
視網膜變性/病理生理學%視網膜神經節細胞/生理學%鈣/副作用%動物實驗
시망막변성/병리생이학%시망막신경절세포/생이학%개/부작용%동물실험
Retinal degeneration/pathophysiology%Retinal ganglion cells/physiology%Calcium/adverse effects%Animal experimentation
目的 观察探讨视网膜变性大鼠视网膜神经节细胞(RGCs)的变化及其触发因素.方法 出生后21、60、90 d含色素的Royal College of Surgeons(RCS)-p+视网膜变性大鼠各6只,RCS-rdy+ -p+无视网膜变性大鼠各6只.立体定位下行大鼠上丘和外膝体荧光金逆行标记RGCs,荧光显微镜下观察RGCs细胞形态并计数.作视网膜铺片和切片,采用激光共聚焦显微镜观察RGCs细胞内钙成像情况,分析RGCs细胞内钙浓度的变化.结果 荧光显微镜观察发现,出生后90 d,无视网膜变性大鼠RGCs分布于整个视网膜,细胞大小均一,细胞胞体和突起清晰可见；视网膜变性大鼠RGCs分布稀疏,细胞大小不一,细胞突起分枝数和范围减小,出现杆状或碎屑细胞.出生后21、60、90 d,视网膜变性大鼠RGCs数量分别为(5421.0±72.1)、(4195.0±136.4)、(2906.0±133.2)个/mm2.与无视网膜变性大鼠比较,出生后21d时其RGCs数量间差异无统计学意义(t=-1.301,P＞0.05)；出生后60、90 d时其RGCs数量显著减少,差异有统计学意义(t=16.172,30.131；P＜0.05).出生后21d有无视网膜变性大鼠RGCs细胞内钙荧光强度比较,差异无统计学意义(t=-1.545,P＞0.05)；出生后60、90 d视网膜变性大鼠RGCs细胞内钙荧光强度较无视网膜变性大鼠明显增强,差异有统计学意义(t=-18.058,-15.015；P＜0.0l).结论 视网膜变性大鼠RGCs发生变性死亡,细胞形态和数量受到显著影响.RGCs细胞内钙浓度的变化可能是视网膜变性中RGCs继发变性、死亡的触发因素.
目的 觀察探討視網膜變性大鼠視網膜神經節細胞(RGCs)的變化及其觸髮因素.方法 齣生後21、60、90 d含色素的Royal College of Surgeons(RCS)-p+視網膜變性大鼠各6隻,RCS-rdy+ -p+無視網膜變性大鼠各6隻.立體定位下行大鼠上丘和外膝體熒光金逆行標記RGCs,熒光顯微鏡下觀察RGCs細胞形態併計數.作視網膜鋪片和切片,採用激光共聚焦顯微鏡觀察RGCs細胞內鈣成像情況,分析RGCs細胞內鈣濃度的變化.結果 熒光顯微鏡觀察髮現,齣生後90 d,無視網膜變性大鼠RGCs分佈于整箇視網膜,細胞大小均一,細胞胞體和突起清晰可見；視網膜變性大鼠RGCs分佈稀疏,細胞大小不一,細胞突起分枝數和範圍減小,齣現桿狀或碎屑細胞.齣生後21、60、90 d,視網膜變性大鼠RGCs數量分彆為(5421.0±72.1)、(4195.0±136.4)、(2906.0±133.2)箇/mm2.與無視網膜變性大鼠比較,齣生後21d時其RGCs數量間差異無統計學意義(t=-1.301,P＞0.05)；齣生後60、90 d時其RGCs數量顯著減少,差異有統計學意義(t=16.172,30.131；P＜0.05).齣生後21d有無視網膜變性大鼠RGCs細胞內鈣熒光彊度比較,差異無統計學意義(t=-1.545,P＞0.05)；齣生後60、90 d視網膜變性大鼠RGCs細胞內鈣熒光彊度較無視網膜變性大鼠明顯增彊,差異有統計學意義(t=-18.058,-15.015；P＜0.0l).結論 視網膜變性大鼠RGCs髮生變性死亡,細胞形態和數量受到顯著影響.RGCs細胞內鈣濃度的變化可能是視網膜變性中RGCs繼髮變性、死亡的觸髮因素.
목적 관찰탐토시망막변성대서시망막신경절세포(RGCs)적변화급기촉발인소.방법 출생후21、60、90 d함색소적Royal College of Surgeons(RCS)-p+시망막변성대서각6지,RCS-rdy+ -p+무시망막변성대서각6지.입체정위하행대서상구화외슬체형광금역행표기RGCs,형광현미경하관찰RGCs세포형태병계수.작시망막포편화절편,채용격광공취초현미경관찰RGCs세포내개성상정황,분석RGCs세포내개농도적변화.결과 형광현미경관찰발현,출생후90 d,무시망막변성대서RGCs분포우정개시망막,세포대소균일,세포포체화돌기청석가견；시망막변성대서RGCs분포희소,세포대소불일,세포돌기분지수화범위감소,출현간상혹쇄설세포.출생후21、60、90 d,시망막변성대서RGCs수량분별위(5421.0±72.1)、(4195.0±136.4)、(2906.0±133.2)개/mm2.여무시망막변성대서비교,출생후21d시기RGCs수량간차이무통계학의의(t=-1.301,P＞0.05)；출생후60、90 d시기RGCs수량현저감소,차이유통계학의의(t=16.172,30.131；P＜0.05).출생후21d유무시망막변성대서RGCs세포내개형광강도비교,차이무통계학의의(t=-1.545,P＞0.05)；출생후60、90 d시망막변성대서RGCs세포내개형광강도교무시망막변성대서명현증강,차이유통계학의의(t=-18.058,-15.015；P＜0.0l).결론 시망막변성대서RGCs발생변성사망,세포형태화수량수도현저영향.RGCs세포내개농도적변화가능시시망막변성중RGCs계발변성、사망적촉발인소.
Objective To investigate the triggering mechanisms of retinal ganglion cells (RGCs)alterations in retinally degenerative rats.Methods Retinal dystrophic Royal College of Surgeons (RCS-p+)and control RCS rats (RCS-rdy+-p+) were divided into three groups according to postnatal days,including postnatal 21 days (P21d),P60d and P90d,with six rats in each group. Fluorogold was injected into superior colliculus and lateral geniculate body for retrograde labeling RGCs.Then retinal flat mounts were observed under fluorescence microscope to investigate the morphological and RGC changes in density during retinal degeneration.Living retinal mounts or sections were prepared to investigate the calcium concentration of RGCs using a laser confocal microscope. Results At P90d RGCs of control rats were distributed throughout entire retina and appeared uniform.The soma and processes were apparent and clear.But at P90d RGCs of RCS-p+ rats were distributed sparsely in retina.The soma seemed messy and the number and dendritic fields decreased greatly,and some baculiform or broken cells appeared. RGC density was5421.0±72.1/mm2 at P21d,4195.0±136.4/mm2 at P60dand2906.0±133.2/mm2 at P90d.At P21d there were no obvious differences in RGCs density between RCS-p+ and control rats (t=-1.301,P＞0.05).At P60d and P90d there were significant differences in RGC density (t=16.172,30.131; P＜0.01).At P2ld there was no significant differences in RGC intracellular calcium fluorescence intensity between RCS-p+ and control rats (t=-1.545,P＞0.05).At P60d and P90d the intracellular calcium fluorescence intensity decreased obviously and the differences were significant (t=-18.058,-15.015; P＜0.01).Conclusions RGCs were secondarily impaired following the loss of photoreceptors at late stages of retinal degeneration,and the morphological and cell density of RGCs were affected.The overloading of intracellular calcium concentration may be the triggering factor of degeneration and death of RGCs in retinally dystrophic rats.
