中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
5期
433-436
,共4页
武劲圆%孙丰源%唐东润%张蕊
武勁圓%孫豐源%唐東潤%張蕊
무경원%손봉원%당동윤%장예
睫状神经营养因子%高眼压%视网膜神经节细胞%谷氨酸
睫狀神經營養因子%高眼壓%視網膜神經節細胞%穀氨痠
첩상신경영양인자%고안압%시망막신경절세포%곡안산
Ciliary neurotrophie factor%Ocular hypertention%Retinal ganglion cell%Glutamic acid
背景 青光眼可以引起视网膜神经节细胞(RGCs)凋亡,据报道睫状神经营养因子(CNTF)对外伤性视神经损伤有修复作用,其是否对青光眼视神经病变有保护作用尚少见报道. 目的 观察CNTF对大鼠急性高眼压眼RGCs的保护作用.方法 24只Wistar大鼠双眼采用眼前房平衡盐液加压灌注法建立大鼠急性高眼压模型,造模前2d左眼玻璃体内注入0.5μg CNTF 5μl,右眼以同样的方法注射磷酸钠溶液5μl,另取3只正常大鼠作为正常对照.造模后1、3、7、14 d过量麻醉法处死动物并摘除眼球,制备视网膜组织学切片后采用苏木精-伊红染色法进行形态学观察,光学显微镜下计数RGCs数目；采用免疫组织化学染色法观察RGCs层谷氨酸的表达情况.结果 正常对照组大鼠视网膜各层排列整齐,细胞边界清晰；模型对照组大鼠RGCs细胞膜、细胞核均发现异常改变,有细胞空泡样变；CNTF治疗组大鼠造模后变性的RGCs数量少.与模型对照组比较,造模后3、7、14 d CNTF治疗组RGCs数目明显增加,差异均有统计学意义(均P=0.000).免疫组织化学染色表明,造模后3～7d,CNTF治疗组RGCs层谷氨酸阳性细胞数分别为(5.50±1.04)个／3个高倍视野和(6.00±1.41) 个/3个高倍视野,明显低于模型对照组的(9.00±2.91)个／3个高倍视野和(10.83±1.94)个／3个高倍视野,差异均有统计学意义(均P=0.000),而造模后1d和14 d两组间谷氨酸阳性细胞数的差异均无统计学意义(P=0.578、0.180).结论 CNTF能够下调急性高眼压眼谷氨酸在RGCs中的表达,从而对RGCs提供保护作用.
揹景 青光眼可以引起視網膜神經節細胞(RGCs)凋亡,據報道睫狀神經營養因子(CNTF)對外傷性視神經損傷有脩複作用,其是否對青光眼視神經病變有保護作用尚少見報道. 目的 觀察CNTF對大鼠急性高眼壓眼RGCs的保護作用.方法 24隻Wistar大鼠雙眼採用眼前房平衡鹽液加壓灌註法建立大鼠急性高眼壓模型,造模前2d左眼玻璃體內註入0.5μg CNTF 5μl,右眼以同樣的方法註射燐痠鈉溶液5μl,另取3隻正常大鼠作為正常對照.造模後1、3、7、14 d過量痳醉法處死動物併摘除眼毬,製備視網膜組織學切片後採用囌木精-伊紅染色法進行形態學觀察,光學顯微鏡下計數RGCs數目；採用免疫組織化學染色法觀察RGCs層穀氨痠的錶達情況.結果 正常對照組大鼠視網膜各層排列整齊,細胞邊界清晰；模型對照組大鼠RGCs細胞膜、細胞覈均髮現異常改變,有細胞空泡樣變；CNTF治療組大鼠造模後變性的RGCs數量少.與模型對照組比較,造模後3、7、14 d CNTF治療組RGCs數目明顯增加,差異均有統計學意義(均P=0.000).免疫組織化學染色錶明,造模後3～7d,CNTF治療組RGCs層穀氨痠暘性細胞數分彆為(5.50±1.04)箇／3箇高倍視野和(6.00±1.41) 箇/3箇高倍視野,明顯低于模型對照組的(9.00±2.91)箇／3箇高倍視野和(10.83±1.94)箇／3箇高倍視野,差異均有統計學意義(均P=0.000),而造模後1d和14 d兩組間穀氨痠暘性細胞數的差異均無統計學意義(P=0.578、0.180).結論 CNTF能夠下調急性高眼壓眼穀氨痠在RGCs中的錶達,從而對RGCs提供保護作用.
배경 청광안가이인기시망막신경절세포(RGCs)조망,거보도첩상신경영양인자(CNTF)대외상성시신경손상유수복작용,기시부대청광안시신경병변유보호작용상소견보도. 목적 관찰CNTF대대서급성고안압안RGCs적보호작용.방법 24지Wistar대서쌍안채용안전방평형염액가압관주법건립대서급성고안압모형,조모전2d좌안파리체내주입0.5μg CNTF 5μl,우안이동양적방법주사린산납용액5μl,령취3지정상대서작위정상대조.조모후1、3、7、14 d과량마취법처사동물병적제안구,제비시망막조직학절편후채용소목정-이홍염색법진행형태학관찰,광학현미경하계수RGCs수목；채용면역조직화학염색법관찰RGCs층곡안산적표체정황.결과 정상대조조대서시망막각층배렬정제,세포변계청석；모형대조조대서RGCs세포막、세포핵균발현이상개변,유세포공포양변；CNTF치료조대서조모후변성적RGCs수량소.여모형대조조비교,조모후3、7、14 d CNTF치료조RGCs수목명현증가,차이균유통계학의의(균P=0.000).면역조직화학염색표명,조모후3～7d,CNTF치료조RGCs층곡안산양성세포수분별위(5.50±1.04)개／3개고배시야화(6.00±1.41) 개/3개고배시야,명현저우모형대조조적(9.00±2.91)개／3개고배시야화(10.83±1.94)개／3개고배시야,차이균유통계학의의(균P=0.000),이조모후1d화14 d량조간곡안산양성세포수적차이균무통계학의의(P=0.578、0.180).결론 CNTF능구하조급성고안압안곡안산재RGCs중적표체,종이대RGCs제공보호작용.
Background Glaucoma associates with apoptosis of retinal ganglion cells ( RGCs).Research showed that ciliary neurotrophic factor (CNTF) can repair optic nerve trauma,but it has not reported whether CNTF has a protective effect on glaucomatous optic neuropathy. Objective This experimental study was to explore the protective effect of CNTF on RGCs in rat eyes with acute ocular hypertension. Methods Ocular acute hypertension models were induced in bilateral eyes of 24 clean Wistar rats by forced perfusion of a balanced salt solution into the anterior chamber.Two days before molding,0.5 μg recombinant human CNTF (5 μl) was injected intravitreously in the left eyes,and 5 mmol/L sodium phosphate solution (5 μl) was injected in the same way as the control group.Three other Wistar rats were used as the normal control group.The animals were sacrificed by excessive anesthesia and the retinal sections were prepared 1,3,7,14 days after molding.The morphology of the retina was examined and RGCs counting was performed by hematoxylin & eosin staining and observed under a light microscope.The expression of glutamic acid in RGCs was assessed by immunochemistry. Results Regular retinal structure with clearly defined cell layers were observed in normal control rats.Changes in vacuoles in RGCs were seen in the model control group.The number of degenerative RGCs decreased in the CNTF group.Compared with the model control group,number of normal RGCs increased from 1 day to 14 days after molding with a significant difference between the two groups (all P=0.000).Immunochemistry assay showed that the numbers of positive cells for glutamic acid were 5.50±1.04 and 6.00±1.41 for the average of 3 fields at 3 days or 7 days in the CNTF group,and those in the model control group were 9.00±2.91 and 10.83 ± 1.94,respectively,showing significant differences between them ( all P =0.000 ).However,no comparable differences were found in the numbers of positive cells for glutamic acid at 1 day and 14 days between the two groups( P=0.578,0.180). Conclusions CNTF down-regulates the expression of glutamic acid in RGCs and offers neuroprotection for RGCs in an acute glaucoma rat model.
