中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2014年
2期
143-148
,共6页
晶状体/损伤%外伤性白内障%视神经/横断伤%Müller细胞%视网膜神经节细胞%细胞存活/生理功能
晶狀體/損傷%外傷性白內障%視神經/橫斷傷%Müller細胞%視網膜神經節細胞%細胞存活/生理功能
정상체/손상%외상성백내장%시신경/횡단상%Müller세포%시망막신경절세포%세포존활/생리공능
Lens/injury%Cataractogenic lens injury%Optic nerve/axotomy%M(u)ller cell%Retinal ganglion cell%Cell survival/physiology
背景 大鼠Müller细胞提取液能够促进离体培养的视网膜神经节细胞(RGCs)存活及轴突的再生,伴晶状体损伤的视神经外伤眼RGCs存活率提高,但Müller细胞和晶状体损伤在促进RGCs存活方面的关系鲜见报道.目的 探讨伴晶状体损伤的视神经外伤眼Müller细胞对RGCs存活的促进作用及其机制.方法清洁级成年Wistar大鼠48只按随机数字表法随机分为伪手术组、视神经损伤组、晶状体联合视神经损伤组.伪手术组大鼠手术中暴露但不损伤视神经,视神经损伤组大鼠行视神经横断伤,晶状体联合视神经损伤组行视神经横断伤联合晶状体针刺伤,并导致晶状体混浊.术后7d及14 d各组分别取8只大鼠处死后制备视网膜标本.采用苏木精-伊红染色观察各组大鼠视网膜和RGCs的形态学改变,采用免疫组织化学法检测各组大鼠视网膜内核层胶质纤维酸性蛋白(GFAP)标记的Müller细胞,光学显微镜下计数各组大鼠RGCs数量及GFAP阳性标记的Müller细胞数量. 结果 术后7d及14 d,伪手术组大鼠RGCs的数量分别为(52.98±1.90)个/高倍视野和(51.81±3.09)个/高倍视野,差异无统计学意义(t=0.910,P=0.378)；术后14d视神经损伤组大鼠RGCs数量为(22.67±1.94)个/高倍视野,明显少于术后7d的(36.61±1.69)个/高倍视野,差异有统计学意义(t=15.312,P=0.000)；术后14 d晶状体联合视神经损伤组RGCs数量为(35.69±1.80)个/高倍视野,明显少于术后7d的(50.76±2.77)个/高倍视野,差异有统计学意义(t=12.920,P=0.000).术后7d及14d,晶状体联合视神经损伤组存活的RGCs数量均多于视神经损伤组,差异均有统计学意义(7 d:扛102.840,P=0.000；14 d:t=164.020,P=0.000)；术后14d晶状体联合视神经损伤组存活的RGCs数量少于伪手术组,差异有统计学意义(t=187.04,P=0.034).术后7d及14d,伪手术组大鼠视网膜内核层均未见GFAP阳性标记的Müller细胞；视神经损伤组大鼠内核层GFAP阳性标记Müller细胞数量分别为(29.38±2.04)个/高倍视野和(19.07±2.14)个/高倍视野,差异有统计学意义(t=-9.868,P=0.000).晶状体联合视神经损伤组大鼠内核层GFAP阳性标记的Müller细胞数量分别为(48.96±2.80)个/高倍视野和(46.73±1.50)个/高倍视野,差异无统计学意义(t=1.987,P=0.067).术后7d及14d,晶状体联合视神经损伤组大鼠内核层GFAP阳性Müller细胞数量均较视神经损伤组增多,差异均有统计学意义(7d:t=-15.997,P=0.000；14 d:t=-29.938,P=0.000). 结论 在视神经损伤合并晶状体刺伤时,晶状体损伤可诱导Müller细胞活化,进而促进视神经损伤后RGCs的存活.
揹景 大鼠Müller細胞提取液能夠促進離體培養的視網膜神經節細胞(RGCs)存活及軸突的再生,伴晶狀體損傷的視神經外傷眼RGCs存活率提高,但Müller細胞和晶狀體損傷在促進RGCs存活方麵的關繫鮮見報道.目的 探討伴晶狀體損傷的視神經外傷眼Müller細胞對RGCs存活的促進作用及其機製.方法清潔級成年Wistar大鼠48隻按隨機數字錶法隨機分為偽手術組、視神經損傷組、晶狀體聯閤視神經損傷組.偽手術組大鼠手術中暴露但不損傷視神經,視神經損傷組大鼠行視神經橫斷傷,晶狀體聯閤視神經損傷組行視神經橫斷傷聯閤晶狀體針刺傷,併導緻晶狀體混濁.術後7d及14 d各組分彆取8隻大鼠處死後製備視網膜標本.採用囌木精-伊紅染色觀察各組大鼠視網膜和RGCs的形態學改變,採用免疫組織化學法檢測各組大鼠視網膜內覈層膠質纖維痠性蛋白(GFAP)標記的Müller細胞,光學顯微鏡下計數各組大鼠RGCs數量及GFAP暘性標記的Müller細胞數量. 結果 術後7d及14 d,偽手術組大鼠RGCs的數量分彆為(52.98±1.90)箇/高倍視野和(51.81±3.09)箇/高倍視野,差異無統計學意義(t=0.910,P=0.378)；術後14d視神經損傷組大鼠RGCs數量為(22.67±1.94)箇/高倍視野,明顯少于術後7d的(36.61±1.69)箇/高倍視野,差異有統計學意義(t=15.312,P=0.000)；術後14 d晶狀體聯閤視神經損傷組RGCs數量為(35.69±1.80)箇/高倍視野,明顯少于術後7d的(50.76±2.77)箇/高倍視野,差異有統計學意義(t=12.920,P=0.000).術後7d及14d,晶狀體聯閤視神經損傷組存活的RGCs數量均多于視神經損傷組,差異均有統計學意義(7 d:扛102.840,P=0.000；14 d:t=164.020,P=0.000)；術後14d晶狀體聯閤視神經損傷組存活的RGCs數量少于偽手術組,差異有統計學意義(t=187.04,P=0.034).術後7d及14d,偽手術組大鼠視網膜內覈層均未見GFAP暘性標記的Müller細胞；視神經損傷組大鼠內覈層GFAP暘性標記Müller細胞數量分彆為(29.38±2.04)箇/高倍視野和(19.07±2.14)箇/高倍視野,差異有統計學意義(t=-9.868,P=0.000).晶狀體聯閤視神經損傷組大鼠內覈層GFAP暘性標記的Müller細胞數量分彆為(48.96±2.80)箇/高倍視野和(46.73±1.50)箇/高倍視野,差異無統計學意義(t=1.987,P=0.067).術後7d及14d,晶狀體聯閤視神經損傷組大鼠內覈層GFAP暘性Müller細胞數量均較視神經損傷組增多,差異均有統計學意義(7d:t=-15.997,P=0.000；14 d:t=-29.938,P=0.000). 結論 在視神經損傷閤併晶狀體刺傷時,晶狀體損傷可誘導Müller細胞活化,進而促進視神經損傷後RGCs的存活.
배경 대서Müller세포제취액능구촉진리체배양적시망막신경절세포(RGCs)존활급축돌적재생,반정상체손상적시신경외상안RGCs존활솔제고,단Müller세포화정상체손상재촉진RGCs존활방면적관계선견보도.목적 탐토반정상체손상적시신경외상안Müller세포대RGCs존활적촉진작용급기궤제.방법청길급성년Wistar대서48지안수궤수자표법수궤분위위수술조、시신경손상조、정상체연합시신경손상조.위수술조대서수술중폭로단불손상시신경,시신경손상조대서행시신경횡단상,정상체연합시신경손상조행시신경횡단상연합정상체침자상,병도치정상체혼탁.술후7d급14 d각조분별취8지대서처사후제비시망막표본.채용소목정-이홍염색관찰각조대서시망막화RGCs적형태학개변,채용면역조직화학법검측각조대서시망막내핵층효질섬유산성단백(GFAP)표기적Müller세포,광학현미경하계수각조대서RGCs수량급GFAP양성표기적Müller세포수량. 결과 술후7d급14 d,위수술조대서RGCs적수량분별위(52.98±1.90)개/고배시야화(51.81±3.09)개/고배시야,차이무통계학의의(t=0.910,P=0.378)；술후14d시신경손상조대서RGCs수량위(22.67±1.94)개/고배시야,명현소우술후7d적(36.61±1.69)개/고배시야,차이유통계학의의(t=15.312,P=0.000)；술후14 d정상체연합시신경손상조RGCs수량위(35.69±1.80)개/고배시야,명현소우술후7d적(50.76±2.77)개/고배시야,차이유통계학의의(t=12.920,P=0.000).술후7d급14d,정상체연합시신경손상조존활적RGCs수량균다우시신경손상조,차이균유통계학의의(7 d:강102.840,P=0.000；14 d:t=164.020,P=0.000)；술후14d정상체연합시신경손상조존활적RGCs수량소우위수술조,차이유통계학의의(t=187.04,P=0.034).술후7d급14d,위수술조대서시망막내핵층균미견GFAP양성표기적Müller세포；시신경손상조대서내핵층GFAP양성표기Müller세포수량분별위(29.38±2.04)개/고배시야화(19.07±2.14)개/고배시야,차이유통계학의의(t=-9.868,P=0.000).정상체연합시신경손상조대서내핵층GFAP양성표기적Müller세포수량분별위(48.96±2.80)개/고배시야화(46.73±1.50)개/고배시야,차이무통계학의의(t=1.987,P=0.067).술후7d급14d,정상체연합시신경손상조대서내핵층GFAP양성Müller세포수량균교시신경손상조증다,차이균유통계학의의(7d:t=-15.997,P=0.000；14 d:t=-29.938,P=0.000). 결론 재시신경손상합병정상체자상시,정상체손상가유도Müller세포활화,진이촉진시신경손상후RGCs적존활.
Background It has been reported that murine Müller cells conditional medium can promote the survival of retinal ganglion cells (RGCs) and the regeneration of axons,and the survival rate of RGCs improve in the optic nerve axotomy eyes with cataractogenic lens injury in vitro.However,the interaction of Müller cells with pricking of lens in protecting RGCs is unclear.Objective The aim of this study was to investigate the role of Müller cells on survival of RGCs in the optic nerve axotomy with cataractogenic lens injury.Methods Forty-eight clean adult Wistar rats were randomized into sham operation group,optic nerve axotomy group and lens injury combined with optic nerve axotomy group.The optic nerve was exposed only in the rats of the sham operation group,optic nerve was completely transected at 3 mm behind the eyeball in the rats of the optic nerve axotomy group,and lens puncture and optic nerve axotomy were performed in the eyes of lens injury combined with optic nerve axotomy group.The rats were sacrificed at day 7 and day 14 after operation to prepare the retinal specimens.The RGCs were examined and counted by hematoxylin-eosin staining.Müller cells labeled by glial fibrillary acidic protein (GFAP) were counted using immunohistochemisty.Results The number of RGCs was (52.98 ± 1.90) /field and (51.81 ±3.09) /field on the 7th and 14th day in the sham operation group,without significant difference between them (t =0.910,P =0.378).The number of RGCs was significantly lower on the 14th day ([22.67±1.94] /field) than that of the 7th day ([36.61±1.69] /field) in the optic nerve axotomy group (t=15.312,P=0.000).Also,the number of RGCs was (50.76±2.77) /field and (35.69±1.80) /field on the 7th and 14th day in the lens injury combined with optic nerve axotomy group,showing a significant difference between the two timepoints (t =12.920,P =0.000).In addition,the RGCs number in the lens injury combined with optic nerve axotomy group was significantly higher than that in the optic nerve axotomy group both on 7 days and 14 days after operation (7 days:t =102.840,P =0.000; 14 days:t =164.020,P =0.000),and the number of RGCs was lower in the lens injury combined with optic nerve axotomy group than that of the sham operation group on day 14 (t =187.040,P =0.034).None of GFAP-labeled Müller cell was seen in sham operation group at both on 7 days and 14 days after operation,but a significant difference was found in the optic nerve axotomy group between the two timepoints ([29.38 ± 2.04]/field vs.[19.07 ± 2.14]/field ; t =-9.868,P=0.000).No significant difference in the number of the GFAP-labeled Müller cells was found in the lens injury combined with optic nerve axotomy group between 7 days and 14 days after operation([48.96±2.80] /field vs.[46.73±1.50]/field,t=1.987,P=0.067).In postoperative 7 days and 14 days after operation,the number of GFAP-labeled Müller cells increased in the lens injury combined with optic nerve axotomy group compared with the optic nerve axotomy group (7 days:t =-15.997,P=0.000; 14 days:t=-29.938,P=0.000).Conclusions In optic nerve axotomy with cataractogenic lens injury eye,the punctural injury of lens induce the activity of Müller cells and further promote the survival of RGCs in the cataratogenic lens injury combined with optic nerve axotomy rat eyes.
