中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
6期
505-510
,共6页
张婷%姜利斌%谢君%陈菲%杨冬梅%金玉兰
張婷%薑利斌%謝君%陳菲%楊鼕梅%金玉蘭
장정%강리빈%사군%진비%양동매%금옥란
表没食子儿茶素没食子酸酯%外侧膝状体%神经保护%视神经钳夹伤%神经型一氧化氮合成酶
錶沒食子兒茶素沒食子痠酯%外側膝狀體%神經保護%視神經鉗夾傷%神經型一氧化氮閤成酶
표몰식자인다소몰식자산지%외측슬상체%신경보호%시신경겸협상%신경형일양화담합성매
Epigallocatechin-3-gallate%Lateral geniculate nucleus%Neuroprotection%Optic nerve crush%Neuronal nitric oxide synthase
背景 研究证实绿茶提取物表没食子儿茶素没食子酸酯(EGCG)全身应用可到达视神经及视网膜组织,对视网膜缺血-再灌注和视神经钳夹伤后视网膜神经节细胞(RGCs)具有保护作用,但EGCG对视神经损伤后RGCs上位神经元的影响目前尚未见报道.目的 探讨EGCG对大鼠视神经钳夹伤后外侧膝状体(LGN)神经元的保护作用.方法 48只Wistar大鼠按随机数字表法分为正常对照组、假手术+EGCG组、视神经钳夹+生理盐水组、视神经钳夹+EGCG组,每组12只.用40g微型视神经夹于大鼠右眼球后约2mm处夹持视神经60s建立视神经钳夹伤模型,假手术+EGCG组、视神经钳夹+EGCG组大鼠于造模前2d始每日腹腔内注射EGCG(25mg/kg)共5d,后改为口服(2mg/kg),视神经钳夹+生理盐水组大鼠以同样的方法注射生理盐水.于造模4周后处死大鼠并取脑组织,用Nissl染色法计数外侧膝状体背侧核(dLGN)神经元数目,用免疫组织化学染色法和Western blot法观察神经丝蛋白(NF-L)在LGN的表达,比较各组大鼠dLGN中神经型一氧化氮合酶(nNOS)免疫组织化学染色阳性细胞数量.结果 视神经钳夹伤后4周,假手术+EGCG组左侧、右侧dLGN神经元数量与正常对照组相比差异无统计学意义(P=0.906、P=0.561);视神经钳夹+生理盐水组、视神经钳夹+EGCG组钳夹同侧dLGN神经元数量与正常对照组相比,差异无统计学意义(P=0.794、P=0.646),对侧dLGN神经元数量均低于正常对照组(P=0.000、P=0.015),而视神经钳夹+EGCG组钳夹对侧dLGN神经元数量高于视神经钳夹+生理盐水组(P=0.007);NF-L检测可见视神经钳夹+EGCG组钳夹对侧LGN的NF-L表达量高于视神经钳夹+生理盐水组(P=0.002);dLGN的nNOS阳性细胞计数在正常对照组、假手术+EGCG组、视神经钳夹+EGCG组之间差异无统计学意义(P>0.05),而视神经钳夹+生理盐水组钳夹对侧dLGN的nNOS阳性细胞高于视神经钳夹+EGCG组(P=0.000).结论 EGCG对大鼠视神经钳夹伤后LGN的神经元可能具有一定的保护作用,这种保护作用可能与EGCG抑制了nNOS的表达有关.
揹景 研究證實綠茶提取物錶沒食子兒茶素沒食子痠酯(EGCG)全身應用可到達視神經及視網膜組織,對視網膜缺血-再灌註和視神經鉗夾傷後視網膜神經節細胞(RGCs)具有保護作用,但EGCG對視神經損傷後RGCs上位神經元的影響目前尚未見報道.目的 探討EGCG對大鼠視神經鉗夾傷後外側膝狀體(LGN)神經元的保護作用.方法 48隻Wistar大鼠按隨機數字錶法分為正常對照組、假手術+EGCG組、視神經鉗夾+生理鹽水組、視神經鉗夾+EGCG組,每組12隻.用40g微型視神經夾于大鼠右眼毬後約2mm處夾持視神經60s建立視神經鉗夾傷模型,假手術+EGCG組、視神經鉗夾+EGCG組大鼠于造模前2d始每日腹腔內註射EGCG(25mg/kg)共5d,後改為口服(2mg/kg),視神經鉗夾+生理鹽水組大鼠以同樣的方法註射生理鹽水.于造模4週後處死大鼠併取腦組織,用Nissl染色法計數外側膝狀體揹側覈(dLGN)神經元數目,用免疫組織化學染色法和Western blot法觀察神經絲蛋白(NF-L)在LGN的錶達,比較各組大鼠dLGN中神經型一氧化氮閤酶(nNOS)免疫組織化學染色暘性細胞數量.結果 視神經鉗夾傷後4週,假手術+EGCG組左側、右側dLGN神經元數量與正常對照組相比差異無統計學意義(P=0.906、P=0.561);視神經鉗夾+生理鹽水組、視神經鉗夾+EGCG組鉗夾同側dLGN神經元數量與正常對照組相比,差異無統計學意義(P=0.794、P=0.646),對側dLGN神經元數量均低于正常對照組(P=0.000、P=0.015),而視神經鉗夾+EGCG組鉗夾對側dLGN神經元數量高于視神經鉗夾+生理鹽水組(P=0.007);NF-L檢測可見視神經鉗夾+EGCG組鉗夾對側LGN的NF-L錶達量高于視神經鉗夾+生理鹽水組(P=0.002);dLGN的nNOS暘性細胞計數在正常對照組、假手術+EGCG組、視神經鉗夾+EGCG組之間差異無統計學意義(P>0.05),而視神經鉗夾+生理鹽水組鉗夾對側dLGN的nNOS暘性細胞高于視神經鉗夾+EGCG組(P=0.000).結論 EGCG對大鼠視神經鉗夾傷後LGN的神經元可能具有一定的保護作用,這種保護作用可能與EGCG抑製瞭nNOS的錶達有關.
배경 연구증실록다제취물표몰식자인다소몰식자산지(EGCG)전신응용가도체시신경급시망막조직,대시망막결혈-재관주화시신경겸협상후시망막신경절세포(RGCs)구유보호작용,단EGCG대시신경손상후RGCs상위신경원적영향목전상미견보도.목적 탐토EGCG대대서시신경겸협상후외측슬상체(LGN)신경원적보호작용.방법 48지Wistar대서안수궤수자표법분위정상대조조、가수술+EGCG조、시신경겸협+생리염수조、시신경겸협+EGCG조,매조12지.용40g미형시신경협우대서우안구후약2mm처협지시신경60s건립시신경겸협상모형,가수술+EGCG조、시신경겸협+EGCG조대서우조모전2d시매일복강내주사EGCG(25mg/kg)공5d,후개위구복(2mg/kg),시신경겸협+생리염수조대서이동양적방법주사생리염수.우조모4주후처사대서병취뇌조직,용Nissl염색법계수외측슬상체배측핵(dLGN)신경원수목,용면역조직화학염색법화Western blot법관찰신경사단백(NF-L)재LGN적표체,비교각조대서dLGN중신경형일양화담합매(nNOS)면역조직화학염색양성세포수량.결과 시신경겸협상후4주,가수술+EGCG조좌측、우측dLGN신경원수량여정상대조조상비차이무통계학의의(P=0.906、P=0.561);시신경겸협+생리염수조、시신경겸협+EGCG조겸협동측dLGN신경원수량여정상대조조상비,차이무통계학의의(P=0.794、P=0.646),대측dLGN신경원수량균저우정상대조조(P=0.000、P=0.015),이시신경겸협+EGCG조겸협대측dLGN신경원수량고우시신경겸협+생리염수조(P=0.007);NF-L검측가견시신경겸협+EGCG조겸협대측LGN적NF-L표체량고우시신경겸협+생리염수조(P=0.002);dLGN적nNOS양성세포계수재정상대조조、가수술+EGCG조、시신경겸협+EGCG조지간차이무통계학의의(P>0.05),이시신경겸협+생리염수조겸협대측dLGN적nNOS양성세포고우시신경겸협+EGCG조(P=0.000).결론 EGCG대대서시신경겸협상후LGN적신경원가능구유일정적보호작용,저충보호작용가능여EGCG억제료nNOS적표체유관.
Background Researches demonstrated that epigallocatechin-3-gallate(EGCG) can protect retinal ganglion cells(RGCs) against damage induced by retinal ischemia-reperfusion and optic nerve crush(ONC),but the effect of EGCG on lateral geniculate nucleus(LGN) was under study.Objective This study was designed to detect neuroprotective effect of EGCG on LGN in the rat model with ONC.Methods Forty-eight 7-week-old female clean Wistar rats were randomly divided into normal control group,sham operation+EGCG group,ONC+normal saline(NS) group and ONC+EGCG group.ONC models were created by clamping the optical nerve for 60 seconds with the clipper with the force of 40 grams in the right eyes of 24 rats.The EGCG solution(25mg/kg) was intraperitoneally injected from 2 days before operation daily for 5 consecutively days and orally administered(2mg/kg) after that,and NS was used in the same way for ONC+NS group.Four weeks after ONC,the brain tissue of the rats was obtained,and the neurons of dorsal LGN(dLGN) were counted by Nissl staining under the light microscopy.The expression of neurofilament triplet L(NF-L) was detected by immunohistochemistry and Western blot analysis.Meanwhile,the neuronal nitric oxide synthase(nNOS) positive cells were counted.Results Compared with normal control group,no significant differences were found in neuron number both between sham operation+EGCG group or ipsilateral LGN of operative eyes in ONC+normal saline group and ONC+EGCG group(P=0.906,0.561,0.794,0.646 respectively) in 4 weeks after ONC,but loss of neurons in contralateral LGN in both ONC+normal saline group and ONC+EGCG group were observed(P=0.000,0.015 respectively).However,compared with ONC+normal saline group,the density of neurons in ONC+EGCG group was higher(P=0.007).Moreover,a higher expression level of NF-L protein was seen in ONC+EGCG group compared with ONC+normal saline group at contralateral LGN of operative eyes(P=0.002).Concerning the number of nNOS positive cells in LGN,there was no significant difference among normal control group,sham operation+EGCG group and ONC+EGCG group(P＞0.05).The number of nNOS positive cells in the contralateral LGN of operative eyes of ONC+normal saline group was higher than that of ONC+EGCG group(P=0.000).Conclusion EGCG plays the protective effect on LGN after ONC in rats through mediating the expression of nNOS.
