[1]向艳,高晶,罗班,等.慢性高眼压模型鼠视网膜神经节细胞及视神经中9位丝氨酸磷酸化糖原合酶激酶3β的表达变化[J].眼科新进展,2017,37(4):326-329.[doi:10.13389/j.cnki.rao.2017.0082]
　XIANG Yan,GAO Jing,LUO Ban,et al.Expression changes of phosphorylation glogyen synthase kinase 3β at ser9 in retinal ganglion cell and optic nerve of rat ocular hypertension model[J].Recent Advances in Ophthalmology,2017,37(4):326-329.[doi:10.13389/j.cnki.rao.2017.0082]
点击复制

慢性高眼压模型鼠视网膜神经节细胞及视神经中9位丝氨酸磷酸化糖原合酶激酶3β的表达变化/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年4期
页码:
326-329
栏目:
实验研究
出版日期:
2017-04-05

文章信息/Info

Title:
Expression changes of phosphorylation glogyen synthase kinase 3β at ser9 in retinal ganglion cell and optic nerve of rat ocular hypertension model
作者:
向艳; 高晶; 罗班; 陈博; 李贵刚; 王玮; 张虹; 王平
430030　湖北省武汉市，华中科技大学同济医学院附属同济医院眼科
Author(s):
XIANG Yan; GAO Jing; LUO Ban; CHEN Bo; LI Gui -Gang; WANG Wei; ZHANG Hong; WANG Ping
Department of Ophthalmology，Tongji Hospital，Tongji Medical College，Huazhong University of Science and Technology，Wuhan 430030，Hubei Province，China
关键词:
慢性高眼压模型鼠; 糖原合酶激酶3β; 磷酸化; 视网膜神经节细胞; 视神经
Keywords:
ocular hypertension model rat; glogyen synthase kinase 3β; phosphorylation; retinal ganglion cell; optic nerve
分类号:
R774.6
DOI:
10.13389/j.cnki.rao.2017.0082
文献标志码:
A
摘要:
目的　观察慢性高眼压模型鼠视网膜神经节细胞（retina ganglion cells，RGCs）和视神经中9位丝氨酸磷酸化糖原合酶激酶3β［p-GSK3β（ser9）］的表达变化，探讨GSK3β是否在慢性高眼压RGCs及视神经退行性病变中发挥作用。方法　取健康SD大鼠30只，随机分为3组：模型对照组、慢性高眼压模型2周组、慢性高眼压模型4周组，每组10只。取大鼠右眼，采用巩膜上静脉结扎并烧灼法建立大鼠慢性高眼压模型。分别于造模后2周和4周取5只大鼠眼球行冰冻切片，尼氏染色观察RGCs数目，免疫荧光染色观察RGCs和视神经中p-GSK3β(ser9)的变化；各组取另外5只大鼠眼视网膜，Western blot检测p-GSK3β(ser9)及总-GSK3β的表达。结果　造模前3组大鼠右眼眼压差异无统计学意义（P=0.89）；造模后3组间眼压差异有统计学意义（P<0.01），慢性高眼压模型2周组和4周组眼压与模型对照组比较明显升高，分别升高了59.13%和26.93%，差异均有统计学意义 (均为 P<0.01）。视网膜尼氏染色RGCs计数发现模型对照组RGCs数为(149±12)个，慢性高眼压模型2周组和4周组RGCs数较模型对照组明显减少，分别为(120±10)个和(86±7)个，差异均有统计学意义（均为 P<0.05）；且4周组比2周组进一步减少（P<0.01）。慢性高眼压模型2周组和4周组视网膜中p-GSK3β(ser9）阳性着色，同模型对照组相比，其RGCs层及视神经中p-GSK3β(ser9）染色荧光减弱；4周组较模型对照组减弱更明显。与模型对照组相比，慢性高眼压模型2周组和4周组视网膜中总-GSK3β的表达差异无统计学意义（F=0.24，P=0.79）；而3组间p-GSK3β(ser9)表达差异有统计学意义（P<0.01），与模型对照组相比，慢性高眼压模型2周组和4周组视网膜中p-GSK3β(ser9)分别减少了19.89%和36.46%（均为P<0.05）。慢性高眼压模型4周组比2周组视网膜中p-GSK3β(ser9)表达持续减少，差异具有统计学意义（P<0.05）。结论　慢性高眼压模型鼠RGCs数目减少，RGCs和视神经中p-GSK3β(ser9)表达减少，推测p-GSK3β(ser9)表达变化可能参与了慢性高眼压模型鼠RGCs及视神经退行性病变。
Abstract:
Objective　To observe the expression change of phosphorylation glogyen synthase kinase 3β at Ser9 ［p-GSK3β (Ser9)］ and investigate whether GSK3β involved in the retinal ganglion cell (RGC) and optic nerve in rat ocular hypertension model.Methods　Thirty health Sprague Dawley (SD) rats with normal intraocular pressure (IOP) were randomly divided into 3 groups（10 cases in each group）:control group,2 weeks ocular hypertension(OHT) group and 4 weeks OHT group.The ocular hypertension model was established by using episcleral veins ligation and cauterization.5 eyes from each group were taken for frozen section at week 2 and week 4 after establishing ocular hypertension model.The number of RGC was counted with Nissl’s staining.The expression and distribution of p-GSK3β (Ser9) in RGC and optic nerve were detected by immunofluorescence staining.The retinal sample of other 5 rats was harvested for detecting the expression of total glogyen synthase kinase 3β (total GSK3β) and p-GSK3β (Ser9) by Western blot.Results　There was no statistical difference in IOP before modeling among three groups (P=0.89),but there was statistical difference after modeling (P<0.01),compared with the control group,the increased rate of IOP in 2 weeks OHT group and 4 weeks OHT group were 59.13% and 26.93% (all P<0.05).The average RGC numbers of 2 weeks OHT group and 4 weeks OHT group were 120±10 per eye and 86±7 per eye,which were both less than the number 149±12 per eye in control rats，there were statistical differences (all P<0.05).Furthermore,the number of RGC in 4 weeks OHT group was less than 2 weeks OHT group (P<0.01).The expression of p-GSK3β (Ser9) in the retina and optic nerve were positive detected by immunofluorescence,and the intensity of p-GSK3β (Ser9) immunofluorescence staining in RGC and optic nerve of OHT rats were less than model control eyes.The amount of p-GSK3β (Ser9) in retina of 2 weeks OHT rats decreased 19.89% compared with control group rats（P<0.05），and the amount of p-GSK3β (Ser9) in retina of 4 weeks OHT rats was significantly decreased 36.46% compared with control group rats（all P<0.05）.While the total GSK3β did not change（P>0.05）.Conclusion　The number of RGC and the expression of p-GSK3β (Ser9) in RGC and optic nerve of OHT rat eyes decreased significantly.The change expression of the p-GSK3β (Ser9) in the RGCs and optic nerve may be correlated with RGC neurodegeneration in glaucomatous disorders.

参考文献/References:

［1］　NICKELLS RW.From ocular hypertension to ganglion cell death a theoretical sequence of events leading to glaucoma［J］.Can J Ophthalmol，2007，42 (2)：278-287.
［2］　KOH SH，KIM Y，KIM HY，HWANG S，LEE CH，KIM SH.Inhibition of glycogen synthase kinase-3 suppresses the onset of symptoms and disease progression of G93 A-SOD1 mouse model of ALS［J］.Exp Neurol，2007，205(2)：336-346.
［3］　GARCA-GOROSTIAGA I，SNNCHEZ-JUAN P，MATEO I，RODRíGUEZ-RODRGUEZ E，SNCHEZ-QUINTANA C，DEL OLMO SC，et al.Glycogen synthase kinase-3 beta and tau genes interact in Parkinson’s and Alzheimer’s diseases［J］.Ann Neurol，2009，65(6)：759-761.
［4］　CARMICHAEL J，SUGARS KL，BAO YP，RUBINSZTEIN DC.Glycogen synthase kinase-3β inhibitors prevent cellular polyglutamine toxicity caused by the Huntington’s disease mutation［J］.J Biol Chem，2002，277(37)：33791-33798.
［5］　JANSSEN SF，JANSONIUS NM，BOUWMAN F，VERBRAAK FD，BERGEN AA.Systematic review of the association between Alzheimer’s disease and chronic glaucoma［J］.Clin Ophthalmol，2015，4(9)：783-784.
［6］　ERASLAN M，ERMAN E，EKI O，BALCI S，DERICIOGˇLU V，SAHIN ，et al.Neurodegeneration in ocular and central nervous systems：optical coherence tomography study in normal-tension glaucoma and Alzheimer disease［J］.Turk J Med Sci，2015，45(5)：1106-1114.
［7］　CHIASSEU M，CUEVARGAS JL，DESTROISMAISONS L，VANDE VELDE C，LECLERC N，DIPOLO A.Tau accumulation，altered phosphorylation，and missorting promote neurodegeneration in glaucoma［J］.J Neurosci，2016，36(21)：5785-5798.
［8］　RUSSO R，CAVALIERE F，BERLIOCCHI L，NUCCI C，GLIOZZI M，MAZZEI C，et al.Modulation of pro-survival and death associated pathways under retinal ischemia/reperfusion：effects of NMDA receptor blockade［J］.J Neurochem，2008，107(5)：1347-1357.
［9］　石慧，向艳，陈岚，陈志祺，李贵刚，张虹.氯化锂治疗慢性青光眼神经退行性病变的作用机制［J］.眼科新进展，2013，33(10)：922-926.
SHI H，XIANG Y，CHENG L，CHEN ZQ，LI GG，ZHANG H.The effective mechanism of Lithium chloride on treatment of chronic glaucoma about neural degenerative diseases［J］.Rec Adv Ophthalmol，2013，33(10)：922-926.
［10］　PEINEAU S，BRADLEY C，TAGHIBIGLOU C，DOHERTY A，BORTOLOTTO ZA，WANG YT，et al.The role of GSK-3 in synaptic plasticity［J］.Br J Pharmacol，2008，153(Suppl 1)：S428-437.
［11］　LONG ZM，ZHAO L，JIANG R，WANG KJ，LUO SF，ZHENG M，et al.Valproic acid modifies synaptic structure and accelerates neurite outgrowth via the glycogen synthase kinase-3β signaling pathway in an Alzheimer’s disease model［J］.CNS Neurosci Ther，2015，21(11)：887-897.
［12］　D’ANGELO B，EK CJ，SUN Y，ZHU C，SANDBERG M，MALLARD C.GSK3β inhibition protects the immature brain from hypoxic-ischaemic insult via reduced STAT3 signalling［J］.Neuropharmacology，2016，101：13-23.
［13］　KING TD，JOPE RS.Inhibition of GSK3 protects cells from intrinsic but not extrinsic oxidative stress［J］.Neuroreport，2005，16(6)：597-601.
［14］　CROWDER RJ，FREEMAN RS.Glycogen synthase kinase-3β activity is critical for neuronal death caused by inhibiting PI 3-kinase or Akt but not for death caused by NGF withdrawal［J］.J Biol Chem，2000，275(44)：34266 -34271.
［15］　PAN YY，DENG Y，XIE S，WANG ZH，WANG Y，REN J，et al.Altered wnt signaling pathway in cognitive impairment caused by chronic intermittent hypoxia：focus on glycogen synthase kinase-3β and β-catenin［J］.Chin Med J (Engl)，2016，129(7)：838-845.
［16］　BHOWMIK M，KHANAM R，SAINI N，VOHORA D.Activation of AKT/GSK3β pathway by TDZD-8 attenuates kainic acid induced neurodegeneration but not seizures in mice［J］.Neurotoxicology，2015，46：44-52.
［17］　JIN N，YIN X，YU D，CAO M，GONG CX，IQBAL K，et al.Truncation and activation of GSK3β by calpain I：a molecular mechanism links to tau hyperphosphorylation in Alzheimer’s disease［J］.Sci Rep，2015，5：8187.
［18］　CHEN Y，SU Y，RUN X，SUN Z，WANG T，SUN S，et al.Pretreatment of PC12 cells with 17β-estradiol prevents Aβ-induced down-regulation of CREB phosphorylation and prolongs inhibition of GSK3β［J］.J Mol Neurosci，2013，50(3)：394-401.
［19］　LIN CF，TSAI CC，HUANG WC，WANG YC，TSENG PC，TSAI TT，et al.Glycogen synthase kinase-3β and caspase-3 mediate ceramide- and etoposide-induced apoptosis by regulating the lysosomal-mitochondrial axis［J］.PLoS One，2016，11(1)：e0145460-0145473.
［20］　JEON GS，KIM JE，AHN SW，PARK KS，HONG YH，YE IH，et al.Effect of JGK-263 as a new glycogen synthase kinase-3β inhibitor on extrinsic apoptosis pathway in motor neuronal cells［J］.Biochem Biophys Res Commun，2013，439(2)：309-314.
［21］　MAQBOOL M，MOBASHIR M，HODA N.Pivotal role of glycogen synthase kinase-3：A therapeutic target for Alzheimer’s disease［J］.Eur J Med Chem，2016，107：63-81.
［22］　SACC SC，GANDOLFI S，BAGNIS A，MANNI G，DAMONTE G，TRAVERSO CE，et al.From DNA damage to functional changes of the trabecular meshwork in aging and glaucoma［J］.Ageing Res Rev，2016，29：26-41.
［23］　TANITO M，KAIDZU S，TAKAI Y，OHIRA A.Association between systemic oxidative stress and visual field damage in open-angle glaucoma［J］.Sci Rep，2016，6：25792.

备注/Memo

备注/Memo:
湖北省自然科学基金资助(编号：2014CFB442)
更新日期/Last Update: 2017-04-26