[1]刘文强,冯闯,左中夫,等.PI3K/Akt信号通路对高糖状态下视网膜Müller细胞的影响[J].眼科新进展,2020,40(11):1024-1028.[doi:10.13389/j.cnki.rao.2020.0229]
 LIU Wenqiang,FENG Chuang,ZUO Zhongfu,et al.Effect of PI3K/Akt signaling pathway on retinal Müller cells in diabetic rats[J].Recent Advances in Ophthalmology,2020,40(11):1024-1028.[doi:10.13389/j.cnki.rao.2020.0229]
点击复制

PI3K/Akt信号通路对高糖状态下视网膜Müller细胞的影响/HTML
分享到:

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年11期
页码:
1024-1028
栏目:
实验研究
出版日期:
2020-11-05

文章信息/Info

Title:
Effect of PI3K/Akt signaling pathway on retinal Müller cells in diabetic rats
作者:
刘文强冯闯左中夫刘学政
121001 辽宁省锦州市,锦州医科大学基础医学院解剖学教研室(刘文强,左中夫,刘学政);110847 辽宁省沈阳市,辽宁中医药大学研究生学院(冯闯)
Author(s):
LIU Wenqiang1FENG Chuang2ZUO Zhongfu1LIU Xuezheng1
1.Department of Anatomy,School of Basic Medicine,Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China
2.Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning Province,China
关键词:
糖尿病Müller细胞PI3K/Akt信号通路凋亡
Keywords:
diabetes Müller cells PI3K/Akt signaling pathway apoptosis
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2020.0229
文献标志码:
A
摘要:
目的 探讨PI3K/Akt信号通路对高糖状态下视网膜Müller细胞的影响及机制。方法 本实验分两部分,动物实验:雄性SD大鼠随机分成对照组、糖尿病组、胰岛素样生长因子-1(insulin-like growth factors-1,IGF-1)组、IGF-1+LY294002(PI3K/Akt信号通路抑制剂)组。后三组采用链脲佐菌素(streptozotocin,STZ)(50 mg·kg-1)诱导成为糖尿病模型。模型诱导成功后,IGF-1组给予IGF-1 (80 g·L-1) 5 μL玻璃体内注射给药,IGF-1+LY294002组给予5 μL(IGF-1+ LY294002)(80 μg·L-1)玻璃体内注射给药。细胞实验:对培养的Müller细胞进行不同处理,分为对照组(空白对照)、高糖组(30 mmol·L-1葡萄糖)、IGF-1组(30 mmol·L-1葡萄糖+80 μg·L-1 IGF-1)、IGF-1+LY294002组[30 mmol·L-1葡萄糖+80 μg·L-1(IGF-1+ LY294002)]。HE染色检测大鼠视网膜内核层(inner nuclear layer,INL)细胞密度,免疫荧光染色检测大鼠视网膜神经胶质纤维酸性蛋白(glial fibrillary acid protein,GFAP)表达及Müller细胞Caspase-3表达,TUNEL染色检测细胞凋亡情况,Western blot检测p-Akt、NF-κB、Caspase-3蛋白相对表达水平。结果 动物实验中,与对照组相比,糖尿病组及IGF-1+LY294002组GFAP表达均明显增加,INL细胞密度均明显降低(均为P<0.05);与糖尿病组相比,IGF-1组GFAP表达明显下降,INL细胞密度明显增加(均为P<0.05)。细胞实验中,与对照组相比,高糖组及IGF-1+LY294002组细胞凋亡率和NF-κB、Caspase-3蛋白相对表达水平均明显增加,p-Akt蛋白相对表达均明显降低(均为P<0.05);而与高糖组相比,IGF-1组细胞凋亡率、NF-κB及Caspase-3蛋白相对表达均明显下降,p-Akt蛋白相对表达明显增加(均为P<0.05)。结论 激活PI3K/Akt通路可对抗高糖状态下视网膜Müller细胞凋亡,其机制可能与下调NF-κB、Caspase-3蛋白表达有关。
Abstract:
Objective To investigate the effect and mechanism of PI3K/Akt signaling pathway on retinal Müller cells in diabetic rats.Methods The experiment was divided into two parts. As for animal experiment,male SD rats were randomly divided into control group, diabetic group, insulin-like growth factors (IGF-1) group and IGF-1+ LY294002 (PI3K/Akt signaling pathway inhibitor) group. Diabetes model induced by streptozotocin(STZ) (50 mg·kg-1) in the latter three groups. After successful induction of the model, the IGF-1 group was given intravitreal injection of 5 μL IGF-1 (80 μg·L-1), and the IGF-1+LY294002 group was given 5 μL (IGF-1+ LY294002) (80 μg·L-1). As for cell experiment, the cultured Müller cells were divided into control group (blank control), high glucose group (30 mmol·L-1 glucose), IGF-1 group (30 mmol·L-1 glucose+80 μg·L-1 IGF-1), IGF-1+LY292 group [30 mmol·L-1 glucose+80 μg·L-1 (IGF-1 + LY294002)]. HE staining was used to detect the density of retinal inner nuclear layer (INL) cells, immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP)in rat retina and the expression of Caspase-3 in Müller cells, TUNEL staining was used to detect apoptosis of Müller cells, and Western blot was used to detect the relative expression of p-Akt, NF-κB and Caspase-3 proteins.Results In the animal experiment, when compared with the control group, the expression level of GFAP in diabetes group and IGF-1+LY294002 group increased significantly, and the density of INL cells decreased significantly(both P<0.05). Compared with diabetes group, the expression level of GFAP in IGF-1 group decreased significantly, and the density of INL cells increased significantly(both P<0.05). In the cell experiment,when compared with the control group, the apoptotic rate and the expression levels of NF-κB and Caspase-3 in high glucose group and IGF-1+LY294002 group increased significantly, while the expression level of p-Akt decreased significantly (both P<0.05); compared with high glucose group, the apoptotic rate and the expression level of NF-κB and Caspase-3 in IGF-1 group decreased significantly, and the expression level of p-Akt increased significantly (all P<0.05).Conclusion Activation of PI3K/Akt pathway can inhibit diabetic retinal Müller cells apoptosis, which may be related to down-regulation the protein expression of NF-κB and Caspase-3.

参考文献/References:

[1] WANG J,O’SULLIVAN M L,MUKHERJEE D,PUAL V M,FARSIU S,KAY J N.Anatomy and spatial organization of müller glia in mouse retina[J].J Comp Neurol,2017,525(8):1759-1777.
[2] QIU A W,BIAN Z,MAO P A,LIU Q H.IL-17A exacerbates diabetic retinopathy by impairing Müller cell function via Act1 signaling[J].Exp Mol Med,2016,48(12):e280.
[3] SANG Q,SUN D,CHEN Z,ZHAO W.NGF and PI3K/Akt signaling participate in the ventral motor neuronal protection of curcumin in sciatic nerve injury rat models[J].Biomed Pharmacother,2018,103(7):1146-1153.
[4] JIANG G,WANG C,ZHANG J,LIU H.Mediation of insulin growth factor-1 in Alzheimers disease and the mechanism of PRNP genetic expression and the PI3K/Akt signaling pathway[J].Exp Ther Med,2017,13(6):2763-2766.
[5] HU J,GRUBER K J,LIU H,ZHAO H,GARCIA A A.Diabetes knowledge among older adults with diabetes in Beijing,China[J].J Clin Nurs,2013,22(1-2):51-60.
[6] SORRENTINO F S,MATTEINI S,BONIFAZZI C,SEBASTIANI A,PARMEGGIANI F.Diabetic retinopathy and endothelin system:microangiopathy versus endothelial dysfunction[J].Eye,2018,32(7):1157-1163.
[7] KHAN M A,KATTA M,GURUNADH V S,MOULICK P S,GUPTA S,SATI A.A clinical correlation of conjunctival microangiopathy with grades of retinopathy in type 2 diabetes mellitus[J].Med J Armed Forces India,2017,73(3):261-266.
[8] KIRSCH M,TRAUTMANN N,ERNST M,HOFMANN H D.Involvement of gp130-associated cytokine signaling in müller cell activation following optic nerve lesion[J].Glia,2010,58(7):768-779.
[9] BARBER A J,LIETH E,KHIN S A,ANTONETTI D A,BUCHANAN A G,GARDNER T W.Neural apoptosis in the retina during experimental and human diabetes.Early onset and effect of insulin[J].J Clin Invest,2018,102(4):783-791.
[10] 梁汇珉,刘学政.脑源性神经营养因子(BDNF)对高糖环境下视网膜Müller细胞的保护作用[J].眼科新进展,2018,38(8):714-718.
LIANG H M,LIU X Z.Effect of brain-derived neurotrophic factor on high glucose induced apoptosis of retinal Müller cell[J].Rec Adv Ophthalmol,2018,38(8):714-718.
[11] HUANG C,WEN C,YANG M,GAN D,FAN C,LI A,et al.Lycopene protects against t-BHP-induced neuronal oxidative damage and apoptosis via activation of the PI3K/Akt pathway[J].Mol Biol Rep,2019,46(3):3387-3397.
[12] 杨帆,周其全.PI3K/Akt信号通路与神经保护[J].国际脑血管病杂志,2013,21(2):143-147.
YANG F,ZHOU Q Q.PI3K/Akt signaling pathway and neuroprotection[J].Int J Cer Dis,2013,21(2):143-147.
[13] 邓菲,李艳,尹珊,贾凡,栾莉.PEDF对高糖刺激下大鼠视网膜Müller细胞NF-κB和Caspase-3表达的影响[J].潍坊医学院学报,2019,41(2):50-53,59.
DENG F,LI Y,YIN S,JIA F,LUAN L.Effect of PEDF on the expression of NF -κB and Caspase-3 on high glucose-stimulated rat retinal Müller cells[J].J Weifang Med Coll,2019,41(2):50-53,59.
[14] PAN Y,YE C,TIAN Q,YAN S,ZENG X,XIAO C,et al.miR-145 suppresses the proliferation,invasion and migration of NSCLC cells by regulating the BAX/BCL-2 ratio and the caspase-3 cascade[J].Oncol Lett,2018,15(4):4337-4343.
[15] ALI D,TRIPATHI A,AL ALI H,SHAHI Y,MISHRA K K,ALARIFI S,et al.ROS-dependent Bax/Bcl2 and caspase 3 pathway-mediated apoptosis induced by zineb in human keratinocyte cells[J].Oncotargets Ther,2018,11(1):489-497.

相似文献/References:

[1]丁文君 韦琦 梁皓 陈金卯 李霞 谭少健.糖尿病兔后发性白内障发生过程中晶状体上皮细胞增殖的变化[J].眼科新进展,2012,32(1):000.
[2]陈放 徐珊 吕伟红 程宏 张洪泉.糖尿病大鼠视网膜的氧化应激损伤及葛根素对其的干预作用[J].眼科新进展,2012,32(1):000.
[3]李晓艳 张卯年.褪黑素对糖尿病大鼠早期视网膜神经细胞凋亡的影响[J].眼科新进展,2012,32(6):000.
[4]楚静 陈忠平.普拉洛芬及羟苯磺酸钙在糖尿病患者白内障手术中的应用[J].眼科新进展,2013,33(8):000.
[5]张曙光 何跃 项杰 马林昆 李燕 袁援生.JNK2在早期糖尿病小鼠视网膜中的表达及作用[J].眼科新进展,2012,32(4):000.
[6]李艳 潘斌 李筱荣 袁佳琴 张杰.糖尿病大鼠视网膜PEDF表达与神经退行性改变[J].眼科新进展,2012,32(8):000.
[7]崔颖 郭海科 韩云飞 孟倩丽 张良 尹东明.2型糖尿病住院患者糖尿病视网膜病变患病率及危险因素分析[J].眼科新进展,2012,32(8):000.
[8]张谊 宋光辉 杨架林 张钰 肖强 潘璐 赵伟红 刘昊.糖尿病患者短期内血糖变化对黄斑区视网膜厚度的影响[J].眼科新进展,2012,32(10):000.
[9]赵恳.白内障超声乳化人工晶状体植入术对糖尿病患者角膜内皮细胞形态与功能的影响[J].眼科新进展,2013,33(4):000.
[10]任浩宇 胡凯耀 杨忠.视网膜再生的细胞来源与Müller细胞去分化机制[J].眼科新进展,2013,33(5):000.

备注/Memo

备注/Memo:
国家自然科学基金资助(编号:81571383);中国博士后科学基金资助(编号:2017M612870)
更新日期/Last Update: 2020-11-05