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视网膜神经节细胞中AKAP1的丢失与青光眼小鼠病情发展的相关性及机制研究

Correlation between the loss of AKAP1 in retinal ganglion cells and the development of glaucoma mice and its mechanism
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摘要 目的探究视网膜神经节细胞中A激酶锚定蛋白1(AKAP1)的丢失与青光眼小鼠病情发展的相关性及机制。方法选择无特定病原体(SPF)级C57BL/6小鼠,通过硬膜外静脉阻塞并烧灼建立具有慢性高眼压的小鼠青光眼模型,然后按照眼压高低分为青光眼早期组(相对低眼压+造模后7 d)、青光眼中期组(相对中眼压+造模后15 d)和青光眼晚期组(相对高眼压+造模后28 d),每组6只,另选取6只正常小鼠作为对照组(造模后7 d)。手持眼压计检测各组小鼠眼内压;蛋白质印迹分析各组AKAP1的蛋白表达;苏木精-伊红染色用于分析视网膜的病理变化;CCK-8试剂盒与TUNEL分析检测细胞的活力与凋亡;免疫组织化学分析检测细胞自噬相关蛋白的表达水平;RT-PCR实时分析磷脂酰肌醇-3-激酶(PI3K)、丝苏氨酸蛋白激酶(Akt)和腺苷酸激活蛋白激酶(AMPK)的mRNA表达。结果与对照组相比,青光眼早期组、中期组和晚期组的AKAP1的蛋白表达、IPL和GCL厚度和细胞活力均显著降低,眼内压、细胞凋亡率、LC3-Ⅱ/Ⅰ和Beclin水平均显著升高(P <0.05),且随着病情发展,3组青光眼小鼠的AKAP1的蛋白表达、IPL和GCL厚度和细胞活力逐渐降低,眼内压、细胞凋亡率、LC3-Ⅱ/Ⅰ和Beclin水平均逐渐升高,组间差异均有统计学意义(P <0.05)。结论视网膜神经节细胞中AKAP1的丢失与青光眼小鼠病情发展负相关,AKAP1的低表达,抑制了PI3K/Akt/AMPK途径,促进了视网膜神经节细胞的凋亡和自噬,因此与青光眼的发病机制密切相关,有助于AKAP1针对青光眼的定向诊断和治疗。 Objective To explore the correlation and mechanism between the loss of AKAP1 in retinal ganglion cells and the development of glaucoma mice.Methods SPF grade C57BL/6 mice were selected to establish a mouse glaucoma model with chronic high intraocular pressure through epidural vein occlusion and cauterization,and according to the level of intraocular pressure they were divided into the early stage glaucoma group(relatively low intraocular pressure + 7 days after modeling),the mid-stage glaucoma group(relative middle intraocular pressure + 15 days after modeling) and late glaucoma group(relatively high intraocular pressure + 28 days after modeling),each group 6 mice,and another 6 normal mice were selected as the control group(7 days after modeling).Hand-held tonometer was used to detect intraocular pressure in each group of mice;Western blotting was used to analyze the protein expression of AKAP1 in each group;hematoxylin-eosin staining was used to analyze the pathological changes of the retina;CCK-8 kit and TUNEL was used to detect cell viability and apoptosis;Immunohistochemical analysis was used to detect the expression level of autophagy-related proteins;RT-PCR was used to analyze the mRNA expression of PI3K,Akt and AMPK in real time.Results Compared with the control group,the protein expression of AKAP1,the thickness of IPL and GCL and the cell viability of the early,middle and late glaucoma groups were significantly reduced,and the intraocular pressure,apoptosis rate,LC3-Ⅱ/Ⅰ and Beclin levels were significantly increased(P < 0.05).And with the development of the disease,the protein expression of AKAP1,the thickness of IPL and GCL and cell viability of the three groups of glaucoma mice gradually decreased,intraocular pressure,apoptosis rate,LC3-Ⅱ/Ⅰ and Beclin levels gradually increased,and the differences between groups were statistically significant(P < 0.05).Conclusion The loss of AKAP1 in retinal ganglion cells is negatively correlated with the development of glaucoma mice.The decreased expression of AKAP1 inhibits the PI3K/Akt/AMPK pathway and promotes apoptosis and autophagy of retinal ganglion cells,which is helpful for understanding of glaucoma Pathogenesis,and promote the targeted diagnosis and treatment of AKAP1 for glaucoma.
作者 沙金梅 董海莉 沙秀兰 SHA Jin-mei;DONG Hai-li;SHA Xiu-lan(Department of Ophthalmology,the Second Affiliated Hospital of Air Force Military Medical University,Xi'an Shaanxi 710038,China;Department of Stomatology,Baqiao Dongcheng Hospital of Xi'an,Xi'an Shaanxi 710000,China)
出处 《临床和实验医学杂志》 2021年第6期571-575,共5页 Journal of Clinical and Experimental Medicine
基金 陕西省科技攻关计划项目(编号:2018KS-14-03)。
关键词 小鼠 青光眼 视网膜神经节细胞 A激酶锚定蛋白1 PI3K/Akt/AMPK Mice Glaucoma Retinal ganglion cells A kinase anchor protein 1 PI3K/Akt/AMPK
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