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作 者:刘冬瑞[1] 宋凡倩 葛红岩[1] 郭嫱[1] 刘平[1] LIU Dong-rui;SONG Fan-qian;GE Hong-yan;GUO Qiang;LIU Ping(Department of Ophthalmology,the First Affiliated Hospital of Harbin Medical University,Harbin,Heilongjiang,150001,China;Qingdao Eye Hospital of Shandong First Medical University,Qingdao,Shandong,266071,China)
机构地区:[1]哈尔滨医科大学附属第一医院,黑龙江哈尔滨150001 [2]山东第一医科大学附属青岛眼科医院,山东青岛266071
出 处:《现代生物医学进展》2023年第16期3196-3200,共5页Progress in Modern Biomedicine
基 金:黑龙江省卫生健康委科研项目(2019-039)。
摘 要:目的:探索NDRG2基因在老年核性白内障晶状体组织中,随着核级增加的表达变化及可能的作用机制。方法:纳入2019年9月1日至2021年8月31日于哈尔滨医科大附属第一医院进行手术治疗的老年核性白内障患者的晶状体组织,依据Emery分级进行核分级,去除晶状体囊膜,收集晶状体裂解液,并依据蛋白浓度结果初步调齐各样本浓度。采用Western blot方法检测不同核分级晶状体NDRG2和Lamininγ2蛋白的表达差异。通过1200μM H_(2)O_(2)诱导HLE B-3晶状体上皮细胞96小时,诱导细胞衰老,构建老年性白内障的细胞模型;未经H2O2溶液诱导,培养96小时的晶状体上皮细胞作为对照组。收集H2O2诱导组及对照组细胞裂解液,Western blot方法检测两组NDRG2和Lamininγ2蛋白的表达差异。进一步构建pcDNA3.1-NDRG2/Lamininγ2质粒,转染入晶状体上皮细胞B3(lens epithelial cell,HLE B-3)细胞系,探索NDRG2参与到不同核级老年性白内障可能的作用机制。结果:老年核性白内障患者的晶状体中,NDRG2和Lamininγ2蛋白表达增加,随着核分级越高表达越多。H_(2)O_(2)处理的HLE B-3细胞衰老模型中,NDRG2和Lamininγ2蛋白表达较对照组增加。NDRG2过表达可以上调Lamininγ2蛋白表达,但Lamininγ2过表达对NDRG2蛋白无明显影响。结论:NDRG2在老年核性白内障晶状体组织中表达升高,并与核分级呈正相关,可能Lamininγ2蛋白参与了此过程的发展。Objective:To explore the expression level of NDRG2 protein in the lens tissues of the nuclear age related cataract patients and the possible mechanism.Methods:Collect lens without lens capsules from patients with nuclear age-related cataract undergoing surgical in First Affiliated Hospital,Harbin Medical University.Lens was grading though Emery method.And preliminarily adjust the concentration of each sample based on the protein concentration results.Detection of NDRG2 and Lamininγ2 expression level in different nuclear grading lenses by Western blot.HLE B-3 lens epithelial cell line was induced by 1200μM H_(2)O_(2)for 96h to establish the aging-realated cataract model.HLE B-3 cells without treated with H_(2)O_(2)were set as the control group.Collect lysates from the experimental group and the control group,and detect NDRG2 and Lamininγ2 in the two groups by Western blot.Further pCDNA3.1-NDRG2/Laminγ2 plasmids were transfected into HLE B-3 cells to explore the possible mechanism of NDRG2 involvement in different nuclear age related cataracts.Results:NDRG2 and Lamininγ2 were much higher in nuclear age-related cataract.The higher the nuclear grade,the more protein expression.In the H_(2)O_(2)treated HLE B-3 cell aging model,the expression level of NDRG2 and Lamininγ2 were much higher than the control group.NDRG2 overexpression couled up-regulate expression of Laminγ2,but Lamininγ2 overexpression had little influence to expression level of NDRG2.Conclusion:The expression of NDRG2 in nuclear age-related cataract lens tissues were increased and positively correlated with the nuclear grade,which might take effect by the upregulation of Lamininγ2.
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