榅桲总黄酮对高糖诱导的人晶状体上皮细胞氧化损伤的影响及作用机制  

Effects of total flavone of Cydonia oblonga on high glucose-induced oxidative damage in human lens epithelial cells and its mechanism

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作  者:罗元元[1] 曹静洁 王海营[1] 封传 唐陶富[1] 胡洁[1] LUO Yuanyuan;CAO Jingjie;WANG Haiying;FENG Chuan;TANG Taofu;HU Jie(School of Medical Technology,Yongzhou Vocational and Technical College,Yongzhou 425000,Hunan Province,China;Department of Ophthalmology,Xiangya Hospital of Central South University,Changsha 410005,Hunan Province,China)

机构地区:[1]湖南省永州职业技术学院医学技术学院,湖南省永州市425000 [2]中南大学湘雅医院眼科,湖南省长沙市410005

出  处:《眼科新进展》2024年第10期774-779,共6页Recent Advances in Ophthalmology

基  金:湖南省自然科学科教联合基金项目(编号:2019JJ70039)。

摘  要:目的探讨榅桲总黄酮对高糖诱导的人晶状体上皮细胞氧化损伤的影响及作用机制。方法用高糖诱导人晶状体上皮细胞建立细胞损伤模型。取人晶状体上皮细胞加入含有30 mmol·L^(-1)葡萄糖的培养基处理24 h,记为高糖组。用含有5.5 mmol·L^(-1)葡萄糖的培养基处理24 h记为对照组。取人晶状体上皮细胞按照每孔5×10^(3)个接种于96孔板,分别加入含有10 mmol·L^(-1)、20 mmol·L^(-1)、40 mmol·L^(-1)榅桲总黄酮与30 mmol·L^(-1)葡萄糖的培养基处理24 h,记为高糖+低榅桲总黄酮组、高糖+中榅桲总黄酮组、高糖+高榅桲总黄酮组。用Lipofectamine2000转染试剂向人晶状体上皮细胞分别转染anti-miR-NC、anti-miR-370,随后加30 mmol·L^(-1)葡萄糖处理24 h,命名为高糖+anti-miR-NC组、高糖+anti-miR-370组。分别向人晶状体上皮细胞转染miR-NC、miR-370 mimics,加30 mmol·L^(-1)葡萄糖的培养基及40 mmol·L^(-1)榅桲总黄酮处理24 h,记为高糖+榅桲总黄酮+miR-NC组、高糖+榅桲总黄酮+miR-370组。采用ELISA法监测超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量,流式细胞仪检测细胞凋亡率,qRT-PCR检测miR-370表达量,Western blot检测凋亡蛋白表达情况。结果与对照组相比,高糖组人晶状体上皮细胞SOD和CAT活性降低,MDA含量升高,差异均有统计学意义(均为P<0.05);与高糖组相比,高糖+低、中、高榅桲总黄酮组人晶状体上皮细胞SOD和CAT的活性升高,MDA含量降低,差异均有统计学意义(均为P<0.05)。与对照组相比,高糖组人晶状体上皮细胞凋亡率,Caspase-3、Caspase-9蛋白表达升高,差异均有统计学意义(均为P<0.05);与高糖组相比,高糖+低、中、高榅桲总黄酮组人晶状体上皮细胞凋亡率,Caspase-3、Caspase-9蛋白表达均降低,差异均有统计学意义(均为P<0.05)。对照组、高糖组、高糖+低榅桲总黄酮组、高糖+中榅桲总黄酮组、高糖+高榅桲总黄酮组人晶状�Objective To investigate the effect of total flavone of Cydonia oblonga on oxidative damage of human lens epithelial cells induced by high glucose and its mechanism.Methods A cell injury model was established by inducing human lens epithelial cells with high glucose.Human lens epithelial cells were cultured in the medium containing 30 mmol·L^(-1)glucose for 24 h,which was recorded as the high glucose group.Cells in the control group were cultured in a medium containing 5.5 mmol·L^(-1)glucose for 24 h.Human lens epithelial cells were inoculated into 96-well plates with 5×10^(3)cells per well,and treated with mediums containing 10 mmol·L^(-1),20 mmol·L^(-1),and 40 mmol·L^(-1)total flavone of Cydonia oblonga combined with 30 mmol·L^(-1)glucose for 24 h.They were recorded as high glucose+low total flavone of Cydonia oblonga group,high glucose+medium total flavone of Cydonia oblonga group,and high glucose+high total flavone of Cydonia oblonga group.Human lens epithelial cells were transfected with anti-miR-NC and anti-miR-370 with Lipofectamine2000 transfection reagent,and treated with 30 mmol·L^(-1)glucose for 24 h,which were recorded as high glucose+anti-miR-NC group and high glucose+anti-miR-370 group.Human lens epithelial cells were transfected with miR-NC and miR-370 mimics,and treated with medium containing 30 mmol·L^(-1)glucose and 40 mmol·L^(-1)total flavone of Cydonia oblonga for 24 h,which were labeled as high glucose+total flavone of Cydonia oblonga+miR-NC group and high glucose+total flavone of Cydonia oblonga+miR-370 group.The activities of superoxide dismutase(SOD)and catalase(CAT),and the content of malondialdehyde(MDA)were measured by Enzyme-linked immunosorbent assay;flow cytometry was applied to detect apoptosis rate;quantitative reverse transcription polymerase chain reaction was applied to detect the expression level of miR-370;Western blot was applied to detect the expression of apoptosis-related proteins.Results Compared with the control group,the activities of SOD and CAT decreased and

关 键 词:榅桲总黄酮 微小RNA-370 氧化应激 细胞凋亡 人晶状体上皮细胞 

分 类 号:R774.1[医药卫生—眼科]

 

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