[1]张新霞,王小敏,陆丽红,等.薯蓣皂苷对高糖诱导的视网膜色素上皮细胞损伤的保护作用[J].眼科新进展,2022,42(4):278-283.[doi:10.13389/j.cnki.rao.2022.0056]
　ZHANG Xinxia,WANG Xiaomin,LU Lihong,et al.Protective effect of diosgenin on retinal pigment epithelial cell injury induced by high glucose[J].Recent Advances in Ophthalmology,2022,42(4):278-283.[doi:10.13389/j.cnki.rao.2022.0056]
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薯蓣皂苷对高糖诱导的视网膜色素上皮细胞损伤的保护作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
42卷
期数:
2022年4期
页码:
278-283
栏目:
实验研究
出版日期:
2022-04-05

文章信息/Info

Title:
Protective effect of diosgenin on retinal pigment epithelial cell injury induced by high glucose
作者:
张新霞; 王小敏; 陆丽红; 王保君; 狄文玉
453100　河南省新乡市，新乡医学院第一附属医院眼科（张新霞，王小敏，陆丽红，王保君）；453100　河南省新乡市，新乡医学院第一附属医院病理科（狄文玉）
Author(s):
ZHANG Xinxia1; WANG Xiaomin1; LU Lihong1; WANG Baojun1; DI Wenyu2
1.Department of Ophthalmology,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,Henan Province,China
2.Department of Pathology,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,Henan Province,China
关键词:
薯蓣皂苷; 糖尿病视网膜病变; 视网膜色素上皮细胞; 高糖; 氧化应激; 细胞凋亡
Keywords:
diosgenin;  diabetic retinopathy;  retinal pigment epithelium cells;  high glucose;  oxidative stress;  apoptosis
分类号:
R966;R774.1
DOI:
10.13389/j.cnki.rao.2022.0056
文献标志码:
A
摘要:
目的　探讨薯蓣皂苷（Dio）对高糖（HG）诱导的视网膜色素上皮（RPE）细胞损伤的保护作用，并分析其机制。方法　用细胞计数试剂8（CCK-8）法筛选葡萄糖浓度和无细胞毒性的Dio剂量范围。将ARPE-19细胞分为对照组（5 mmol·L-1葡萄糖处理48 h）、模型组（50 mmol·L-1葡萄糖处理48 h）和低、中、高剂量Dio处理组（分别用50 mmol·L-1葡萄糖联合0.5 mmol·L-1 、2.0 mmol·L-1 和8.0 mmol·L-1 Dio处理48 h）。CCK-8法检测细胞活性；FITC标记膜联蛋白-V/碘化丙啶（Annexin V-FITC/PI）染色检测细胞凋亡；荧光探针法检测活性氧（ROS）水平；试剂盒法检测丙二醛（MDA）水平、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）活性；JC-1染色检测线粒体膜电位；Western blot法检测B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、cleaved Caspase-3和Yes相关蛋白（YAP）的表达水平；免疫荧光法观察YAP的表达。结果　筛选的葡萄糖浓度为50 mmol·L-1，Dio的无细胞毒性的剂量为0.5 mmol·L-1、2.0 mmol·L-1、8.0 mmol·L-1。与对照组比较，模型组细胞活性、线粒体膜电位、CAT和GSH-Px活性以及Bcl-2蛋白和YAP蛋白表达水平均明显降低（均为P<0.001），细胞凋亡水平、ROS和MDA水平以及cleaved Caspase-3蛋白和Bax蛋白表达水平均明显升高（均为P<0.001）；与模型组比较，低、中、高剂量Dio处理组细胞活性、线粒体膜电位、CAT和GSH-Px活性以及Bcl-2蛋白和YAP蛋白表达水平均明显升高(均为P<0.05)，细胞凋亡水平、ROS和MDA水平以及cleaved Caspase-3蛋白和Bax蛋白表达水平均明显降低，且均呈剂量依赖性（均为P<0.05)。结论　Dio可通过激活YAP信号，降低氧化应激反应和抑制线粒体介导的细胞凋亡途径，进而减轻HG诱导的RPE细胞损伤。
Abstract:
Objective　To investigate the protective effect of diosgenin (Dio) on retinal pigment epithelial (RPE) cell injury induced by high glucose (HG), and to analyze its mechanism. Methods　Glucose concentration and non-cytotoxic Dio dose range were screened by CCK-8 assay. ARPE-19 cells were divided into the control group (treated with 5 mmol·L-1 glucose for 48 h), model group (treated with 50 mmol·L-1 glucose for 48 h), and low-, medium- and high-dose Dio groups (treated with 50 mmol·L-1 glucose combined with 0.5 mmol·L-1, 2.0 mmol·L-1 and 8.0 mmol·L-1 Dio, respectively for 48 h). Cell viability was detected by CCK-8 assay. Apoptosis was detected by Annexin V-FITC/PI staining. The levels of reactive oxygen species (ROS) were measured by fluorescence probe. The level of malondialdehyde (MDA) and the activity of catalase (CAT) and glutathione peroxidase (GSH-Px) were measured by assay kits. Mitochondrial membrane potential was tested by JC-1 staining. The expression levels of B-cell lymphoma/leukaemia-2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved Caspase-3 and Yes-associated protein (YAP) were tested by Western blot. The expression of YAP was observed by immunofluorescence.Results　The screened glucose concentration was 50 mmol·L-1, and the non-cytotoxic dose range of Dio was 0.5 mmol·L-1, 2.0 mmol·L-1 and 8.0 mmol·L-1. Compared with the control group, cell viability, mitochondrial membrane potential, CAT and GSH-Px activity, Bcl-2 and YAP levels in the model group were significantly decreased (all P<0.001), while apoptosis, ROS and MDA levels, cleaved Caspase-3 and Bax levels were significantly increased (all P<0.001). Compared with the model group, cell viability, mitochondrial membrane potential, CAT and GSH-Px activity, Bcl-2 and YAP levels in low-, medium- and high-dose Dio groups were significantly increased, while apoptosis, ROS and MDA levels, cleaved Caspase-3 and Bax levels were significantly decreased, all in a dose-dependent manner(all P<0.05).Conclusion　Dio can reduce HG-induced RPE cell injury by activating YAP signal, reducing oxidative stress response and inhibiting mitochondria-mediated apoptosis pathway.

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备注/Memo

备注/Memo:
河南省医学科技攻关计划项目（编号：SBGJ2018055）
更新日期/Last Update: 2022-04-05