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黄芪含药血清对氯化钴诱导ARPE-19细胞缺氧损伤的保护作用 被引量:4

Protective effect of astragalus-containing serum on cobalt chloride-induced hypoxia injury in ARPE-19 cells
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摘要 目的:研究黄芪含药血清对氯化钴(CoCl_(2))诱导人视网膜色素上皮(ARPE-19)细胞缺氧损伤的保护作用,从而探讨黄芪是否通过抗氧化应激来改善糖尿病视网膜病变(DR)。方法:建立CoCl_(2)诱导ARPE-19细胞缺氧模型,并分为以下5组:正常组(细胞正常培养,不加任何处理)、缺氧模型组(200μmol/L CoCl_(2))、空白血清组(200μmol/L CoCl_(2)+空白血清)、低剂量含药血清组(200μmol/L CoCl_(2)+10%含药血清)、高剂量含药血清组(200μmol/L CoCl_(2)+20%含药血清)。CCK-8检测细胞活性;试剂盒检测细胞上清液中还原型谷胱甘肽(GSH)、丙二醛(MDA)水平;ELISA检测细胞培养基中缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的含量;实时荧光定量PCR(qPCR)检测VEGF、HIF-1α和脯氨酰羟化酶-2(PHD-2)的mRNA水平;Western Blot法检测VEGF、HIF-1α和PHD-2的表达情况。结果:采用200μmol/L的CoCl_(2)浓度可成功建立ARPE-19细胞缺氧模型。低剂量和高剂量黄芪含药血清可抑制缺氧诱导的ARPE-19细胞增殖(P<0.05),升高ARPE-19细胞缺氧损伤中GSH水平,降低MDA含量(P<0.05)。低剂量和高剂量黄芪含药血清可抑制ARPE-19细胞缺氧损伤上清液中HIF-1α和VEGF的表达(P<0.05),以及ARPE-19细胞中VEGF、HIF-1α、PHD-2的mRNA表达和蛋白表达(P<0.05)。结论:低剂量和高剂量黄芪含药血清通过抗氧化作用减轻CoCl_(2)诱导ARPE-19细胞缺氧损伤。 AIM:To study the protective effect of astragalus-containing serum on cobalt chloride(CoCl_(2))-induced hypoxia injury of human retinal pigment epithelial cells(ARPE-19),so as to explore whether astragalus can improve diabetic retinopathy(DR)by anti-oxidative stress.METHODS:The ARPE-19 hypoxia model induced by CoCl_(2) was established and divided into the following 5 groups:normal group(cells were cultured normally without any treatment),hypoxia model group(200μmol/L CoCl_(2)),blank serum group(200μmol/L CoCl_(2)+blank serum),low-dose drug-containing serum group(200μmol/L CoCl_(2)+10%medicated serum)and high-dose drug-containing serum group(200μmol/L CoCl_(2)+20%medicated serum);CCK-8 detects cell viability;Detect the levels of reduced glutathione(GSH)and malondialdehyde(MDA)in the cell supernatant with a kit;ELISA was used to detect the content of hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)in cell culture medium;Real-time quantitative PCR(qPCR)to detect the mRNA levels of VEGF,HIF-1αand Prolyl hydroxylase-2(PHD-2);The expressions of VEGF,HIF-1αand PHD-2 were detected by Western Blot.RESULTS:Hypoxia model of ARPE-19 can successfully establish by CoCl_(2) at 200μmol/L.Low-dose and high-dose astragalus-containing serum could inhibit hypoxia-induced ARPE-19 proliferation(P<0.05),increase the GSH level and reduce the MDA content in ARPE-19 with hypoxic injury(P<0.05).Low-dose and high-dose astragalus-containing serum could inhibit the expression of HIF-1αand VEGF in ARPE-19 hypoxic injury supernatant(P<0.05),as well as the mRNA and protein expressions of VEGF,HIF-1αand PHD-2 in ARPE-19(P<0.05).CONCLUSION:Low-dose and high-dose astragalus-containing serum alleviates the hypoxia injury of ARPE-19 induced by CoCl_(2) through anti-oxidant effect.
作者 胡晗 王晓琴 聂浩 Han Hu;Xiao-Qin Wang;Hao Nie(Department of Ophthalmology,Jingzhou Hospital Affiliated to Yangtze University,Jingzhou 434020,Hubei Province,China;Medical School,Yangtze University,Jingzhou 434023,Hubei Province,China)
出处 《国际眼科杂志》 CAS 北大核心 2022年第6期899-903,共5页 International Eye Science
基金 国家自然科学基金资助项目(No.31502059) 湖北省教育厅科学研究计划指导性项目(No.B2015447)。
关键词 黄芪 糖尿病视网膜病变 氧化应激 血管内皮生长因子 缺氧诱导因子-1Α 人视网膜色素上皮(ARPE-19)细胞 astragalus diabetic retinopathy oxidative stress vascular endothelial growth factor hypoxia-inducible factor-1α human retinal pigment epithelial cells(ARPE-19)
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