目的 探讨莫诺苷(Morroniside,Mor)对高糖诱导的人视网膜血管内皮细胞(human retinal microvascular endothelial cells,HRECs)氧化应激、凋亡的影响及其分子机制。 方法 将HRECs分为对照组(Con)、高糖组(HG)、HG+Mor-低剂量组(L)、HG+Mor-中剂量组(M)、HG+Mor-高剂量组(H)、HG+anti-miR-NC组、HG+anti-miR-483-5p组、HG+Mor+miR-NC组、HG+Mor+miR-483-5p组。流式细胞仪检测HRECs凋亡率及ROS水平,酶联免疫吸附检测MDA水平和SOD活性。实时定量PCR检测miR-483-5p表达。 结果 与Con组比较,HG组HRECs凋亡率、ROS和MDA水平升高(P<0.05),SOD活性降低(P<0.05),miR-483-5p表达升高(P<0.05)。与HG组比较,HG+Mor-L组、HG+Mor-M组、HG+Mor-H组HRECs凋亡率、ROS和MDA水平降低(P<0.05),SOD活性升高(P<0.05),miR-483-5p表达降低(P<0.05)。与HG+anti-miR-NC组比较,HG+anti-miR-483-5p组HRECs凋亡率、ROS和MDA水平降低(P<0.05),SOD活性升高(P<0.05)。与HG+Mor+miR-NC组比较,HG+Mor +miR-483-5p组HRECs凋亡率、ROS和MDA水平升高(P<0.05),SOD活性降低(P<0.05)。 结论 莫诺苷通过下调miR-483-5p表达可抑制高糖诱导的HRECs氧化应激和凋亡。
Abstract
Objective To investigate the effect and molecular mechanism of Morroniside (Mor) on high glucose-induced human retinal microvascular endothelial cells (HRECs) oxidative stress and apoptosis. Methods HRECs were divided into a control (Con) group, a high glucose (HG) group, a HG+Mor-low dose (L) group, a HG+Mor-medium dose (M) group, a HG+Mor-high dose (H) Group, a HG+anti-miR-NC group, a HG+anti-miR-483-5p group, a HG+Mor+miR-NC group, a HG+Mor+miR-483-5p group. The apoptosis rate and ROS level of HRECs were detected by using flow cytometry. MDA level and SOD activity were assessed by using enzyme-linked immunosorbent assay. miR-483-5p expression was detected by using Real-time quantitative PCR. Results Compared with the Con group, the apoptosis rate, ROS and MDA levels of HRECs in the HG group were significantly increased (P<0.05), SOD activity was significantly decreased (P<0.05), and miR-483-5p expression was significantly increased (P<0.05). Compared with the HG group, the apoptosis rate, ROS and MDA levels of HRECs in the HG+Mor-L group, HG+Mor-M group, and HG+Mor-H group were significantly reduced (P<0.05), and SOD activity was significantly increased (P<0.05), miR-483-5p expression was significantly reduced (P<0.05). Compared with the HG+anti-miR-NC group, the apoptosis rate, ROS and MDA levels of HRECs in the HG+anti-miR-483-5p group were significantly reduced (P<0.05), and SOD activity was significantly increased (P<0.05). Compared with the HG+Mor+miR-NC group, the apoptosis rate, ROS and MDA levels of HRECs in the HG+Mor+miR-483-5p group were significantly increased (P<0.05), and SOD activity was significantly decreased (P<0.05). Conclusions Morroniside inhibits high glucose-induced HRECs oxidative stress and apoptosis by down-regulating miR-483-5p expression.
关键词
莫诺苷;  /
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miR-483-5p;  /
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高糖;  /
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视网膜;  /
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血管内皮细胞;  /
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氧化应激;  /
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凋亡
Key words
Morroniside;  /
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miR-483-5p;  /
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High glucose;  /
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Retina;  /
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Vascular endothelial cells /
Oxidative stress;  /
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Apoptosis
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基金
国家自然科学基金委员会资助项目(31560292)