| 都艳红,张士宏,夏春晓,等.基于磷脂酰肌醇 3激酶 /蛋白激酶 B通路探讨木犀草素对高糖诱导的视网膜血管内皮细胞损伤的影响[J].安徽医药,2026,30(4):676-680. |
| 基于磷脂酰肌醇 3激酶 /蛋白激酶 B通路探讨木犀草素对高糖诱导的视网膜血管内皮细胞损伤的影响 |
| Effects of luteolin on high glucose-induced retinal vascular endothelial cell injury based on the phosphatidylinositol 3-kinase/protein kinase B pathway |
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| DOI：10.3969/j.issn.1009-6469.2026.04.008 |
| 中文关键词: 木犀草素 糖尿病视网膜病变 高血糖 视网膜血管内皮细胞 血管生成 氧化应激 |
| 英文关键词: Luteolin Diabetic retinopathy Hyperglycemia Retinal vascular endothelial cell Angiogenesis Oxidative stress |
| 基金项目:保定市科技计划项目( 2141ZF218) |
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| 中文摘要: |
| 目的观察木犀草素对高糖诱导的视网膜血管内皮细胞( RVECs)损伤的影响，并探究可能的机制。方法该研究于 2023年 5月至 2024年 1月进行。 RVECs购自青旗(上海)生物技术有限公司，木犀草素购自西安弘康生物技术有限责任公司。体外常规培养 RVECs，用含有 30 mmol/L葡萄糖的 DMEM培养液处理建立损伤模型，分为高糖组(高糖处理)、木犀草素组(高糖处理 +30 μmol/L木犀草素)、 LY294002［磷脂酰肌醇 3激酶( PI3K)/蛋白激酶 B(AKT)通路阻断剂］组(高糖处理 +10 μmol/L LY294002)、联合组(高糖处理 +30 μmol/L木犀草素 +10 μmol/L LY294002)。另取不作任何处理的 RVECs为空白组。培养 24 h后，细胞计数试剂盒(CCK-8)法、膜联蛋白 V(Annexin V)/碘化丙啶( PI)双染法、血管拟态实验检测各组细胞活力、凋亡率、血管新生能力；检测细胞膜通透性及氧化应激指标；免疫印迹法检测各组 AKT、p-AKT、哺乳动物雷帕霉素靶蛋白( mTOR)p-mTOR蛋白表达量。结果与空白组比较，高糖组凋亡率［(2.87±0.45)%比( 10.15±3.45)%］、荧光强度( 0.21±0.06比 0.52±0.13)、、丙二醛含量［( 5.02±1.30)nmol/mg比( 18.87±3.65)nmol/mg］升高( P<0.05)，新生血管数量增加( P<0.05)，细胞存活率及 SOD、CAT活性降低( P<0.05)；与高糖组比较，木犀草素组凋亡率、荧光强度、丙二醛含量降低( P<0.05)细胞存活率升高( P<0.05)，新生血管数量减少( P<0.05)，SOD活性、 CAT活性增强( P<0.05)，LY294002组凋亡率、荧光强度、丙二，醛含量升高( P<0.05)，新生血管数量增加( P<0.05)SOD活性、 CAT活性降低( P<0.05)；与木犀草素组比较，联合组凋亡率、荧光强度、丙二醛含量升高( P<0.05)，新生血管数量增加( P<0.05)细胞存活率及 SOD活性、 CAT活性降低( P<0.05)；与 LY294002组比较，联合组凋亡率、荧光强度、丙二醛含量降低( P<0.05)细胞存活率升高( P<0.05)，新生血管数量减少( P<0.05)SOD活性、 CAT活性增强( P< 0.05)。与空白组比较，高糖组 p-AKT/AKT、p-mTOR/mTOR降低( P<0.05)；与高糖组比较，素组 p-AKT/AKT、p-mTOR/ 木犀草，mTOR升高( P<0.05)LY294002组 p-AKT/AKT、p-mTOR/mTOR降低( P<0.05)；与木犀草素组比较，联合组 p -AKT/AKT、 p -mTOR/mTOR降低(P<0.05)；与 LY294002组比较，联合组 p-AKT/AKT、p-mTOR/mTOR升高( P<0.05)。结论木犀草素可抑制高糖诱导的 RVECs凋亡及血管生成，增强细胞活力，降低细胞膜通透性，减轻氧化应激，从而保护细胞损伤，可能通过激活 PI3K/AKT通路来实现。 |
| 英文摘要: |
| Objective To observe the effect of luteolin on high glucose-induced retinal vascular endothelial cell (RVEC) injury and explore the possible mechanism.Methods This study was conducted from May 2023 to January 2024. RVECs were purchased fromQingqi (Shanghai) Biotechnology Co., Ltd., and luteolin was purchased from Xi'an Hongkang Biotechnology Co., Ltd. RVECs were cul-tured in vitro and treated with DMEM containing 30 mmol/L glucose to establish an injury model. The cells were divided into high glu-cose group (high glucose treatment), luteolin group (high glucose treatment + 30 μmmol/L luteolin), LY294002 [phosphatidylinositol 3-kinase (PI3K) / protein kinase B (AKT) pathway blocker] group (high glucose treatment + 10 μmol/L LY294002), and combinationgroup (high glucose treatment + 30 μmmol/L luteolin + 10 μmol/L LY294002). RVECs without any treatment were taken as blankgroup. After 24 h of culture, CCK-8 method, Annexin V/PI double staining method and vascular mimicry test were used to detect cell vi-ability, apoptosis rate and angiogenesis ability in each group, respectively. The cell membrane permeability and oxidative stress indica-tors were detected. Immunoblotting was used to detect the expression of AKT, p-AKT, mammalian target of rapamycin (mTOR), and p-mTOR proteins in each group.Results Compared with the blank group, the apoptosis rate [(2.87±0.45)% vs. (10.15±3.45)%], fluores-cence intensity (0.21±0.06 vs. 0.52±0.13), malondialdehyde (MDA) [(5.02±1.30) nmol/mg vs. (18.87±3.65) nmol/mg] content increased (P<0.05), the number of newly formed blood vessels increased (P<0.05), cell survival rate, the activities of superoxide dismutase (SOD) and catalase (CAT) decreased in the high glucose group (P<0.05). Compared with the high glucose group, the apoptosis rate, fluores-cence intensity, MDA content decreased (P<0.05), cell survival rate increased (P<0.05), the number of newly formed blood vessels de-creased (P<0.05), the activities of SOD and CAT increased in the luteolin group (P<0.05), the apoptosis rate, fluorescence intensity, MDA content increased (P<0.05), the number of newly formed blood vessels increased (P<0.05), the activities of SOD and CAT de-creased in the LY294002 group (P<0.05). Compared with the luteolin group, the apoptosis rate, fluorescence intensity, MDA content in-creased (P<0.05), the number of newly formed blood vessels increased (P<0.05), cell survival rate, the activities of SOD and CAT de-creased in the combination group (P<0.05). Compared with the LY294002 group, the apoptosis rate, fluorescence intensity, MDA con-tent decreased (P<0.05), cell survival rate increased (P<0.05), the number of newly formed blood vessels decreased (P<0.05), the activi-ties of SOD and CAT increased in the combination group (P<0.05). Compared with the blank group, p-AKT/AKT and p-mTOR/mTOR decreased in the high glucose group (P<0.05). Compared with the high glucose group, p-AKT/AKT and p-mTOR/mTOR increased in the luteolin group (P<0.05), p-AKT/AKT and p-mTOR/mTOR decreased in the LY294002 group (P<0.05). Compared with the luteolin group, p-AKT/AKT and p-mTOR/mTOR decreased in the combination group (P<0.05). Compared with the LY294002 group, p-AKT/ AKT and p-mTOR/mTOR increased in the combination group (P<0.05).Conclusion Luteolin can inhibit the apoptosis and angiogene-sis of RVECs induced by high glucose, enhance cell viability, reduce the cell membrane permeability, alleviate the oxidative stress, andprotect the cell injury, which may be achieved by activating the PI3K/AKT pathway. |
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