白杨素通过下调 VEGF 和 VEGFR 抑制高糖诱导的脉络膜视网膜内皮细胞迁移。
Chrysin Inhibits High Glucose-Induced Migration on Chorioretinal Endothelial Cells via VEGF and VEGFR Down-Regulation.
机构信息
Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei 111, Taiwan.
Department of Ophthalmology, Tri-Service General Hospital, National Defense Medical Center, Taipei 11490, Taiwan.
出版信息
Int J Mol Sci. 2020 Aug 2;21(15):5541. doi: 10.3390/ijms21155541.
BACKGROUND
Diabetes mellitus (DM) is a chronic inflammatory disease, which causes multiple complications. Diabetic retinopathy (DR) is among these complications and is a dominant cause of vision loss for diabetic patients. Numerous studies have shown that chrysin, a flavonoid, has many biological activities such as anti-oxidation and anti-inflammation. However, it is rarely used in ocular diseases. In this study, we examined the inhibitory effects of flavonoid on high glucose induced migration of chorioretinal endothelial cells (RF/6A cells) and its mechanism.
MATERIALS AND METHODS
The viability of RF/6A cells treated with chrysin was examined with a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The migration of RF/6A cells was assessed by the transwell migration and scratch wound assays. The expression of AKT, ERK, vascular endothelial growth factor (VEGF), HIF-1α and MMP-2 were determined by western blotting. To observe the mRNA expression of VEGF receptor (VEGFR), qRT-PCR, was utilized.
RESULTS
The results showed that chrysin can dose-dependently inhibit the RF/6A cell migration in vitro transwell and the scratch wound assays which are induced by high glucose. After pretreatment of RF/6A cells with different concentrations of chrysin, they did not produce any cytotoxicity in MTT assay. Moreover, chrysin down-regulated both phosphorylated AKT and ERK, as well as attenuated the expression levels of MMP-2. It also decreased the expression of the VEGF transcription factor and VEGF. Furthermore, it was shown that chrysin could suppress the protein and mRNA expression levels of VEGFR.
CONCLUSION
The results indicate that chrysin could down-regulate the phosphorylation of AKT, ERK and MMP-2 and reduce the effects of VEGF and VEGFR in a high glucose environment. It further inhibits the high glucose-induced migration of RE/6A cells. Therefore, chrysin may have the potential for visual protection.
背景
糖尿病（DM）是一种慢性炎症性疾病，会导致多种并发症。糖尿病视网膜病变（DR）是其中的并发症之一，也是糖尿病患者视力丧失的主要原因。大量研究表明，黄酮类化合物白杨素具有抗氧化和抗炎等多种生物活性。然而，它在眼部疾病中很少被使用。在这项研究中，我们研究了黄酮类化合物对高糖诱导的脉络膜视网膜内皮细胞（RF/6A 细胞）迁移的抑制作用及其机制。
材料与方法
通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐（MTT）测定法检测白杨素处理的 RF/6A 细胞的活力。通过 Transwell 迁移和划痕伤口测定法评估 RF/6A 细胞的迁移。通过 Western blot 测定 AKT、ERK、血管内皮生长因子（VEGF）、HIF-1α 和 MMP-2 的表达。利用 qRT-PCR 观察 VEGF 受体（VEGFR）的 mRNA 表达。
结果
结果表明，白杨素可剂量依赖性地抑制体外高糖诱导的 RF/6A 细胞迁移 Transwell 和划痕伤口试验。在 RF/6A 细胞用不同浓度的白杨素预处理后，它们在 MTT 测定中没有产生任何细胞毒性。此外，白杨素下调了磷酸化 AKT 和 ERK 的表达，并减弱了 MMP-2 的表达水平。它还降低了 VEGF 转录因子和 VEGF 的表达。此外，研究表明，白杨素可以抑制 VEGFR 的蛋白和 mRNA 表达水平。
结论
结果表明，白杨素可下调 AKT、ERK 和 MMP-2 的磷酸化，并减少高糖环境中 VEGF 和 VEGFR 的作用，从而抑制 RE/6A 细胞的高糖诱导迁移。因此，白杨素可能具有视觉保护作用。
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