牛蒡子苷调控血管内皮生长因子抑制糖尿病视网膜病变的机制研究

Arctiin inhibits diabetic retinopathy through regulating vascular endothelial growth factor(VEGF)

目的探讨牛蒡子苷(Arc)是否通过调控血管内皮生长因子(VEGF)表达进而抑制糖尿病视网膜病变。方法体外培养人视网膜微血管内皮细胞(HRCECs),用不同浓度的Arc处理HRCECs,同时将si-VEGF或pcDNA3.1-VEGF分别转染入HRCECs,MTT法检测HRCECs增殖能力;Western blot检测p21、细胞周期蛋白D1(cyclinD1)、肿瘤坏死因子-α(TNF-α)、人单核细胞趋化蛋白-1(MCP-1)、载脂蛋白M(ApoM)、VEGF蛋白表达。结果与LG组相比,HG组HRCECs的OD值明显升高(P<0.05),cyclinD1、TNF-α、MCP-1、ApoM、VEGF水平均显著升高(P<0.05),而p21表达下调(P<0.05),Arc作用后可显著逆转HG对HRCECs增殖、cyclinD1、TNF-α、MCP-1、ApoM、VEGF及p21表达的作用,且Arc不同剂量组间均存在显著性差异(P<0.05);抑制VEGF表达可明显减弱高糖作用下的HRCECs增殖能力并降低相关炎性因子表达;过表达VEGF能逆转Arc对高糖诱导的HRCECs增殖及炎症反应的抑制作用。结论牛蒡子苷可能通过降低高糖诱导的HRCECs细胞中VEGF的高表达进而抑制HRCECs增殖,并可通过减轻炎症反应保护细胞免受损伤。

Objective We aim to investigate whether arctiin(Arc)inhibits diabetic retinopathy by regulating the expression of vascular endothelial growth factor(VEGF).Methods Human retinal microvascular endothelial cells(HRCECs)were cultured in vitro,and HRCECs were treated with different concentrations of Arc,and si-VEGF or pcDNA3.1-VEGF were transfected into HRCECs.The proliferation of HRCECs was detected by MTT assay.The expressions of p21,cyclinD1,TNF-α,MCP-1,ApoM and VEGF protein were detected by Western blot.Results Compared with LG group,the OD value of HRCECs in HG group was significantly increased(P<0.05),and cyclinD1,TNF-α,MCP-1,ApoM,VEGF levels were significantly increased(P<0.05),while p21 expression was down-regulated(P<0.05).After Arc treatment,the effect of HG on the proliferation of HRCECs,cyclinD1,TNF-α,MCP-1,ApoM,VEGF and p21 was significantly reversed,and there were significant differences between different doses of Arc(P<0.05).Inhibition of VEGF expression significantly attenuated the proliferation of HRCECs under high glucose and decreased the expression of related inflammatory factors.Overexpression of VEGF reversed the inhibitory effect of Arc on high glucose-induced proliferation and the inflammatory response of HRCECs.Conclusion Arc inhibits the proliferation of HRCECs by reducing the high expression of VEGF in HRCECs induced by high glucose,and may protect cells from damage by reducing inflammation.