苦参碱对大鼠视网膜微血管内皮细胞的增生及VEGF表达的影响

Effect of matrine on cell proliferation and expression of VEGF in retinal microvascular endothelial cells of rat

目的探讨苦参碱(Ma)对体外培养的大鼠视网膜微血管内皮细胞的增生抑制及对血管内皮细胞生长因子(VEGF)表达的影响。方法原代培养大鼠视网膜微血管内皮细胞,噻唑蓝比色法(MTT)检测不同质量浓度的Ma对其增生的抑制作用。将培养的细胞分为对照组、Ma(40mg/L)组、高糖(25mmol/L)组、Ma+高糖组4组。免疫细胞化学和Westernblot法检测细胞VEGF的表达。结果一定质量浓度范围内Ma可抑制视网膜微血管内皮细胞的增生(P<0.05),48h内呈质量浓度依赖性。与对照组和高糖组相比,Ma组48h后,VEGF表达明显下降(P<0.05),与对照组相比,高糖组VEGF的表达明显增加(P<0.05)。结论Ma可抑制视网膜微血管内皮细胞的增生,抑制主要是通过下调VEGF表达而发生作用的,Ma对高糖引起的VEGF表达上调同样也有抑制作用。

Objective Neovascular retinopathy may lead to vision reduce or loss. Vascular endothelial growth factor is proved to be the key factor of retinal neovascularization. This study was to evaluate the effects of matrine on cell proliferation and expression of intercellular vascular epithelial growth factor（VEGF） in retinal microvascular endothelial cells in vitro. Methods The primary microvascular endothelial cells from rat retina were cultured and treated with 5,10,20,40,80 and 160 mg/L matrine for 48 hours. Cell viability was measured by MTT assay. The primary retinal microvascular endothelial cells were divided into control group,40 mg/L Ma group,25 mmol/L high glucose group and Ma（40 mg/L） ＋ high glucose （25 retool/L） group. After 48 hours, the localization of VEGF protein was examined by immunocytochemistry. Western blot was used to measure the protein level of VEGF. Results Cultured microvaseular endothelial cells showed polygonal and round in shape with the brown staining for factor Ⅷ. The absorbing optical value of cultured retinal microvascular endothelial cells was significant reduced in different concentrations of Ma treating groups at 48 hours in comparison with control group （ P 〈 0. 05 ）. The inhibiting rate of Ma to cultured retinal microvaseular endothelial cells was concentration-dependent （ P 〈 0.05 ）. Compared with the control group and high glucose group, Ma decreased the intercellular expression of VEGF. The expression of VEGF in intercellular tissue was 12.63 ± 4.16 in Ma treating group, indicating a obvious decrease compared with control group and high glucose group（26.42 ± 6.21, 38.24±1.82） （ P 〈 0.05 ）. However, in high glucose group, the expression of VEGF was significantly higher than the control group（P 〈 0.05）. Conclusion Ma suppresses the expression of VEGF in microvaseular endothelial cells in vitro,which may be one of important mechanisms to inhibit the proliferation of vascular endothelial cells.