舒肝明目汤含药血清调节HIF1α-VEGF-Notch1通路活性影响脉络膜血管新生作用的研究

Effect of medicated serum prepared with Shugan Mingmu decoction on choroidal neovascularization by HIF1α-VEGF-Notch1 pathway

关键词:  舒肝明目汤  年龄相关的黄斑变性  脉络膜新生血管  脉络膜内皮细胞  低氧诱导因子1α

Key words:  medicated serum prepared with Shugan Mingmu decoction  age-related macular degeneration  choroidal neovascularization  choroidal endothelial cell  hypoxia-inducible factor 1

摘要:

[目的] 探讨舒肝明目汤含药血清对大鼠脉络膜血管内皮细胞（CECs）增殖的影响及与HIF1α-VEGF-Notch1通路相关的调节机制。[方法] CECs细胞在常氧和低氧条件下进行培养，检测低氧条件下舒肝明目汤含药血清对CECs增殖移行能力的影响；并采用实时荧光定量PCR法（RT-PCR）和免疫印迹法（Western blot）技术检测细胞中低氧诱导因子1α（HIF1α）、血管内皮生长因子（VEGF）和Notch1 mRNA和蛋白表达的水平。[结果] 与正常组相比，低氧组CECs的增殖移行能力明显提高（P<0.01），CECs中HIF1α、VEGF和Notch1 mRNA及蛋白表达明显增多（P<0.01）.经含药血清处理后，CECs增殖移行能力较低氧组显着下降（P<0.01），同时HIF1α和VEGF mRNA及蛋白表达降低（P<0.01，P<0.05），而Notch1表达进一步增高（P<0.01）.[结论] 舒肝明目汤含药血清通过上调Notch1表达及下调HIF1α-VEGF通路活性抑制CECs增殖迁移，从而减少脉络膜新生血管生成。

Abstract:

[Objective] In this study, the potential effect of medicated serum prepared with Shugan Mingmu decoction on proliferation ability of rat choroidal endothelial cells (CECs) and HIF1α-VEGF-Notch1 dependent regulatory mechanisms were investigated. [Methods] CECs cells were cultured in normoxic and hypoxic conditions. The effects of medicated serum prepared with Shugan Mingmu decoction on proliferation and migration abilities of CECs were detected. The expression levels of HIF1α, VEGF and Notch1 mRNA and protein were determined by RT-PCR and western blot. [Results] Compared with the control group, the proliferation and migration abilities of CECs in the hypoxia group were significantly promoted (P<0.01), and significant increased HIF1α, VEGF and Notch1 mRNA and protein expression were also detected (P<0.01). After treated with medicated serum, the migration and migration abilities of CECs decreased obviously when compared with the hypoxia group (P<0.01). The mRNA and protein expression levels of HIF1α and VEGF were decreased (P<0.01 and P<0.05), whereas the expression of Notch1 was increased further (P<0.01). [Conclusion] Medicated serum prepared with Shugan Mingmu decoction could inhibit CECs proliferation and migration by up-regulating the expression of Notch1 and down-regulating the activity of HIF1α-VEGF signal pathway, and thus decreased the formation of choroidal neovascularization.