骆驼蓬抑制高糖诱导的内皮细胞管道形成的作用机制研究

The mechanism by which Peganum harmala L. inhibits high glucose-induced tube formation in endothelial cells

探讨骆驼蓬对高糖诱导的视网膜血管内皮细胞管道形成的调节作用及其机制。建立高糖诱导的视网膜血管内皮细胞模型,将细胞分为:正常组(葡萄糖浓度为5.5 mmol·L^(-1))、模型组(葡萄糖浓度为25 mmol·L^(-1))、骆驼蓬给药高、低剂量组(葡萄糖浓度为25 mmol·L^(-1)+不同剂量骆驼蓬)。内皮管道形成方法观察细胞管道形成状态。网络药理学方法筛选骆驼蓬治疗糖尿病视网膜病变的药效成分、靶点和通路。Real-time PCR法验证骆驼蓬治疗糖尿病视网膜病变相关靶点的mRNA表达水平。内皮管道形成实验结果显示,与正常组相比,模型组内皮细胞管道形成总长度显著增加;与模型组相比,骆驼蓬组内皮管道形成总长度显著减少。网络药理学结果提示,骆驼蓬的作用靶点是细胞外调节蛋白激酶2 (extracellular signal-regulated kinase 2, ERK2)、丝氨酸苏氨酸蛋白激酶1 (serine/threonineprotein kinase 1, AKT1)、磷脂酰肌醇3-激酶催化亚基α(phosphoinositide 3 kinase catalytic alpha polypeptide,PIK3CA)等;信号通路涉及血管内皮生长因子(vascular endothelial growth factor, VEGF)信号通路和丝裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)信号通路等。Real-time PCR结果显示,与正常组相比,模型组ERK2、PIK3CA和AKT1的mRNA表达水平均升高;与模型组相比,骆驼蓬组ERK2、PIK3CA和AKT1的mRNA表达水平均降低。骆驼蓬可能通过调节MAPK信号通路和VEGF信号通路等抑制高糖条件下视网膜血管内皮细胞管道的形成,证实了它多靶点、多通路的作用特点。此研究为骆驼蓬治疗糖尿病视网膜病奠定了工作基础。

The study investigates the mechanism by which Peganum harmala L.(Luotuopeng, LTP) inhibits tube formation in retinal vascular endothelial cells. Tube formation was induced by treatment of retinal vascular endothelial cells with glucose. The cells were divided into a normal group, model group, and an LTP group. The total length of tube formation was measured. The active components, targets, and pathway by which LTP acts in the treatment of diabetic retinopathy was explored by network pharmacology. The mRNA expression levels of targets[extracellular signal-regulated kinase 2(ERK2), phosphoinositide 3 kinase catalytic alpha polypeptide(PIK3CA),serine/threonine-protein kinase 1(AKT1)] related to the mitogen-activated protein kinase(MAPK) signaling pathway and vascular endothelial growth factor(VEGF) signaling pathway was measured by real-time PCR. The results of tube formation indicated that compared with the normal group, the total tube length increased in the model group(P < 0.01);after the treatment with LTP, the total tube length decreased compared with the model group(P < 0.01). Network pharmacology revealed that the targets of LTP included PIK3CA, AKT1, and ERK2, and the pathways involved the MAPK signaling pathway and the VEGF signaling pathway. Real-time PCR indicated that compared with the normal group, the mRNA expression levels of ERK2, PIK3CA and AKT1 were elevated in the model group(P < 0.05);after treatment with LTP, the mRNA expression levels of ERK2, PIK3CA and AKT1 decreased compared with the model group(P < 0.05). LTP may inhibit retinal vascular endothelial cell tube formation by regulating the MAPK signaling pathway and the VEGF signaling pathway. This study confirms the multi-targets and multi-pathways of LTP and provides a basis for its use in the treatment of diabetic retinopathy.