蓬子菜总黄酮经调控TGF-β/Smad3信号通路对内皮细胞增殖、凋亡的影响研究

Study on the effect of total flavonoids of Brassica oleracea on the proliferation and apoptosis of endothelial cells by regulating TGF-β/Smad3 signal pathway

目的研究蓬子菜总黄酮经调控TGF-β/Smad3信号通路对人脐静脉内皮细胞(HUVECs)增殖、凋亡的影响。方法实验分为5组,对照组HUVECs正常培养,其余组HUVECs培养后建立H 2O 2氧化损伤模型,其中蓬子菜总黄酮低、中、高浓度组分别加入12.5 mg/L、25 mg/L、50 mg/L的蓬子菜总黄酮培养。采用MTT法检测细胞增殖情况,流式细胞仪检测细胞凋亡情况,酶联免疫法检测细胞中超氧化物歧化酶(SOD)、丙二醛(MDA)、内皮素-1(ET-1)、降钙素基因相关肽(CGRP)水平,Western blot法检测细胞中转化生长因子-β1(TGF-β1)、Smad3蛋白表达量。结果模型组及各浓度蓬子菜总黄酮组细胞增殖率及细胞中SOD、CGRP水平和TGF-β1蛋白表达量均明显低于对照组(P均<0.05),细胞凋亡率及细胞中MDA、ET-1水平和Smad3蛋白表达量均明显高于对照组(P均<0.05)。各浓度蓬子菜总黄酮组细胞中SOD、CGRP水平和TGF-β1蛋白表达量均明显高于模型组(P均<0.05),MDA、ET-1水平和Smad3蛋白表达量均明显低于模型组(P均<0.05)。中、高浓度蓬子菜总黄酮组细胞增殖率及细胞中SOD、CGRP水平和TGF-β1蛋白表达量均明显高于低浓度蓬子菜总黄酮组(P均<0.05),且高浓度蓬子菜总黄酮组均明显高于中浓度蓬子菜总黄酮组(P均<0.05);中、高浓度蓬子菜总黄酮组细胞凋亡率及细胞中MDA、ET-1水平和Smad3蛋白表达量均明显低于低浓度蓬子菜总黄酮组(P均<0.05),且高浓度蓬子菜总黄酮组均明显低于中浓度蓬子菜总黄酮组(P均<0.05)。结论蓬子菜总黄酮能促进HUVECs增殖,抑制其凋亡,可通过调控TGF-β/Smad3信号通路减轻氧化应激反应,改善血管功能,从而对HUVECs起到保护作用。

Objective It is to study the effect of total flavonoids of Brassica oleracea on the proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs)by regulating TGF-β/Smad3 signal pathway.Methods The experiment was divided into 5 groups.The HUVECs in the control group were cultured normally,and the HUVECs in the remaining groups were cultured to establish H 2O 2 oxidative damage models.Among them,the low,medium,and high concentration groups of total flavonoids of Brassica oleracea were respectively treated with 12.5 mg/L,25 mg/L,and 50 mg/L total flavonoids of Brassica oleracea.MTT method was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis,and enzyme-linked immunoassay was used to detect the levels of superoxide dismutase(SOD),malondialdehyde(MDA),endothelin-1(ET-1),calcitonin gene-related peptide(CGRP),Western blot was used to detect the expression of transforming growth factor-β1(TGF-β1)and Smad3 protein in the cells.Results The cell proliferation rate,the levels of SOD,CGRP and TGF-β1 protein expression in the cells of the model group and every total flavonoids of Brassica oleracea group were significantly lower than those in the control group(all P<0.05),and the apoptosis rate and the levels of MDA,ET-1 and Smad3 protein expression were significantly higher than those of the control group(all P<0.05).The levels of SOD,CGRP and TGF-β1 protein expression in the cells of each total flavonoids of Brassica oleracea group were significantly higher than those of the model group(all P<0.05),and the levels of MDA,ET-1 and Smad3 protein expression were significantly lower than those of the model group(all P<0.05).The cell proliferation rate and the levels of SOD,CGRP and TGF-β1 protein expression in the medium and high concentrations of total flavonoids of Brassica oleracea group were significantly higher than those in the low concentration group(P<0.05),and the high concentration group was significantly higher than the medium-concentration group(P<0.05);the apoptosis rate and the levels of MDA,ET-1 and Smad3 protein expression in the medium and high-concentration of total flavonoids of Brassica oleracea groups were significantly lower than those in the low-concentration group(P<0.05),and the high-concentration group were significantly lower than those in the medium-concentration group(all P<0.05).Conclusion Total flavonoids of Brassica oleracea can promote HUVECs cell proliferation and inhibit its apoptosis,improve oxidative stress response and vascular function by regulating TGF-β/Smad3 signal pathway,so as to protect HUVECs cells.