蓬子菜总黄酮基于TGF-β/Smad3和JNK信号通路对动脉硬化内皮细胞的影响研究  

Effects of flavone from Galium Verum L on arteriosclerosis endothelial cells via TGF-β/Smad3 signaling pathway

作　　者：徐丹[1] 刘思赢 彭湃 匡东旭 夏联恒 许贵军[1] 韩爽 XU Dan;LIU Siying;PENG Pai;KUANG Dongxu;XIA Lianheng;XU Guijun;HAN Shuang(The Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150000,Heilongjiang,China;The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150000 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《现代中西医结合杂志》2024年第11期1515-1519,1525,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：黑龙江省中医药管理局科研项目(ZHY18-069);黑龙江省中医药管理局中医药经典普及化研究项目(ZYW2022-031)。

摘　　要：目的探讨蓬子菜总黄酮基于转化生长因子-β(TGF-β)/Smad3和c-Jun氨基末端激酶(JNK)信号通路对动脉硬化内皮细胞的影响。方法将24只成年雄兔随机分为正常组、模型组、蓬子菜总黄酮低剂量组、蓬子菜总黄酮高剂量组,每组6只。除正常组外,其余组兔建立下肢动脉硬化闭塞模型。建模成功后,蓬子菜总黄酮低、高剂量组分别给予45 mg/kg、180 mg/kg的蓬子菜总黄酮灌胃,正常组和模型组给予等量生理盐水灌胃。各组兔灌胃2周后处死,取右侧髂股动脉,分离培养内皮细胞。采用Western blot法检测内皮细胞中TGF-β_(1)、Smad3、JNK及磷酸化JNK(p-JNK)蛋白表达情况,分别采用酶联免疫分析法、硝酸还原法、放射免疫法检测内皮细胞中炎症因子[C反应蛋白(CRP)、可溶性细胞间黏附因子-1(sICAM-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)]、一氧化氮(NO)、内皮素-1(ET-1)含量。结果模型组内皮细胞中TGF-β_(1)、Smad3、JNK、p-JNK蛋白表达量及CRP、sICAM-1、TNF-α、IL-1β、ET-1含量均明显高于正常组(P均<0.05),蓬子菜总黄酮各组上述各指标均明显低于模型组(P均<0.05),且蓬子菜总黄酮高剂量组各指标均明显低于蓬子菜总黄酮低剂量组(P均<0.05)。模型组内皮细胞中NO含量明显低于正常组(P<0.05),蓬子菜总黄酮各组NO含量均明显高于模型组(P均<0.05),且蓬子菜总黄酮高剂量组明显高于蓬子菜总黄酮低剂量组(P<0.05)。结论蓬子菜总黄酮可减轻炎症反应,保护血管内皮细胞,抑制平滑肌细胞增殖和细胞凋亡,改善血管内皮功能,机制可能与抑制TGF-β/Smad3和JNK信号通路有关。Objective It is to investigate the repair effect of flavone from Galium Verum L(FGVL)on arteriosclerosis endothelial cells based on transforming growth factor-β(TGF-β)/Smad3 and c-Jun amino-terminal kinase(JNK)signaling pathway.Methods Twenty-four adult male rabbits were randomly divided into normal group,model group,low-dose FGVL group and high-dose FGVL group,with 8 rabbits in each group.The rabbits of all groups except for the normal group were prepared to establish models of atherosclerotic occlusion of lower limbs.After successfully modeling,the low-dose FGVL group and high-dose FGVL group were given 45 mg/kg and 180 mg/kg of FGVL by gavage,respectively,the normal group and model group were given the same volume of normal saline by gavage.The rabbits in each group were gavaged for 2 weeks and then were executed,their right iliofemoral artery was taken to isolate and culture endothelial cells.The protein expressions of TGF-β_(1),Smad3,JNK and phosphorylated JNK(p-JNK)in the endothelial cells were detected by Western blot,and the contents of inflammatory factors[C-reactive protein(CRP),soluble intercellular adhesion molecule-1(sICAM-1),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)],nitric oxide(NO),and endothelin-1(ET-1)in the endothelial cells were respectively determined by enzyme-linked immunoassay,nitrate reduction,and radioimmunoassay.Results The protein expressions of TGF-β_(1),Smad3,JNK and p-JNK,and the contents of CRP,sICAM-1,TNF-α,IL-1βand ET-1 in the endothelial cells of the model group were significantly higher than those of the normal group(all P<0.05),the above indicators of the low-dose and high-dose FGVL groups were significantly lower than those of the model group(all P<0.05),and the above indicators of the high-dose FGVL group were significantly lower than those of the low-dose FGVL group(all P<0.05).The content of NO in the endothelial cells of the model group was significantly lower than that of the normal group(P<0.05),the content of NO in the endothelial cells of the low-

关 键 词：转化生长因子-β SMAD3 蓬子菜总黄酮 内皮细胞 

分 类 号：R-332[医药卫生]