摘要
目的探讨积雪草苷对川崎病血管损伤的影响及其可能作用机制。方法采用肿瘤坏死因子-α(TNF-α)诱导人冠状动脉血管内皮细胞建立川崎病血管损伤模型,分为7组:con组、TNF-α组、TNF-α+低积雪草苷组、TNF-α+中积雪草苷组、TNF-α+高积雪草苷组、TNF-α+积雪草苷+pcDNA组、TNF-α+积雪草苷+pcDNA-MMP-9组;CCK-8法、流式细胞术分别检测细胞增殖及凋亡;qRT-PCR检测基质金属蛋白酶-9(MMP-9)mRNA的表达水平;ELISA法检测IL-6、IL-1的水平;Western blot法检测Bax、Bcl-2、MMP-9蛋白表达量。结果与con组比较,TNF-α组细胞凋亡率和Bax蛋白水平升高,IL-6、IL-1的水平升高,MMP-9 mRNA及蛋白水平升高,Bcl-2蛋白水平降低,差异均有统计学意义(P<0.05);与TNF-α组比较,TNF-α+低积雪草苷组、TNF-α+中积雪草苷组、TNF-α+高积雪草苷组细胞凋亡率和Bax蛋白水平降低,IL-6、IL-1的水平降低,MMP-9 mRNA及蛋白水平降低,Bcl-2蛋白水平升高,差异均有统计学意义(P<0.05),且呈剂量依赖性;与TNF-α+积雪草苷+pcDNA组比较,TNF-α+积雪草苷+pcDNA-MMP-9组MMP-9 mRNA及蛋白水平升高,细胞凋亡率和Bax蛋白水平升高,IL-6、IL-1的水平升高,Bcl-2蛋白水平降低,差异均有统计学意义(P<0.05)。结论积雪草苷可通过下调MMP-9表达而抑制TNF-α诱导的血管内皮细胞凋亡及炎症反应从而减轻川崎病血管损伤。
Objective To investigate the effect of aisaticoside on vascular injury in kawasaki disease and its possible mechanism. Methods Tumor necrosis factor-α(TNF-α) was used to induce human coronary vascular endothelial cells to establish a vascular injury model of kawasaki disease. The cells were divided into 7 groups: con group,TNF-α group, TNF-α+low aisaticoside group, TNF-α+medium aisaticoside group, TNF-α+high aisaticoside group,TNF-α+aisaticoside+pcDNA group, TNF-α+aisaticoside+pcDNA-MMP-9 group. Cell proliferation and apoptosis were detected by CCK-8 method and flow cytometry, respectively. The expression of matrix metalloproteinase-9(MMP-9)m RNA was detected by q RT-PCR. The levels of IL-6 and IL-1 were detected by ELISA. Western blot was used to detect the protein expressions of Bax, Bcl-2, and MMP-9. Results Compared with the con group, the apoptosis rate and the level of Bax protein in the TNF-α group were increased, the levels of IL-6 and IL-1 were increased, and the levels of MMP-9 mRNA and protein was increased, while the level of Bcl-2 protein was decreased, with statistically significant differences(P<0.05). Compared with the TNF-α group, the apoptosis rate and the protein level of Bax in the TNF-α+low aisaticoside group, the TNF-α + medium aisaticoside group, and the TNF-α + high aisaticoside group were decreased, the levels of IL-6 and IL-1 were decreased, the levels of MMP-9 mRNA and protein were decreased, and the protein level of Bcl-2 was increased, with statistically significant differences(P<0.05), in a dose-dependent manner.Compared with TNF-α+aisaticoside+pcDNA group, the mRNA and protein levels of MMP-9 in TNF-α+aisaticoside+pcDNA-MMP-9 group were increased, and the apoptosis rate and the protein level of Bax were increased, the levels of IL-6 and IL-1 were increased, while the protein level of Bcl-2 was decreased, with statistically significant differences(P<0.05). Conclusion Aisaticoside could reduce vascular injury in kawasaki disease by inhibiting TNF-α-induced apoptosis and inflammatory response of vascular endothelial cells by downregulating the expression of MMP-9.
作者
杨劲威
曹媛
YANG Jin-wei;CAO Yuan(Department of Pediatrics,Baoji People's Hospital,Baoji 721000,Shaanxi,CHINA;Department of Pharmacy,Baoji People's Hospital,Baoji 721000,Shaanxi,CHINA)
出处
《海南医学》
CAS
2023年第2期165-170,共6页
Hainan Medical Journal
关键词
积雪草苷
川崎病
基质金属蛋白酶-9
人冠状动脉血管内皮细胞
凋亡
炎症
Aisaticoside
Kawasaki disease
Matrix metalloproteinase-9
Human coronary vascular endothelial cells
Apoptosis
Inflammation
