羟基红花黄色素A通过抑制LPS刺激的NLRP3炎性小体的激活抑制ANG Ⅱ诱导的VAFs的迁移

Hydroxysafflor yellow A inhibits the migration of vascular adventitial fibroblasts induced by ANG I via inhibiting LPS-stimulated NLRP3 inflammasome activation

本文探讨了羟基红花黄色素A(hydroxysafflor yellow A,HSYA)通过抑制NLRP3炎性小体的激活抑制血管紧张素Ⅱ(angiotensinⅡ,ANGⅡ)诱导的血管外膜成纤维细胞(vascular adventitial fibroblasts,VAFs)迁移的影响。采用组织贴块法培养原代VAFs,激光共聚焦技术鉴定VAFs;划痕实验测定HSYA对ANGⅡ诱导的VAFs迁移能力的影响;CCK-8实验测定HSYA对细胞活力的影响;Western blot实验检测各组细胞中NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)炎性小体相关蛋白的表达。激光共聚焦结果显示,VAFs的Vimentin表达为阳性,证实组织贴块培养的细胞为VAFs;划痕实验结果显示,脂多糖(lipolyaccharide,LPS)显著增强了ANGⅡ诱导的VAFs迁移;HSYA显著抑制ANGⅡ及LPS诱导的VAFs的迁移。Western blot结果显示,HSYA显著抑制ANGⅡ及LPS激活的NLRP3炎性小体相关蛋白NLRP3、含半胱氨酸的天冬氨酸蛋白水解酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)及凋亡相关斑点样蛋白(apoptosis-associated specklike protein containing CARD,ASC)蛋白表达。综上所述,HSYA通过抑制LPS刺激的NLRP3炎性小体激活,降低了ANGⅡ诱导的VAFs迁移。

This work is aimed to investigate whether hydroxysafflor yellow A(HSYA)inhibits the migration of vascular adventitial fibroblasts(VAFs)induced by angiotensin I(ANG II)via inhibiting the activation of NLRP3 inflammasome.VAFs were cultured by tissue paste method and the laser confocal technique was used to determine VAFs.The effect of HSYA on ANG II induced VAFs migration was determined by scratch test.CCK-8 assay was used to measure the effect of HSYA on cell viability.Western blot assay was used to detect the expression of NLRP3 inflammasome-related proteins.Laser confocal results showed that the cells cultured by tissue patch were VAFs.Scratch test results indicated that the migration rate of cells in ANG II group was significantly increased.HSYA significantly inhibited the migration of VAFs induced by ANG II or LPS.Western blot results showed that the expression of NLRP3 inflammasome related proteins NLRP3,Caspase-1 and ASC were significantly inhibited in HSYA group induced by ANG II or LPS.In conclusion,HSYA inhibits the migration of VAFs induced by ANG II via inhibiting the activation of NLRP3 inflammasome stimulated by LPS.