金铁锁三萜类成分对衣霉素诱导RA-FLS细胞内质网应激的影响

Effects of triterpenoids from Psammosilene tunicoides on tunicamycin-induced endoplasmic reticulum stress in RA-FLS

目的基于内质网通路探索金铁锁三萜类成分皂皮酸、丝石竹皂苷元对衣霉素诱导类风湿性关节炎成纤维细胞(RA-FLS)的作用机制。方法以衣霉素诱导RA-FLS细胞为研究对象,给予皂皮酸、丝石竹皂苷元进行干预,检测细胞增殖活性,ELISA法检测肿瘤坏死因子-α(TNF-α)水平,流式细胞术检测细胞凋亡,Transwell实验检测细胞迁移能力,Western blot法检测细胞中转录激活因子6(ATF-6)、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、半胱氨酸蛋白酶蛋白-12(caspase-12)、抗凋亡Bcl-2蛋白表达,RT-qPCR法检测细胞中ATF-6、GRP78、CHOP mRNA表达。结果与模型组比较,皂皮酸、丝石竹皂苷元各剂量组RA-FLS细胞TNF-α水平均降低(P<0.01),细胞增殖、迁移能力均减弱(P<0.01),凋亡率均升高(P<0.01),细胞ATF-6、Bcl-2蛋白表达降低(P<0.05,P<0.01),CHOP、caspase-12蛋白表达升高(P<0.05,P<0.01);皂皮酸、丝石竹皂苷元低、中剂量组GRP78蛋白表达降低(P<0.05,P<0.01);皂皮酸、丝石竹皂苷元中剂量组ATF-6、GRP 78 mRNA表达降低(P<0.01),CHOP mRNA表达升高(P<0.01)。结论皂皮酸、丝石竹皂苷元可能通过调节内质网信号通路,降低相关炎症因子分泌,抑制RA-FLS细胞增殖及迁移,诱导细胞凋亡,起到对类风湿性关节炎的保护作用。

AIM To investigate the mechanism of triterpenoids quillaic acid and gypsogenin-3-O-glucuronide from Psammosilene tunicoides on tunicamycin-induced rheumatoid arthritis fibroblasts-like synoviocytes(RA-FLS)via the endoplasmic reticulum pathway.METHODS The research objects of tunicamycin-induced RA-FLS intervened with quillaic acid and gypsogenin-3-O-glucuronide had their cell proliferation activity detected;their level of tumor nerosis factor-α(TNF-α)detected by ELISA;their apoptosis detected by flow cytometry;their cell migration ability detected by Transwell experiment;their expressions of transcription activator 6(ATF-6),glucose regulatory protein 78(GRP78),C/EBP homologous protein(CHOP),cysteine protease protein-12(caspase-12)and anti-apoptosis Bcl-2 protein detected by Western blot;and their mRNA expressions of ATF-6,GRP 78 and CHOP detected by RT-qPCR.RESULTS Compared with the model group,each group intervened with quillaic acid or gypsogenin-3-O-glucuronide displayed decreased levels of TNF-α(P<0.01);weakened cell proliferation and migration ability(P<0.01);increased apoptosis rate(P<0.01);decreased protein expressions of ATF-6 and Bcl-2(P<0.05,P<0.01);and increased protein expressions of CHOP and caspase-12(P<0.05,P<0.01).In addition,decreased GRP78 protein expression in the low and medium dose groups(P<0.05,P<0.01);decreased mRNA expression of ATF-6,GRP78(P<0.01)and increased CHOP mRNA expression(P<0.01)in the medium dose groups of quillaic acid and gypsogenin-3-O-glucuronide were observed as well.CONCLUSION Quillaic acid and gypsogenin-3-O-glucuronide may play a protective role in rheumatoid arthritis by inhibiting the proliferation and migration of RA-FLS,inducing apoptosis and reducing the secretion of related inflammatory factors via endoplasmic reticulum signal pathway.