丹皮酚通过调节中脑星形细胞神经营养因子的表达及细胞质/核易位抑制类风湿关节炎成纤维滑膜细胞炎症反应

Paeonol suppresses fibroblast synovitis by increasing MANF expression

目的探讨丹皮酚抑制类风湿关节炎成纤维样滑膜细胞(RA-FLSs)的炎症反应可能的机制。方法人成纤维样滑膜细胞(FLSs)通过肿瘤坏死因子(TNF-α)刺激及丹皮酚处理后,利用CCK-8法检测丹皮酚对RA-FLSs炎性增殖的抑制水平;Western blot检测MANF与内质网应激相关蛋白ATF6变化水平;Western blot及免疫荧光实验检测丹皮酚对MANF细胞质/核易位的影响;实时荧光定量分析PCR(RT-qPCR)法检测丹皮酚对转录因子p65的转录活性的影响。结果丹皮酚抑制RA-FLSs的异常增殖及迁移;丹皮酚促进内质网应激相关蛋白ATF6及MANF的表达增加;丹皮酚促进MANF蛋白向细胞核转移;丹皮酚能够剂量依赖性地抑制p65转录活性。结论丹皮酚通过内质网应激途径,促进MANF的表达及核转移,通过抑制p65转录活性,抑制RA-FLSs的炎症反应。

Aim To investigate the possible mechanism of paeonol inhibiting the inflammatory response of fibroblast synovial cells(RA-FLSS)in rheumatoid arthritis.Methods CCK-8 assay was used to detect Paeonol′s inhibitory level on the abnormal proliferation of arthritis human fibroblast synovial cells(RA-FLSs).The levels of endoplasmic reticulum stress-related proteins MANF and ATF6 were detected by Western blot.Cell localization of transcription factor p65 and Mesencephalic Astrocyte Derived Neurotrophic Factor(MANF)was detected by immunofluorescence.RT-qPCR detected the changes of p65 target genes.Results Paeonol could significantly inhibit the abnormal proliferation of RA-FLSS cells.Paeonol activates ATF6 and increases the expression of MANF.Paeonol promoted the nuclear transfer of MANF protein and inhibited the transcriptional activity of p65.Conclusion Paeonol promotes the expression of MANF and nuclear transfer through the endoplasmic reticulum stress pathway and affects the progression of RA by inhibiting the transcriptional activity of p65.