Abstract
Objective To investigate the effect of piperine on the inflammatory response of fibroblastoid synovial cells(HFLS-RA)in rheumatoid arthritis and its potential mechanism.Methods MH7A cells were randomly divided into control group(routine culture),tumor necrosis factor(TNF)-α group(cells were treated with 20 ng·mL-1 TNF-α),experimental-H group(100.00 μmol·L-1 piperine treatment),experimental-H+si-NC group(after transfection with the si-NC plasmid,piperine was combined with TNF-α for treatment),experimental-H+si-SFRP1(after transfection with the si-SFRP1 plasmid,piperine was combined with TNF-α for treatment)group.The relative expression levels of the relevant proteins were detected by Western blot and cell immunofluorescence methods respectively,flow cytometry was used to detect pyroptosis level,and the content of inflammatory factors was detected using a kit.Results The relative expression levels of SFRP1 protein in the control group,TNF-α group,experimental-H group,experimental-H+si-NC group and experimental-H+si-SFRP1 group were 1.06±0.11,0.35±0.03,0.93±0.04,0.91±0.11 and 0.26±0.03,respectively,when comparing cells in the TNF-α group with those in the control group,and comparing cells in the experimental-H+si-SFRP1 group with those in the experimental-H+si-NC group,the relative expression levels of SFRP1 protein were all significantly decreased(P＜0.05);compared with the TNF-α group,the protein level of SFRP1 was significantly increased in the experimental-H group(P＜0.05).The levels of pyroptosis the control group,TNF-α group,experimental-H group,experimental-H+si-NC group and experimental-H+si-SFRP1 group were(3.83±0.39)％,(25.24±1.34)％,(12.77±1.29)％,(12.36±1.03)％and(20.19±1.70)％,respectively;the levels of interleukin(IL)-6 were(10.01±1.09),(89.98±6.60),(20.31±2.72),(20.52±1.98)and(43.41±4.15)pg·mL-1,respectively;the relative expression levels of NOD-like receptor 3(NLRP3)protein were 0.25±0.06,0.75±0.08,0.43±0.07,0.39±0.05 and 0.64±0.10,respectively;the relative expression levels of β-catenin proteins were 0.28±0.03,0.92±0.12,0.46±0.07,0.44±0.06 and 0.58±0.10,the above indicators were significantly increased when compared between the TNF-α group and the control group,as well as between the experimental-H+si-SFRP1 group and the experimental-H+si-NC group(all P＜0.05);in contrast,these indicators were significantly decreased when compared between the experimental-H group and the TNF-α group(P＜0.05).Conclusion Piperine may regulate pyroptosis of MH7A cells mediated by the Wnt/β-catenin signaling pathway by upregulating SFRP1 expression,thereby inhibiting the inflammatory response.关键词
胡椒碱/类风湿关节炎/成纤维样滑膜细胞/细胞焦亡/分泌型卷曲相关蛋白1/炎性反应Key words
piperine/rheumatoid arthritis/fibroblast-like synovial cell/pyroptosis/secreted frizzled related protein 1/inflammatory response分类
医药卫生
