RESUMO
Transient receptor potential melastatin 7 (TRPM7) is a cation channel that plays a role in the progression of rheumatoid arthritis (RA), yet its involvement in synovial hyperplasia and inflammation has not been determined. We previously reported that TRPM7 affects the destruction of articular cartilage in RA. Herein, we further confirmed the involvement of TRPM7 in fibroblast-like synoviocyte (FLS) proliferation, metastasis and inflammation. We observed increased TRPM7 expression in FLSs derived from human RA patients. Pharmacological inhibition of TRPM7 protected primary RA-FLSs from proliferation, metastasis and inflammation. Furthermore, we found that TRPM7 contributes to RA-FLS proliferation, metastasis and inflammation by increasing the intracellular Ca2+ concentration. Mechanistically, the PKCα-HuR axis was demonstrated to respond to Ca2+ influx, leading to TRPM7-mediated RA-FLS proliferation, metastasis and inflammation. Moreover, HuR was shown to bind to IL-6 mRNA after nuclear translocation, which could be weakened by TRPM7 channel inhibition. Additionally, adeno-associated virus 9-mediated TRPM7 silencing is highly effective at alleviating synovial hyperplasia and inflammation in adjuvant-induced arthritis rats. In conclusion, our findings unveil a novel regulatory mechanism involved in the pathogenesis of RA and suggest that targeting TRPM7 might be a potential strategy for the prevention and treatment of RA.
Assuntos
Artrite Experimental , Artrite Reumatoide , Proliferação de Células , Interleucina-6 , Proteína Quinase C-alfa , Sinoviócitos , Canais de Cátion TRPM , Canais de Cátion TRPM/metabolismo , Canais de Cátion TRPM/genética , Artrite Reumatoide/patologia , Artrite Reumatoide/metabolismo , Animais , Sinoviócitos/metabolismo , Sinoviócitos/patologia , Humanos , Interleucina-6/metabolismo , Interleucina-6/genética , Proteína Quinase C-alfa/metabolismo , Proteína Quinase C-alfa/genética , Artrite Experimental/patologia , Artrite Experimental/metabolismo , Masculino , Ratos , Fibroblastos/metabolismo , Fibroblastos/patologia , Proteína Semelhante a ELAV 1/metabolismo , Proteína Semelhante a ELAV 1/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Células Cultivadas , Inflamação/metabolismo , Inflamação/patologia , Ratos Sprague-Dawley , Feminino , Transdução de SinaisRESUMO
A direct and sensitive method for the detection of methyl centralite (MC) and ethyl centralite (EC) as gunshot residues (GSRs) has been developed. This method uses desorption electrospray ionization (DESI)-tandem mass spectrometry and directly desorbs and detects analytes from surfaces without any sampling process. Typical transitions for MC and EC, m/z 241 to m/z 134 and m/z 269 to m/z 148, respectively, were used to improve the assay sensitivity. It has been shown that MC and EC can be detected on various surfaces, with detection limits of 5-70 pg/cm(2). Interferences, detection time after shooting and the number of times hands were washed after shooting were also evaluated. None of the materials interfered with the results and the detection window for organic GSRs was up to 12 h and hands could be washed at least six times. Further samples were analyzed to confirm the reliability of this method, and showed that it could discriminate shooters from nonshooters. This method should be of significance in forensic science, especially in analyzing GSRs, because of its simplicity, high throughput, and the direct detection of MC and EC on suspects' hands, clothes, and hair.
