Activation of mitogen-activated protein kinases in satellite glial cells of the trigeminal ganglion contributes to substance P-mediated inflammatory pain

Inflammatory orofacial pain,in which substance P (SP) plays an important role,is closely related to the cross-talk between trigeminal ganglion (TG) neurons and satellite glial cells (SGCs).SGC activation is emerging as the key mechanism underlying inflammatory pain through different signalling mechanisms,including glial fibrillary acidic protein (GFAP) activation,phosphorylation of mitogen-activated protein kinase (MAPK) signalling pathways,and cytokine upregulation.However,in the TG,the mechanism underlying SP-mediated orofacial pain generated by SGCs is largely unknown.In this study,we investigated whether SP is involved in inflammatory orofacial pain by upregulating interleukin (IL)-1β and tumour necrosis factor (TNF)-α from SGCs,and we explored whether MAPK signalling pathways mediate the pain process.In the present study,complete Freund’s adjuvant (CFA) was injected into the whisker pad of rats to induce an inflammatory model in vivo.SP was administered to SGC cultures in vitro to confirm the effect of SP.Facial expression analysis showed that pre-injection of L703,606 (an NK-1 receptor antagonist),U0126 (an inhibitor of MAPK/extracellular signal-regulated kinase [ERK] kinase [MEK] 1/2),and SB203580 (an inhibitor of P38) into the TG to induce targeted prevention of the activation of the NK-1 receptor and the phosphorylation of MAPKs significantly suppressed CFA-induced inflammatory allodynia.In addition,SP promoted SGC activation,which was proven by increased GFAP,p-MAPKs,IL-1β and TNF-α in SGCs under inflammatory conditions.Moreover,the increase in IL-1β and TNF-α was suppressed by L703,606,U0126 and SB203580 in vivo and in vitro.These present findings suggested that SP,released from TG neurons,activated SGCs through the ERK1/2 and P38 pathways and promoted the production of IL-1β and TNF-α from SGCs,contributing to inflammatory orofacial pain associated with peripheral sensitization.

Determination of Enantiomers in ET-26-HCl by Ultra Performance Convergence Chromatography

[Objectives]To establish an ultra performance convergence chromatography(UPC2)method to determine the content of enantiomers in(R)-2-methoxyethyl1-(1-phenylethyl)-1H-imidazole-5-carboxylate hydrochloride(ET-26 HCl).[Methods]ChiralpakAD-3 chromatographic column(4.6 mm×100 mm,3μm)was used,with 0.1%diethylamine methanol solution-supercritical carbon dioxide(1∶9)as mobile phase;flow rate 2.0 mL/min;back pressure 2000 psi,detection wavelength 240 nm,and column temperature 35℃.[Results]ET-26-HCl was completely separated from the enantiomers,and the linear relationship was good;the detection limit was 1.5μg/mL.[Conclusions]This method is suitable for the determination of the content of enantiomers in ET-26-HCl.