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AIM:To elucidate the anti-inflammatory mechanism ofan intestinal neuropeptide,sulfated cholecystokininoctapeptide(sCCK-8),the effects of sCCK-8 onIipopolysaccharide(LPS)-induced tumor necrosis factorα(TNF-α)mRNA expression and NF-κB activity inpulmonary interstitial macrophages(PIMs)werestudied.METHODS:PIMs from rat were stimulated with LPS(1mg·L~(-1))in the presence or absence of sCCK-8(10~(-8)-10~(-6)mol·L~(-1))or/and CCK receptor antagonistproglumide(2 mg·L~(-1)).The expression of TNF-a mRNAwas assayed by reverse transcription polymerase chainreaction(RT-PCR)at 3h of the stimulation,and nuclearfactor-κB(NF-κB)binding activity was analyzed byelectrophoretic mobility shift assay(EMSA)at 1 h ofstimulation.The IκB-α protein level in the cytoplasmaat 30 min of the stimulation was detected by Westernblot.RESULTS:sCCK-8,at concentrations from 10~(-8)mol·L~(-1)to 10~(-6)mol·L~(-1)obviously inhibited LPS-induced TNF-αmRNA expression and NF-κB binding activity in a dose-dependent manner,P<0.05,P<0.01.Stimulation PIMswith LPS resulted in a reduction of IκB-α protein level,P<0.01,which was elevated by sCCK-8,P<0.05.Theeffects of sCCK-8 on NF-κB activity and IκB protein levelwere attenuated by CCK receptor antagonistproglumide,P<0.01.CONCLUSION:sCCK-8 inhibits LPS-induced TNF-a mRNAexpression by regulating NF-κB activity in rat PIMs,which is mediated through CCK receptors and inhibiting IκB-α degradation.This represents one of the anti-inflammatory mechanisms of sCCK-8.
AIM: To elucidate the anti-inflammatory mechanism of an intestinal neuropeptide, sulfated cholecystokinin octapeptide (sCCK-8), the effects of sCCK-8 on Iipopolysaccharide (LPS) -induced tumor necrosis factor alpha (TNF- [alpha]) mRNA expression and NF-kB activity in pulmonary interstitial macrophages (PIMs) were stutied. METHODS: PIMs from rat were stimulated with LPS (1 mg · L -1) in the presence or absence of sCCK-8 The expression of TNF-a mRNA was assayed by reverse transcription polymerase chain reaction (RT-PCR) at 3h of the stimulation, and / or CCK receptor antagonistproglumide (2 mg · L -1) nuclear factor-κB (NF-κB) binding activity was analyzed by electrophophore mobility shift assay (EMSA) at 1 h ofstimulation.The IκB-α protein level in the cytoplasmaat 30 min of the stimulation was detected by Westernblot. RESULTS: sCCK-8, at concentrations from 10 ~ (-8) mol·L ~ (-1) to 10 ~ (-6) mol·L ~ (-1) obviously inhibited LPS-induced TNF-αmRNA expression and NF-κB binding activity in a dose- dependen t manner, P <0.05, P <0.01.Stimulation PIMs with LPS resulted in a reduction of IκB-α protein level, P <0.01, which was elevated by sCCK-8, P <0.05.The effects of sCCK-8 on NF- activity and IκB protein levelwere attenuated by CCK receptor antagonistproglumide, P <0.01.CONCLUSION: sCCK-8 inhibits LPS-induced TNF-a mRNA expression by regulating NF-κB activity in rat PIMs, which is mediated through CCK receptors and inhibiting IκB-α degradation .This represents one of the anti-inflammatory mechanisms of sCCK-8.