The Pathogenesis Of CD14 Engaging In Gout Of Spontaneous Remission

Research background With the growth in the living standard,the morbidity of gout has obviously increased.It has threated the health of human.An interesting feature of acute gout is the self-limiting nature of the inflammatory flare.In the absence of clinical intervention,a gouty episode will spontaneously resolve within 7-10 days.The mechanism of acute gout is complicated.There are PAMPs and DAMPs forms in human.TLRs and NLRs are two mainly molecules in PRRS.the deposition of monosodium urate(MSU)crystals in articular joints as foreign matters are the trigger to induce acute imflammatory reaction.But,the pathogenesis is not clearly.In gout,the deposition of monosodium urate(MSU)crystals in articular joints and bursal tissues can be asymptomatic or be associated with the pathogenesis of acute,episodic,selflimiting joint inflammationThe interaction of MSU crystals with synovial lining cells,macrophages,and other resident cells in the joint appears to be the primary trigger for the acute neutrophil ingress that drives episodes of gouty arthritis.The induction by MSU crystals of phagocyte release of mediators,including arachidonate metabolites,the cytokines TNF-α,IL-1 β,CXCL1(GROα),CXCL8(IL-8).MSU crystals are ingested by phagocytes.Significantly,MSU crystals functionally engage the canonical signaling pathway from TLR2 to NF-κB activation mediated by the shared TLR and IL-1 receptor adaptor protein MyD88.TLR2 and TLR4 each mediate macrophage uptake of the MSU crystal in vitro and MSU-crystal-induced induced inflammation in vivo.CD 14 is a GPI-anchored protein constitutively expressed on the surface of various cells,including monocytes,macrophages,polymorphonuclear neutrophils,Aside from this membrane-bound(mCD14)state,CD14 is also found in a circulating soluble(sCD14)state.The exact role of CD14 in physiological and pathological situations is not well defined.CD14 was initially described as a specific receptor for LPS,a compound from the outer cell wall of Gram-negative bacteria.At the monocyte surface,the LPS receptor mCD14 and the LPS-binding protein(LBP)interact with LPS and form a high affinity trimolecular complex that allows monocytes to detect the presence of LPS.The pattern recognition molecule CD 14 serves as a shared TLR2 and TLR4 adaptor molecule that increases TLR2-and TLR4-mediated functional responses to specific agonists,including peptidoglycan and LPS,respectively.CD 14 is essential for a full-blown MSU crystal-induced neutrophilic inflammatory response.So,exploring CD 14 molecules spontaneously in gout relief mechanism is expected to find a new targets for the treatment of gout.Research purpose Exploring CD14 molecules spontaneously in gout relief mechanism is expected to find a new targets for the treatment of gout.Methods sCD14was detected by ELISA in the serum of patients(n=40)with gout and healthy control(n=31)and we analyze the correlation between sCD 14 and CRP.We Collected peripheral blood from healthy control and then the PBMCs were isolated.The expression of CD 14 was detected by flow cytometry and the apoptotic cell of CD14+cell was detected after PBMC stimulated by MSU and LPS.Then,mRNA of CD 14 in gout patient was detected by Q-PCR.Results sCD14 decreased in gout patients(p=0.0001)Positive correlation between sCD14 and CRP in gout patients.Drecreased sCD14 in gout patients with CRP<9.PBMC from healthy volunteers was stimulated for 16h by LPS with 0μg/ml,0.1μg/ml,1μgg/ml,10μg/ml.CD14 expression was detected by flow cytomety.There is no difference to compare the data respectively analyzed by Mann-Whitney Utest.PBMC from healthy volunteers was stimulated for 16h by MSU with 0μg/ml,30μg/ml,100μg/ml,300μg/ml.CD14 expression was detected by flow cytomety.There is no difference between PBMC stimulated by 0μg/ml and 30μg/ml.However,obviously decreased CD 14 expression appeared in PBMC stimulated by MSU with 100μg/ml(p=0.0286)and 300μg/ml(p=0.0286).PBMC from healthy volunteers was stimulated for 16h by LPS with 0μg/ml、100ng/ml、1μg/ml、10μg/ml.sCD14 was detected by elisa.There is no difference to compare the data respectively analyzed by Mann-Whitney U test.PBMC from healthy volunteers was stimulated for 16h by MSU with 0μg/ml,30μg/ml,100μg/ml,300μg/ml.sCD14 was detected by elisa.There is no difference between PBMC stimulated by 0μg/ml and 30μg/ml.However,obviously decreased sCD14 appeared in PBMC stimulated byMSU with 100μg/ml(p=0.0296)and 300μg/ml(p=0.0278).PBMC from healthy volunteers was stimulated for 16h by LPS with 0μg/ml,0.1μg/ml,1μg/ml,10μg/ml.CD14 expression in CD68+ cell was detected by flow cytomety.There is no difference to compare the data respectively analyzed by Mann-Whitney U test.PBMC from healthy volunteers was stimulated for 16h by MSU with 0μg/ml,30μg/ml,100pg/ml,300μg/ml.CD14 expression in CD68+ cell was detected by flow cytomety.There is no difference between PBMC stimulated by 0μg/ml and 30μg/ml.However,obviously decreased CD 14 expression appeared in the CD68+cell of PBMC stimulated by MSU with 100μg/ml(p=0.0286)and 300μg/ml(p=0.0286).we did not find CD 14 in Intracellular cell and the number of apoptotic cells in CD 14+ cells was not increased.The reason of decreased sCD14 was not the combination of MSU and sCD14.However,the CD 14 expression of mRNA in gout patients increased(p=0.0252).Conclusion sCD14 decreased in gout patients and Positive correlation between sCD14 and CRP in gout patients.Drecreased sCD14 in gout patients with CRP<9.The CD 14 expression of PBMC has decreased after PBMC was stimulated by different concentration of MSU.Then,the CD14 expression of PBM was no difference after PBMC was stimulated by different concentration of LPS.At the same time,sCD14 decreased in supernatant of PBMC stimulated by different concentration of MSU.however,sCD14 was no difference after PBMC was stimulated by different concentration of LPS.we did not find CD 14 in Intracellular cell and the number of apoptotic cells in CD 14+ cells was not increased.The reason of decreased sCD 14 was not the combination of MSU and sCD14.However,the CD 14 expression of mRNA in gout patients increased.We speculat that there is other mechanisms of CD 14 decreased.