The Fast Track To Canonical Wnt Signaling In MC3T3-E1 Cells Protected By Substance P Against Apoptosis

In clinical practice,osteosynthesis is the critical risk factor that always delaye fracture healing leading to bone nonunion and other complications,which seriously affect the quality of life of patients.Nowadays,in the research field focusing on fracture healing,techniques such as tissue engineered bone and external fixation have been developing rapidly,which has greatly improved the prognosis of patients.Reasoning its development,have benefited from the complex fracture regeneration and reconstruction of the relevant mechanisms of research.Thus,the biological mechanis,m of bone healing in the study,for the clinical treatment of innovation is important.During fracture healing,the role of neurological factors are highlighted.Recent studies have shown that neuropeptide neurotransmitter factors,such as calcitonin gene-related peptide(CGRP)and substance P(Substanc P,SP),can be secreted by peripheral nerves.In this study,we examined the effect of substance P on the apoptosis of MC3T3-E1 osteoblasts induced by serum deprivation,and explored the mechanisms involved in the expression of NK-1 receptor,the relationship between serum deprivation and classic Wnt pathway,as well as the role of Wnt Pathway in inducing apoptosis of Substance P in MC3T3-E1 Osteoblasts.Purpose:(1)mouse MC3T3-E1 osteoblast culture and NK-1 receptor expression during serum f'ree induced apoptosis;(2)apoptosis for mice MC3T3-E1 osteoblasts;(3)the biological effects of substance P on serum free induced apoptosis in mouse MC3T3-E1 osteoblasts;(4)the effect of substance P on MC3T3-E1 regulating NK-1 Receptor and Classical Wnt Pathway in Biological Effect of Induced Serum-induced apoptosis in Bone Cells.The aim of this study was to investigate the biological effects of these three factors on MC3T3-E1 cells by studying the biological effects of substance P(including its receptor NK-1),classical Wnt signaling pathway,and serum-induced apoptosis.Changes in NK-1 receptor levels during apoptosis were also examined.Method:to use experimental cell culture,MC3T3-E1 osteoblast chemical staining identification,PCR detection of target gene expression,Western Blot detection of target protein expression,immunofluorescent labeling-related antibody-FITC staining and other methods,The degree of cell gene transduction and the expression of protein and so on,elucidate the mechanism of action of neuropeptide substance P.Result:osteoblastic characteristics and NK-1 receptor expression of mouse MC3T3-E1 cells confirmed that serum deprivation induced apoptosis and activated classical Wnt pathway;substance P inhibited MC3T3-E1 cell apoptosis;The Wnt pathway was rapidly activated on the basis of serum-induced induction.The expression of NK-1 receptors in osteoblast-like cells is the basis of neuropeptides;the de-serum-induced expression of apoptotic gene proteins such as Bax/Bcl-2 and apoptosis protein 3,and activation of(3-cantenin expression The activation of downstream genes such as c-myc,etc.;substance P inhibited the serum-induced Bax/Bcl-2 and apoptosis protein 3 and other apoptosis-related protein expression increased;substance P can be very fast activation of ?-cantenin expression and into Nucleus was significantly faster than serum-induced rate,and its expression multiple was higher.Conclusion:During apoptosis induced by serum deprivation which can activate Wnt pathway,substance P can further rapid increase the degree of activation of the canonical Wnt pathway,thus regulating apoptosis by serum.Therefore,the results of the present study demonstrated that SP exerts a protective effect on mouse MC3T3-E1 cells confirmed that serum deprivation induced apoptosis,which related to the fast track to canonical Wnt signaling.The correlation between the "substance P-des-serum-apoptotic-Wnt pathway" axis was explained,and the related effects and significance of nerve factors in the process of fracture healing were further explained.