The Effect Of Ethanol On The MRNA Expression Of Neuropeptide Receptor And Apoptosis In Human Bone Marrow Mesenchymal Stem Cells

Background:The ethanol-induced necrosis of femoral head cases beacame more and more in in recent years. But the accurate etiopathogenesis of ONFH still unclear. Pervious studies showed that there is hBMSCs exist in human bone marrow which could differentiate into osteoblast, adipocyte,chondroblast, or neurocyte, et al. This different-iation is influenced by a variety of neurotransmitters. And some existing in the bone marrow of neurotransmitters can promote osteogenesis and vascular function to maintain normal bone metabolism.Apoptosis is a natural physiological process, due to internal and external environment stimulate cells themselves actively and orderly manner of death which process involves accurate gene transcription and protein synthesis.The caspase is the core element of the apoptosis pathway. The active measurement of caspase-3has become an important indicator apoptosis related research as apoptosis protein important effect.The Bcl-2can inhibit the role of apoptosis as a kind of important genes regulate cell apoptosis.Objective:To observe the effect of neuropeptide or its receptor and apoptosis mRNA expre-ssion in human bone marrow mesenclymal stem cells (hBMSCs) under the ethanol condition.Methods:The hBMSCs were cultivated and isolated by adherence and density gradient centrifugation and identifided by flow cytometry. The3rd passage of hBMSCs was randomly divided into two groups. In the experimental group, the cells were treated with0.09mol/L ethanol. In the control group, the cells were normally treated without ethanol.The mRNA expressions of Peroxisom proliferator-activeted receptor-y (PPAR-y),Calcitonin gene-related peptide receptor(CGRPR),Runt-related transcri-ption factor(Runx2),B cell lymphoma-2(bcl-2) and Cysteine aspartic acid specific protease-3(caspase-3) of hBMSCs were detected by using real-time quantitative reverse transcription-polymerase chain reaction(RT-qPCR) on days11.Results:Adherent cells showed that fibroblast-figure spindle or shape shaped swiral, morphology or radial growth on day11.The positive rate of CD44and CD29was (94.2±0.5)%and (92.3±0.7)%,while CD45and CD34was (5.6±0.5)%and (2.7±0.8)%.These accord showed that obtained adherent cells are hBMSCs.The expressions of CGRPR、Runx2and bcl-2mRNA in the experimental group were significantly lower than that in the control group, CGRPR (1.90E-01±4.12E-02 vs3.27E-01±8.24E-02, t=6.66, p<0.05)> Runx2(1.28E-01±3.18E-02vs2.50E-01±5.33E-02, t=8.80, p<0.05)> bcl-2(1.65E-01±3.98E-02vs2.95E-01±6.78E-02, t=7.35, p<0.05), but the expression of PPAR-γ and caspase-3mRNA increased significantly, Caspase-3(5.27E-01±1.26E-01vs3.17E-01±7.46E-02, t=6.39, p<0.05), PPAR-γ (2.90E-01±6.74E-02vs1.15E-01±3.36E-02, t=10.38, p<0.05)Conclusion:The ethanol can stimulate upregulation of the expression of apoptosis gene and adipogenic gene in hBMSCs and downregulation of the expression of neuropeptide factors and anti-apoptosis gene, and then promote the apoptosis and adipogeneic differetiantion of the cells and inhibit their osteogenic differentiantion, which may be related to the pathogenesis of the ONFH.