Effects Of Alcohol On The Expression Of Neuropeptide In Human Bone Marrow Mesenchymal Stem Cells

BackgroundNow, there are more and more Patients suffering from alcoholic avascular necrosis disease, disturbed people’s lives, but its pathogenic mechanisms are not yet clear, and so far, we haven’t found any effective ways to prevent alcohol-induced necrosis of femoral head. Its seriously affected the physical and mental health, reduced quality of life. Refers to the femoral head ischemic necrosis of femoral head blood supply side eventually led to constant death and repair of bone cells and tissue, so that changes in internal structure of the femoral head, and then collapsed, culminated in the barrier function of joints. Since the17th century academics raised the disease so far, many scholars have proposed a number of doctrines, but there is no reasonable theory to explain the pathogenic mechanisms. Bone marrow-derived mesenchymal stem cells are able to differentiate into osteogenic cells and other cells of the stem cells, studies have shown that there are neural peptides substance exist in bone marrow, which can be induced to differentiate into bone or blood vessels and play an important role in the bone tissue of metabolism process. Up to now, it has not reported that alcohol on neuropeptide expression in human bone marrow-derived mesenchymal stem cells effect. ObjectiveTo investigate the effect of alcohol on the expressions of calcitontin gene-related peptide receptor (CGRPR) mRNA, substance P receptor (SPR) mRNA, peroxisom proliferator-activeted receptor-γ (PPARy) mRNA and Runx2mRNA in human BMSCs. investigate the pathogenesis of avascular necrosis of the alcoholic.MethodsBone marrow specimens were obtained from10healthy volunteers, six men and four women, mean age57.5(54～62) years of age. Separation using density gradient centrifugation and adherent culture, remove the iliac bone marrow injected into a sealed sterile anticoagulant tube, slow Shop in percussion evenly on top of the volume of Ficoll lymphocyte separation medium, after centrifugation the middle of the slow extraction albuginealayer, after its accession to the PBS centrifuged supernatant was removed by adding complete DMEM medium and added to cell culture flasks in the incubator for the first time, medium was changed after3days, after2or3days according to the cell medium was changed, when cells are covered withbottom90%of trypsin digestion and passage, repeat the operation to the2nd generation of cells. Immunochemical staining of primary cells. The collection of2nd generation of cells were seeded and cultured in6-well culture plates, when cells covered about80percent, to four samples in each group were randomly divided into two groups, namely the experimental group and control group. The experimental group in the medium by adding0.09mol/L alcohol, normal cultured control group. RNA was extracted, and RNA purity and integrity of the test11days after the ethanol is added to the Trizol method. Reverse transcriptase synthesis of cDNA, and then by real-time fluorescent quantitative PCR (RT-of qPCR) detection and PCR products by agarose gel electrophoresis, the main observation of the experimental group and control group cells CGRP receptors, substance P receptor, PPAR-y, Runx2mRNA in the expression of. Results and conclusionThe expressions of FGF mRNA, CGRPR mRNA, SPR mRNA and Runx2mRNA in the experimental group were significantly lower than that in the control group (P<0.01), but the expression of PPARy mRNA was increased compared with control group (P<0.01). Alcohol could change the expressions of neuropeptide and depress the differentiation to osteoblasts.