The Influence Of Tumor Necrosis Factor-alpha For The Chemerin Signaling Pathway Over The Cellular Level In Adipocyte Differentiation Process

With the improvement of living standards, dietary changes, life and workingpressure increased, obesity becomes more and more widespread and growing problem.Obesity is characterized by too much adipose tissue, it affects not only the visualappearance, but be a cause of inflammation, immune regulation and anneuro-humoralregulation changes. It could induce high blood pressure, heart disease, atherosclerosisand other cardiovascular diseases. It seriously endangers people’s health and life. Nowmore and more people concern about harms of obesity, so the current research onobesity in depth is done to find reasonable and healthy way to inhibit and treatobesity.As an important secretion organs, adipose tissue can secrete a lot of their fatfactors playing an important role to itself, such as TNF-α、chemerin and adiponectin,etc. This paper studys on TNF-α and chemerin/CMKLR1signal transductionmechanisms in preadipocyte differentiation process. The aim is to clear therelationship between TNF-α and chemerin/CMKLR1signaling pathway in theadipocyte differentiation process. An in-depth study about it is hoped for the treatmentof obesity.1) By extracting genomic DNA in mice, then after accurate Primer Design and PCRreactions, the experimental mouse models are established for fast reliable andsufficient access to experimental mice.2) Getting white adipose tissue in groin of mice in vitro, to get the mouse primarypreadipocytes. Using different methods to induce differentiation of preadipocytes ofmice, Determine to use Dorothy B. Hausman’s method as the method of inducingdifferentiation of preadipocytes into adipocytes. Found by oil red o stain, use thismethod for simply inducing differentiation of preadipocytes into adipocytes, theadipogenic induction rate of up to80%percent, thus successfully set up before theprimary mouse cells of adipogenic differentiation model, for material security infollow-up experiments.3) The experiment, the process of adipogenic differentiation iconic factorexpression was detected by immunoblotting. Detected, the expression of adiponectinand PPARγ were higher with the increase of maturity of the fat cells, which isconsistent with the current study results obtained and further prove the reliability ofthe experimental.4) During differentiation of WT mice in primary preadipocytes, overexpress TNF-αby adding TNF-α, Through comparing in the experiments, Final concentration ofTNF-α is100ng/ml. The concentrations of TNF-α could significantly inhibitadipocyte differentiation.5) Through the extraction of the cells of different induction period mRNA of TNF-α-/-mice、WT mice and WT mice by adding100ng/ml TNF-α-/-. and thedetection of the expression of key molecular chemerin and CMKLR1in the chemerinsignal path, we found in the process of adipocyte differentiation, chemerin andCMKLR1expression gradually increased; in the same time, chemerin and CMKLR1of TNF-α deficient cells expressing higher than normal expression group TNF-α, inthe same time, TNF-α normal expression group is higher than the overexpression ofTNF-α group.This proved that TNF-α can suppress the expression of chemerin and CMKLR1,thereby inhibiting the signaling pathway of chemerin. However, the manner in whichthey inhibit the expression of chemerin signaling pathway by acting directly orindirectly effect、are there any other relevant molecules involved in is unclear andneeds to do in-depth research.