Effect And Mechanism Of Prevention Of Obesity On Pre-adipocyte Proliferation And Differentiation Of Massage Dynamic Mechanical Stimulation

Objective:From study the dynamic mechanical stimulation of former fat cells proliferation, peroxidase (pod) and the proliferation of activated receptors (peroxisome proliferators activated receptor PPAR), transcription factors, CCAAT enhancer binding protein (CCAAT enhancer binding protein C/EBP) mRNA and adiponectin receptor (AdipodRl, AdipodR2) research on the effects of expression, For the massage massage provide theoretical and experimental basis for prevention and treatment of simple obesity.Methods:Methods:l,We used the28-day-old male Sprague Dawley Rat as experimental material.sterile acquisition the around of epididymis’adipose tissue,isolation and culture mature adipocity by using of enzyme digestion. Through morphology observe,inspection mature adipocity indicator marker gene adiponectin by Oil Red O staining method and RT-PCR method to explore primary cell authentication, and with constant temperature culture device to load dynamic mechanical for cell.2,Before the fat cells and randomly divided into five groups, Or group,60r,90r,120r,150r,180r group. After24h training synchronization, according to the corresponding category loading force different intensity of stimulation, stimulation of2times a day, each time30min, the interval time of12h and3days in a row, a total of stimuli6times. After the stimulation, determined by MTT test cell growth dynamic method is used to detect the dynamic change of each cell.3,Repetition of fat cells before the group strength stimuli in method2, after the stimulus, collecting cells extracted total RNA, using RT-PCR semi-quantitative analysis AdipodR1AdipodR2, PPAR, technology and C/EBP changes in gene expression; Extraction of total protein, cells using western blot analysis, AdipoRl and AdipoR2PPAR, technology and C/EBP protein expression changes.Results:1,The original generation of SD rat fat cells before success. Oil red O staining results were positive. The cells can be successfully induced into mature fat cells. Provides a platform for the whole experiment.2, Will be fat cells before and after every end of the stimulus with the groups of light absorption value determined by MTT method and through the stimulation of6times, draw the groups of cells after different efforts to stimulate growth curve, observation is the cell vitality, increase with the gradient mechanical stimulus as a result, before the fat cells proliferation was suppressed, and the stimulus and proliferation inhibition rate are positively related. Through statistical analysis concluded that the mechanical stimulus, the greater the massage before the lower the fat cell vitality, proliferation inhibition rate is higher.RT-PCR Semi-quantitative analysis of various speed between group C/EBP, PPAR, AdipodRl, AdipodR2, changes in the level of gene expression,1C/EBP gene expression:the experimental group (group60r/min and90r/min,120r/min and150r/min and80r/min group)0r group compared with control group, the relative expression level C/EBP were lower than0r group, with significant difference (P<0.05), showed a trend of lower; Groups of two contrast also exists significant differences.(2) the PPAR gene expression:compared with0r group, the experimental group the relative expression level of PPAR is on the decline, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.(3) Adipod Rl gene expression:compared with0r group, experimental group Adipod Rl the relative expression level of a downward trend, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.(4) AdipodR2gene expression:compared with0r group, experimental group Adipod R2the relative expression level of a downward trend, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.Western Blot Western Blot test analysis and speed between group C/EBP, PPAR, AdipodRl, AdipodR2, the change of protein expression levels,(1) C/EBP protein expression:the experimental group (group60r/min and90r/min,120r/min and150r/min and80r/min group)0r group compared with control group, the expression of C/EBP levels were lower than0r group, with significant difference (P<0.05), showed a trend of lower; Groups of two contrast also exists significant differences.(2) the PPAR protein expression:compared with0r group, the experimental group of PPAR expression level is on the decline, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.(3) Adipod R1protein expression:compared with0r group, experimental group Adipod R1the expression level of a downward trend, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.(4) AdipodR2protein expression: compared with0r group, experimental group Adipod R2the expression level of a downward trend, there were significant differences (P<0.05); Groups of two contrast also exists significant differences.Conclusion:1, Under existing experimental conditions, through the maximum force environment simulation of mature fat cells in the body, and successfully established the mechanical mechanism of the model simulation massage, for simple obesity disease prevention and control were discussed from Angle of mature fat cells laid a foundation.2, In vitro simulated massage dynamic mechanics before stimulation inhibits fat cells proliferation. Dynamic mechanical stimulation by inhibiting fat cells proliferation before massage can prevention and treatment of simple obesity disease is one of the important mechanisms.3, Formerly fat cells to differentiate into mature fat cells and energy storage is the only way to obesity occurs. Experiments show that fat cells can massage before mechanical stimulation, thereby weakening PPAR and C/EBP, Adipod R1, Adipod R2gene and protein expression of mechanical stimulation can effectively prove the massage massage before participating in adipocyte differentiation and lipid metabolism, energy metabolism, and further proved that in vitro massage mechanics under the interference of former fat cells into mature adipocyte differentiation process has been effectively suppressed, shows that fat cells before can actively participate in and adapt to changes in the growth environment and mechanical environment this may be the massage massage one of the important mechanisms of prevention and treatment of obesity.