Effect Of CLA, LA, PA On Proliferation And Differentiation Of Rat Preadipocyte

The experiment explored the culture condition of the rat preadipocyte in the given lab, on the basis of this investigated the effect of CLA, LA, CLA+LA, PA on proliferation and differentiation of rat preadipocyte. Primary culture of rat preadipocyte were prepared from the epididymal, inguinal and perirenal the fat pads of male normal, healthy, 15-20 days Sprague-Dawley rats.The preadipocyte grew better under the condition of 37 , 95%humidity, 5%CO2, PH 7.0-7.2, centrifuged at 1000r/min, M199medium, and 10% fetal bo vine serum, seeded at a density of 4 l04, 5 l04, /cm2. Oil red O staining was the special method to distinguish adipocyte from other cells, Gimsa and HE could determine the stage of the adiopcyte differentiation through the number of lipid drop, size and the position of the nucleolus of the staining fat cell.By counting cell, determining OD after oil red 0 staining to investigate the effect of CLA on proliferation and differentiation of rat preadipocyte. The results showed: CLA inhibited the proliferation and differentiation of rat preadipocyte through decreased the cell number and limited filling lipid. 0.05, 0.10, 0.15mmol/L CLA all could work, but the lower dose of CLA made a effect after exposed a relative long time. The dose of the 0.10, 0.15mmol/L CLA had no difference.By MTT, determinied OD after oil red O staining to investigate the effect of the CLA, LA,CLA+LA on the proliferation and differentiation of rat preadipocyte and adipocyte. CLA's function was consistent with above experiment, the LA of lower dose could decrease the cell number and depressed lipid filling; but the higher level of LA had no effect on the proliferation and differentiation of preadipocyte and adipocyte; plused LA to CLA, the effect of CLA on preadipocyte and adipocyte could partly restore, and the levels of LA had no influence on CLA's on preadipocyte and adipocyte.By counting cell, cell growth curve, cell doubling time, and the rate of the lipid filling of adipocyte to investigated the effect of PA on the proliferation and differentiation of preadipocyte. Following results were obtained: PA promoted the proliferation thoughincreasing the cell number, improved the degree of proliferation, reduced the double time of the preadipocyte; PA increased the differentiation of the adipocyte by enlarging the size of the adipocyte and improving the rate of the adipocyte; 0.10mmol/L PA had on evident effect on the proliferation and differentiation of the preadiipocyte and adipocyte. The improved influence of 0.20, 0.30mmol/L PA were significant, and the level of between 0.20 mmol/L PA and 0.30mmol/L PA presented significance of difference.