Effect Of EMO And RH On The Proliferation And Differentiation Of Rat Preadipocytes

In the research, stromal vascular cells( SV cells) from SD rat fat mass was adopted to separate preadipocytes, and the condition of culture media was studied that can make preadipocytes reach better states of proliferation and differentiation. On the base, the methods of oil red straining, morphologic observation, MTT spectrophotometry and flow cytometry(FCM) was applied to determine the effects of emodin and rhein on the proliferation, differentiation and apoptosis of rat preadipocytes. At the same time, the mechanism of its effect was discussed principially. This results might act as the basis that emodin and rhein could be used as medicament or additive to regulate and control the fat aggradation in human or animal body. It perhaps is hopeful that the time-effect relation of Chinese traditional medicine could be improved, and exert its multilayer and multi-target pharmacology function. This research includes following several aspects: 1 To study the proper culture media that can make the rat preadipocytes reach better state of proliferation and differentiation . L-Gln was added to the four kinds ofculture media with different initial nutritial ingrediants. The proliferation and differentiation of preadipocytes were determined by MTT spectrophotometry and Oil Red O staining, respectively. The results showing that the addition of 0 mmol/L L-Gln in fresh culture media and 0.685 mmol/L L-Gln in the culture media that having been stored at 4℃for 20d benefited for proliforation and differentiation of the preadipocytes. When culture media was stored at 4℃for 90d, it can not make the preadipocytes developing formally, even if L-Gln was added. Once more, M199 and FCS(fetal calf serum) cannot be stored at 4oC beyond 90d if they are opened. 2 To study the effect of EMO(emodin) and RH(rhein) on the differentiation of rat preadipocytes. Accumulation of TG(triglyceride) in the adipocytes was determined by oil red O staining, and the cell percentage and degree of lipid accumulation were determined by morphology observation. The results showing that EMO and RH inhibited preadipocytes differentiation in a dose and time dependent manner, and the suppression function of EMO was stronger than RH. 3 To study the effect of EMO(emodin) and RH(rhein) on proliferation of rat preadipocytes. MTT spectrophotometry and cell cycle was assayed by FCM(flow cytometry) was adopted. The Results showing that In the effective range of dose, EMO and RH inhibited the proliferation of preadipocytes in a dose and time dependent manner; the suppression function of EMO and RH were performed in different phase of cell cycle, and both they could induce apoptosis of preadipocytes. This research conclude following: Suitable supplement of L-Gln could make preadipocytes approach better states of proliferation and differention; Both EMO and RH could inhibit proliferation and differention of the preadipocytes in a dose and time dependent manner, but perhaps their suppression effect was in different mechamism. EMO and RH should have a potential to serve as a fat-reducing drug.