The Effect Of Psoralen On Proliferation Capacity, Differentiation To Adipocyte Of Marrow Stromal Cells From Rats

Objective:To observe the biologic characteristic of 3-month SD rats and to observe the effect onproliferation capacity, differentiation to adipocyte of SD rat's marrow stromal cellcultured by various concentrations of Psoralen and confirm the best concentrationand duration of operation. To investigate the expression of PPARγmRNA in inducedsystem with Psoralen in molecular biology leval. The study may provide a betterunderstanding of the mechanisms involved in the differentiation of 3-month SD rats.Methods:1. To culture MSCs of 3-month SD rats successful. 10% calf serum and DMEM-F12can promote the growth of MSCs in vitro; MSCs from SD rats were purified by usingDifferential Attachment method, then identified according to morphology, FCM andinduced-differentiation potentiality;2. Measuring the cell growth curve of MSCs by means of MTT;3. Different concentrations of Psoralen characterized as 5 concentrations were addedto rat MSCs. Proliferation between Psoralen groups (different concentrations) andblank comparison group were detected by means of MTT;4. Different concentrations of Psoralen characterized as 5 concentrations were addedto rat MSCs which is divided into the Psoralen+induced-differential-to-adipocytegroups (different concentrations), the blank comparison group, and theinduced-differential-to-adipocyte group. To detect the effect on differentiation toadipocyte of SD rat's MSCs by use of oil-red O staining, PNPP and enzyme method.5.RT-PCR method was used to detect the expression of PPARγ. Grouping was same asabove. Results:1. Primary cultured MSCs adhered to plastic surface within 24 hours and reachedconfluence within 10-15days. Under conditioned inducement, MSCs were successfullydifferentiated into osteoblast, adipocyte;2. MSCs were confirmed that there was expression of CD29 and CD44, and noexpression of CD34 and CD45 on the surface of MSCs.3. The Psoralen have the proliferation effects on MSCs in SD rats. The best promotionof proliferation of MSCs was achieved when the concentration of Psoralen was 1μmol/L and 5μmol/L.(P＜0.05).4. Used oil-red O staining counting method, we found different concentration ofPsoralen such as5μmol/L,10μmol/L,15μmol/L and 20μmol/L in the inductionprocess can inhibit the formation of adipocyte, (P＜0.05), Differences are withstatistical significance.5. We found Psoralen in the induction process has increased the secretion of ALP andinhibit the secretion of TG.. ALP content : induced-differential-to-adipocyte group＜Psoralen+induced-differential-to-adipocyte group＜blank-comparison-group,(P＜0.05); TG content: induced-differential-to-adipocyte group＞Psoralen+induced-differential-to-adipocyte group＞blank-comparison group.6. Psoralen has downward effect on PPARγexpression in induction system. PPARγexpression (overall mean): induced-differential-to-adipocyte group＞Psoraleninduced-differential-to-adipocyte group＞blank-comparison group(P＜0.05).Conclusion:1. MSCs can be cultured and propagated in vitro;2. Some concentrations of Psoralen as 1μmol/L and 5μmol/L have theproliferation effects on MSCs in SD rats.3. Different concentrations of Psoralen characterized as 5～20μmol/L could inhibitadipocyte differentiation, lower TG content, increase the ALP content in MSCs andpromot MSCs into osteoblast.4. 5μmol/L concentration of Psoralen can reduce the expression of PPARγmRNA,...