| Objective:In order to investigate the curative effects of 3T3-L1 preadipocytes in polycystic ovary syndrome(PCOS) mice, green fluorescent protein(GFP) derived 3T3-L1 preadipocytes were transplanted and their homing was tracked in a mice model of PCOS. And then, the numbers, sizes and maturity of follicles in ovary of mouses were observed to speculate the ovulation situation. Effects of hyperinsulinism and high-testosterone on adipose-derived stem cells(ADSCs) isolated from human beings and cultured in vito were reserched. In this way, we hope to find a new view and method to treat PCOS clinically. Methods:1. 23-day-old female C57/BL6 mices(SPF) were used, which were raised with free water and diet for 1 week to adapt to light cycle. The experimental animals were divided into blank control group(Wild- type, WT): normal untreated mice; PCOS group: PCOS model without any treatment; Intravenous(IV) group: 3T3-L1 preadipocyte suspension was injected via caudal vein to PCOS model.2. GFP-labeled 3T3-L1 preadipocytes were obtained via lentiviral transfection. 0.2 ml of cell suspension(1×106/ml) was injected via caudal vein, and the process was repeated on the next day. Frozen section of mice ovary was taken 1 week and 1 month after the implantation, and 3T3-L1 preadipocyte homing to the ovary under fluorescent microscope was observed.3. 1 week or 1 month following implantation of ADSCs, ovarian tissues of mice were excised. Ovarian tissues of mice were fixed, embedded and sliced, and histopathologic morphology was used to observe the morphology and structure of ovary and number of follicles after HE staining.4. Subcutaneus adipose tissues were taken from healthy pregnant women via cesarean surgery. Adipose stem cells were isolated, cultured and identified and treated with high concentration(10-4 mol/L), medium concentration(10-5 mol/L) and low concentration(10-6 mol/L) of testosterone and insulin. Using Western Blot to detect expressions of PPARγ and a P2 in adipose stem cells.5. SPSS17.0 analysis software was used to carry out a statistical processing of experimental data. The expressions of PPARγ and a P2 and the number of preovulatory follicles of mice were shown via(). One-way ANOVA was used for variance among groups, with P<0.05 as the standard of statistically significant differences.Results:1.3T3-L1 preadipocytes with GFP(+) gene could be stably carried by lentiviral transfection, and could be tracked in animals.2.Homing of 3T3-L1 preadipocytes could be observed in the ovaries of PCOS mice models after being transplanted with 3T3-L1 preadipocytes.3.Mature follicles were decreased in ovaries of PCOS mice, and the number of preovulatory follicles were increased after being transplanted with 3T3-L1 preadipocytes in 1 week and 1 month; Comparison between the control group and 1 week group, the difference was considered to have no statistical significance(P>0.05); but the difference between the control group and 1 month group was considered to have statistical significance(P<0.05).4.ADSCs of human in virtro culture system was successfully established via digestion and culture of human adipose tissue.5.Hyperinsulinism and high-testosterone could influence the function of adipose stem cells system in vitro; it can be obtained that there is significant correlation between increase of drug concentration and decrease of PPARγ and a P2 expressions; there were statistically significant differences among high-, mediumand low- concentration groups and blank control groups(P<0.05).Conclusions:1. 3T3-L1 preadipocytes can improve the ovarian function of PCOS mice, and may have therapeutic effect of PCOS.2. High insulin and high-testosterone can affect the function of adipose endocrine at the level of adipose stem cells. |
PDF Full Text Request