脐带间充质干细胞治疗雷公藤多苷诱导卵巢早衰模型小鼠的机制研究

Mechanism of human umbilical cord mesenchymal stem cells for premature ovarian failure mice model induced by Tripterygium wilfordii polyglycosides

目的研究人脐带间充质干细胞治疗雷公藤多苷诱导卵巢早衰模型小鼠的作用。方法选择SPF级BALB/c雌性小鼠58只,鼠龄7~8周龄,体质量18.0~22.2g。饲养至12周龄,连续14d灌胃给予雷公藤多苷,诱导卵巢早衰模型小鼠。按动情周期将模型小鼠随机分为模型对照组、阳性对照组和干细胞低浓度组、干细胞高浓度组,另设阴性对照组,每组12只。干细胞组小鼠尾静脉注射不同浓度的人脐带间充质干细胞,阳性对照组小鼠灌胃给予戊酸雌二醇和醋酸甲轻孕酮片。末次干预后1h、24h、7d,每组随机抽取6只小鼠,采血检测卵泡刺激素(FSH)、黄体生成激素(LH)、雌激素(E2)含量;末次干预后24h、7d,每组随机取6只小鼠取卵巢计算脏器系数并作病理观察。结果与阴性对照组比较,模型对照组末次干预后1h、24h的FSH、LH升高,末次干预后7d的E2下降,末次干预后24h、7d的卵巢系数下降;差异均有统计学意义(P<0.05或0.01)。与模型对照组比较,阳性对照组小鼠末次干预后1h的E2升高,末次干预后24h的FSH降低、卵巢系数升高,末次干预后7d的E2升高,FSH和LH降低;干细胞低浓度组24h的FSH降低,卵巢系数升高;干细胞高浓度组24h的E2升高,24h、7d的FSH降低,卵巢系数升高;差异均有统计学意义(P<0.05或0.01)。模型对照组小鼠卵巢出现萎缩、体积下降、结构混乱等病理改变,阳性对照组、干细胞低浓度组、干细胞高浓度组小鼠卵巢的病理改变均有不同程度的改善。结论人脐带间充质干细胞可使卵巢早衰小鼠血清中的FSH下降和E2升高、卵巢系数升高。故脐带间充质干细胞对卵巢早衰模型小鼠具有一定的改善作用。

Objective To evaluate human umbilical cord mesenchymal stem cells(UCMSC) in treatment of mice with premature ovarian failure(POF) induced by Tripterygium wilfordii polyglycosides.Methods The 58 SPF grade BALB/c female mice were sacrificed,which aged 7 - 8 weeks with body mass of 18.0 - 22.2 g.All of them were given Tripterygium wilfordii polyglycosides by oral administration for 14-day and induced premature ovarian failure model.Then they were randomly divided into model group,positive group,UCMSC low-dose group and UCMSC high-dose group by oestrous cycle,and 12 normal mice set as negative control group(n = 12).The UCMSC was injected into the mice of UCMSC low-dose group and high-dose group via the tail vein.Estradiol valerate and progesterone acetate were given to positive control group by oral administration.At 1- hour,24-hour,and 7-day after the last intervention,6 mice were randomly selected from each group,and the blood samples were taken to detect content of follicle stimulating hormone(FSH),luteinizing hormone(LH) and estrogen(E2);The ovary was collected 24-hour and 7-day after the final intervention.The changes of ovarian index,serum level of FSH,LH,E2 and ovarian histomorphological were observed.Results Compared with negative control group,the FSH and LH of model control group increased at 1-hour and 24-hour after final intervention,E2 decreased at 7-day after final intervention,and ovarian coefficient decreased at 24-hour and 7-day after final intervention.The difference was statistically significant(P < 0.05 or 0.01).Compared with model control group,the E2 of positive control group increased at 1-hour after final intervention,FSH decreased and ovarian coefficient increased 24-hour after final intervention.The E2 increased,FSH and LH decreased at 7-day after final intervention. The FSH of UCMSC low-dose group was decreased at 24-hour and ovarian coefficient increased.The E2 of UCMSC high-dose group was increased at 24-hour,FSH decreased at 24-hour and 7-day,the ovarian coefficient was increased, and the difference was statistically significant(P < 0.05 or 0.01).The model control group showed pathological changes such as atrophy,volume decrease,and structural disorder.The ovarian pathology of positive control group,UCMSC low-dose group and UCMSC high-dose group were all improved.Conclusion It is demonstrated that HUMSC could decrease the FSH and E2 in serum of premature ovarian failure mice,and increase the ovarian coefficient,which showed the certain improvement effect on mice with premature ovarian failure.