两种间充质干细胞干预化疗损伤卵巢功能分析与生物力学研究

发布时间：2018-12-11 08:06

【摘要】：本实验通过复制化疗损伤卵巢动物模型,利用分离培养出的人羊膜间充质干细胞(amniotic mesenchymalstem cells,AMSCs)与人脐带间充质干细胞(umbilical cord mesenchymal stem cells,UC-MSCs)进行干预,取对照组、模型组、AMSCs干预组、UC-MSCs干预组四组卵巢,以光学显微镜进行组织形态学观察、卵泡计数等相关分析。取各组大鼠不同时间点(干预后14天、30天、60天、90天)的血清测量E2、FSH、LH、VEGF、IGF-1的含量;取各组大鼠干预后30天、90天时的卵巢组织试样,以电子万能试验机进行拉伸实验、应力松弛实验、蠕变实验。对AMSCs干预组、UC-MSCs干预组大鼠干预结束后15d,按照雌雄比例2:1饲养,均饲养6个月,观察两组大鼠生育能力的恢复情况,观察生育的幼鼠生长发育情况。结果表明,AMSCs干预组、UC-MSCs干预组血清中各时间点E2、VEGF、IGF-1测定值均大于化疗损伤卵巢动物模型组;FSH、LH测定值均小于模型组,差异显著(p0.05)。AMSCs干预组、UC-MSCs干预组各时间点始基卵泡、窦前卵泡、窦状卵泡及排卵前卵泡数均高于化疗损伤卵巢动物模型组,差异显著(p0.05)。AMSCs干预组、UC-MSCs干预组拉伸最大应力、最大应变大于化疗损伤卵巢动物模型组,差异显著(p0.05)。AMSCs干预组、UC-MSCs干预组大鼠血清中性激素水平、分子生物学指标具有一定的恢复,卵巢的组织形态、卵泡的数量、卵巢的弹性和韧性也得到一定的恢复,并且部分大鼠恢复了生育能力。创新点:以往对化疗损伤卵巢动物模型药物干预治疗效果的研究多以分子生物学、组织形态学观察等进行分析。本文除了从上述方面进行分析研究,还加入了卵巢的拉伸力学特性、应力松弛、蠕变特性等生物力学研究,进一步探讨化疗损伤卵巢动物模型以AMSCs、UC-MSCs干预治疗的效果,为临床提供了理论基础和实验依据。
[Abstract]:In this experiment, the ovarian model was induced by chemotherapy, and the cultured human amniotic mesenchymal stem cells (amniotic mesenchymalstem cells,AMSCs) and human umbilical cord mesenchymal stem cells (umbilical cord mesenchymal stem cells,UC-MSCs) were used to intervene. AMSCs intervention group, UC-MSCs intervention group, four groups of ovary, optical microscope for histomorphological observation, follicle count and other related analysis. The serum of each group of rats at different time points (14 days, 30 days, 60 days, 90 days after the intervention) was taken to measure the content of E2FSHLHV EGFF-1; The ovarian tissue samples of each group were taken 30 days and 90 days after intervention, and the tensile test, stress relaxation test and creep test were carried out with the electronic universal testing machine. The rats in AMSCs intervention group and UC-MSCs intervention group were fed at 2:1 for 6 months according to the ratio of male and female at the end of the intervention. The recovery of fertility and the growth and development of the young rats in the two groups were observed. The results showed that the levels of E2VEGFIGF-1 in serum of AMSCs intervention group and UC-MSCs intervention group were higher than those of ovarian model group after chemotherapy. The number of primary follicles, preantral follicles, antral follicles and antral follicles in UC-MSCs intervention group were higher than those in the model group after chemotherapy. The difference was significant (p0. 05). AMSCs intervention group, UC-MSCs intervention group, the maximal tensile stress and maximum strain were higher than that of chemotherapy induced ovarian injury model group, the difference was significant (p0. 05). AMSCs intervention group, p0. 05). AMSCs intervention group). In UC-MSCs intervention group, the levels of serum sex hormones and molecular biological indexes recovered to a certain extent, and some of the rats recovered their fertility, such as the morphology of ovary, the number of follicles, the elasticity and toughness of ovary. Innovation: the previous studies on the therapeutic effect of drug intervention on ovarian model with chemotherapy injury were analyzed by molecular biology and histomorphology observation. In addition to the above aspects, the biomechanical studies on the tensile mechanical properties, stress relaxation and creep properties of the ovary were added to further explore the effect of AMSCs,UC-MSCs intervention in the treatment of the ovarian model with chemotherapy injury. It provides theoretical basis and experimental basis for clinical practice.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R711.75

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