hUCMSCs移植提高围绝经期大鼠血清雌激素水平的持续时间及对卵巢P450arom表达的影响

目的:探讨人脐带间充质干细胞(h UCMSCs)移植提高围绝经期大鼠血清雌激素(E2)水平的持续时间及其对卵巢细胞色素P450芳香化酶(P450arom)表达的影响。方法:阴道脱落细胞涂片法筛选出30只SPF级12~14月龄的围绝经期SD大鼠,随机分为A、B组;同时筛选出15只SPF级3~5月龄具有生育能力的青年雌性SD大鼠,作为C组。B组:尾静脉移植浓度为1×10~6cells/ml的h UCMSCs单细胞悬液1ml,隔日1次,共两次;A组:与B组在相同时间相同方式移植等量PBS液;C组:不予处理。于移植前所有大鼠经眼眶采血检测血清E2水平,并于末次移植后14d、21d、28d每组随机选取5只大鼠采血并取双侧卵巢组织,ELISA法检测血清E_2水平、免疫组化及Western blot法检测卵巢P450arom表达水平。结果:末次移植后14d、21d、28d,B组大鼠血清E_2水平、卵巢P450arom表达与移植前及A组相比均明显升高,差异有统计学意义(P0.05),B组3个时间段的E_2水平、P450arom表达无明显差异(P0.05)。结论:h UCMSCs移植提高围绝经期大鼠血清E_2水平可至少持续至移植后28d;h UCMSCs移植可促进围绝经期大鼠卵巢细胞色素P450芳香化酶表达,是其提高E_2水平的可能途径。

The effect of hUCMSCs transplantation on the duration of serum estrogen level and ovarian P450arom expression of perimenopausal rats

Objective:To investigate the effect of human umbilical cord mesenchymal stem cells ( hUCMSCs) transplantation on the duration of serum estrogen ( E2 ) level and its effect on the expression of aromatase cytochrome P450 (P450arom) in the perimenopausal rats. Methods:The 30 natural aging female Sprague-Dawley rats (12~14 months old) of SPF class were selected through vaginal smears, which were randomly divided into A and B groups. We selected 15 young female SD rats (3~5 months old) with fertility as C group at the same time. Group B was transplanted 1ml hUCMSCs suspension at the concentration of 1 × 106 cells/ml through tail vein. After 48 hours,group B was again injected with 1ml hUCMSCs suspension. Group A was injected with 1ml PBS per rat via the tail vein at the same time. Group C were ac-cepted no any treatments. Before transplantation,1ml of blood was gathered from each rat's or-bital. At 14,21 and 28 days after the final cell transplantation,the five rats from each group were selected randomly and euthanized. Blood was taken from orbital and ovaries were removed for ELISA analysis of E2 levels, immunohistochemistry and Western blot detection of ovarian P450arom expression. Results:Serum level of E2 and ovarian expression of P450arom were in-creased in group B at time-points of 14 ,21 ,28 days after transplantation ( P<0 . 05 ) . The level of E2 and ovarian expression of P450arom had no significant difference in group B at three time-points(P>0. 05). Conclusion:hUCMSCs improve serum E2 level of perimenopausal rats at least 28 days after transplantation. HUCMSCs transplantation may improve E2 level of perimeno-pausal rats via promoting ovarian P450arom expression.