血管生成调节因子对性成熟前小鼠卵泡及其血管发育的影响
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摘要
血管生成促进因子和血管生成抑制因子的平衡控制着新生血管的生成。血管新生在卵泡的生长后期和排卵卵泡的选择上起到了至关重要的作用。血管系统的增殖与衰退伴随着卵泡的整个生长过程。因此,了解血管生成调节因子对性成熟前小鼠卵泡及其血管发育的影响是一项非常重要的研究任务,这将为卵泡血管新生的人为控制和卵巢功能障碍的治疗提供有力帮助。
本实验通过对性成熟前小鼠腹腔注射各种血管生成调节因子,如血管内皮生长因子(VEGF)、促卵泡激素(FSH)、2-甲氧基雌二醇(2-ME2)、4-羟基雌二醇(4-OHE2、FSH受体拮抗剂(FSHR antagonist)和雌二醇(E2),探讨它们对性成熟前小鼠卵巢重量、各级卵泡数目、卵泡闭锁率以及卵泡周围血管发育的影响。结果如下：
(1)FSH和VEGF可促进小鼠卵巢重量的增加,并且VEGF的这种促进作用可被FSH受体拮抗剂削弱；2-ME2、4-OHE2和2-ME2+FSH可使小鼠卵巢重量显著降低。
(2)各种处理对大、中、小卵泡数量具有不同影响。注射FSH和VEGF后大卵泡数目明显上升；注射2-ME2、4-OHE2和VEGF+FSH受体拮抗剂后大卵泡数目明显下降。VEGF可促使中卵泡数目明显上升,而FSH对中卵泡数目无显著影响；2-ME2和E2可促使中卵泡数目明显下降。注射2-ME2、VEGF后小卵泡数目明显上升,注射E2后小卵泡数目明显减少。
(3)各种处理对大、中、小卵泡闭锁率具有不同影响。抑制或促进血管发育均可造成大卵泡闭锁率明显上升；注射药品2-ME2和E2后中卵泡闭锁率明显上升,FSH能够抑制2-ME2的促闭锁作用；各种试剂对小卵泡闭锁率均无显著影响。
(4)各种处理对卵泡基膜中的血管发育具有不同影响。通过统计大卵泡基膜中的红细胞数发现,FSH和VEGF可促进大卵泡基膜中的血管发育；2-ME2和4-OHE2可抑制大卵泡基膜血管发育；VEGF促血管发育作用不依赖于FSH作用；2-ME2对血管发育的抑制作用能够被FSH解除。统计中卵泡基膜中的红细胞数发现,VEGF促进中卵泡基膜血管发育,而FSH不能,2-ME2和4-OHE2抑制卵泡血管发育,VEGF促血管发育作用依赖于FSH作用,2-ME2对血管发育的抑制作用能够被FSH解除。统计小卵泡基膜中的红细胞数发现,血管发育促进或抑制因子对小卵泡基膜中的血管发育均无显著影响。
(5)性成熟前小鼠腹腔注射各种药品后,定量测定一侧卵巢颗粒细胞VEGF、Flk-1基因表达发现,外源VEGF、FSH促进颗粒细胞目标基因表达,而且VEGF对目标基因表达的促进作用依赖于FSH、FSH的促进作用能够为2-ME2所抑制。4-OHE2和E2抑制颗粒细胞中VEGF基因的表达。
Formation of tumor new vessels is controlled by balances between angiogenic stimulators and inhibitors. Angiogenesis play essential function in the follicle development and the selection of ovulated follicles Especially in the maintain of vascular permeability. Therefore,understanding the regulation of angiogenic factor on follicular angiogenesis and development in immature mouse is a very important research task,which provides a powerful help for the artificial control of follicular angiogenesis and the treatment of ovarian dysfunction.
This study determined the effects of angiogenic factors (VEGF、FSH、2-ME2、4-OHE2、FSHR antagonist and E2) on follicle numbers, the percentage of atretic follicles and vascular development around the follicle.The results show:
(1) FSH and VEGF can increase the weight of ovary,and the effects may be reduced by FSHR antagonist.2-ME2、4-OHE2 and 2-ME2+FSH significantly decreased the ovary weight in immature mouse.
(2) Various treatments have a different impact on the number of follicles.The number of large follicles was increased after the injection of FSH and VEGF, however,it was decreased after the injection of 2-ME2、4-OHE2 and VEGF+FSHR antagonisy;The number of medium follicles was increased after the injection of VEGF, FSH had no significant effect, however,it was decreased after the injection of 2-ME2 and E2; The number of small follicles was increased after the injection of 2-ME2 and VEGF, however,it was decreased after the injection of E2.
(3) Various treatments have a different impact on the percentage of atretic follicles. There is a increase in the percentage of atretic follicles when the angiogenesis was promoted or inhibited. The percentage of atretic follicles was increased after the injection of 2-ME2 and E2,while FSH inhibited the effect of 2-ME2.
(4) Various treatments had no significant effect. Various treatments have a different impact on the angiogenesis Through statistical analyses on the number of red blood cells around the large follicles, FSH and VEGF can promote the angiogenesis,2-ME2 and 4-OHE2 can inhibit the angiogenesis. The promotion effect of VEGF does not depend on FSH.The inhibition of 2-ME2 to the angiogenesis was relieved by FSH. Through statistical analyses on the number of red blood cells around the medium follicles, VEGF can promote the angiogenesis,2-ME2 and 4-OHE2 can inhibit the angiogenesis. The promotion effect of VEGF depend on FSH. The inhibition of 2-ME2 to the angiogenesis was relieved by FSH. Through statistical analyses on the number of red blood cells around the small follicles. Various treatments had no significant effect to the angiogenesis in the small follicles.
(5) The othe side of the ovaries was added to detect the expression level of VEGF and Flk-1mRNA. According to our results:VEGF and FSH could promote the expression of the target gene, the promotion effect of VEGF depend on FSH, the promotion effect of FSH could be inhibited by 2-ME2.4-OHE2 and E2 could inhibit the expression level of VEGF.
本实验通过对性成熟前小鼠腹腔注射各种血管生成调节因子,如血管内皮生长因子(VEGF)、促卵泡激素(FSH)、2-甲氧基雌二醇(2-ME2)、4-羟基雌二醇(4-OHE2、FSH受体拮抗剂(FSHR antagonist)和雌二醇(E2),探讨它们对性成熟前小鼠卵巢重量、各级卵泡数目、卵泡闭锁率以及卵泡周围血管发育的影响。结果如下：
(1)FSH和VEGF可促进小鼠卵巢重量的增加,并且VEGF的这种促进作用可被FSH受体拮抗剂削弱；2-ME2、4-OHE2和2-ME2+FSH可使小鼠卵巢重量显著降低。
(2)各种处理对大、中、小卵泡数量具有不同影响。注射FSH和VEGF后大卵泡数目明显上升；注射2-ME2、4-OHE2和VEGF+FSH受体拮抗剂后大卵泡数目明显下降。VEGF可促使中卵泡数目明显上升,而FSH对中卵泡数目无显著影响；2-ME2和E2可促使中卵泡数目明显下降。注射2-ME2、VEGF后小卵泡数目明显上升,注射E2后小卵泡数目明显减少。
(3)各种处理对大、中、小卵泡闭锁率具有不同影响。抑制或促进血管发育均可造成大卵泡闭锁率明显上升；注射药品2-ME2和E2后中卵泡闭锁率明显上升,FSH能够抑制2-ME2的促闭锁作用；各种试剂对小卵泡闭锁率均无显著影响。
(4)各种处理对卵泡基膜中的血管发育具有不同影响。通过统计大卵泡基膜中的红细胞数发现,FSH和VEGF可促进大卵泡基膜中的血管发育；2-ME2和4-OHE2可抑制大卵泡基膜血管发育；VEGF促血管发育作用不依赖于FSH作用；2-ME2对血管发育的抑制作用能够被FSH解除。统计中卵泡基膜中的红细胞数发现,VEGF促进中卵泡基膜血管发育,而FSH不能,2-ME2和4-OHE2抑制卵泡血管发育,VEGF促血管发育作用依赖于FSH作用,2-ME2对血管发育的抑制作用能够被FSH解除。统计小卵泡基膜中的红细胞数发现,血管发育促进或抑制因子对小卵泡基膜中的血管发育均无显著影响。
(5)性成熟前小鼠腹腔注射各种药品后,定量测定一侧卵巢颗粒细胞VEGF、Flk-1基因表达发现,外源VEGF、FSH促进颗粒细胞目标基因表达,而且VEGF对目标基因表达的促进作用依赖于FSH、FSH的促进作用能够为2-ME2所抑制。4-OHE2和E2抑制颗粒细胞中VEGF基因的表达。
Formation of tumor new vessels is controlled by balances between angiogenic stimulators and inhibitors. Angiogenesis play essential function in the follicle development and the selection of ovulated follicles Especially in the maintain of vascular permeability. Therefore,understanding the regulation of angiogenic factor on follicular angiogenesis and development in immature mouse is a very important research task,which provides a powerful help for the artificial control of follicular angiogenesis and the treatment of ovarian dysfunction.
This study determined the effects of angiogenic factors (VEGF、FSH、2-ME2、4-OHE2、FSHR antagonist and E2) on follicle numbers, the percentage of atretic follicles and vascular development around the follicle.The results show:
(1) FSH and VEGF can increase the weight of ovary,and the effects may be reduced by FSHR antagonist.2-ME2、4-OHE2 and 2-ME2+FSH significantly decreased the ovary weight in immature mouse.
(2) Various treatments have a different impact on the number of follicles.The number of large follicles was increased after the injection of FSH and VEGF, however,it was decreased after the injection of 2-ME2、4-OHE2 and VEGF+FSHR antagonisy;The number of medium follicles was increased after the injection of VEGF, FSH had no significant effect, however,it was decreased after the injection of 2-ME2 and E2; The number of small follicles was increased after the injection of 2-ME2 and VEGF, however,it was decreased after the injection of E2.
(3) Various treatments have a different impact on the percentage of atretic follicles. There is a increase in the percentage of atretic follicles when the angiogenesis was promoted or inhibited. The percentage of atretic follicles was increased after the injection of 2-ME2 and E2,while FSH inhibited the effect of 2-ME2.
(4) Various treatments had no significant effect. Various treatments have a different impact on the angiogenesis Through statistical analyses on the number of red blood cells around the large follicles, FSH and VEGF can promote the angiogenesis,2-ME2 and 4-OHE2 can inhibit the angiogenesis. The promotion effect of VEGF does not depend on FSH.The inhibition of 2-ME2 to the angiogenesis was relieved by FSH. Through statistical analyses on the number of red blood cells around the medium follicles, VEGF can promote the angiogenesis,2-ME2 and 4-OHE2 can inhibit the angiogenesis. The promotion effect of VEGF depend on FSH. The inhibition of 2-ME2 to the angiogenesis was relieved by FSH. Through statistical analyses on the number of red blood cells around the small follicles. Various treatments had no significant effect to the angiogenesis in the small follicles.
(5) The othe side of the ovaries was added to detect the expression level of VEGF and Flk-1mRNA. According to our results:VEGF and FSH could promote the expression of the target gene, the promotion effect of VEGF depend on FSH, the promotion effect of FSH could be inhibited by 2-ME2.4-OHE2 and E2 could inhibit the expression level of VEGF.
引文
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