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目的观察小鼠卵泡发育过程中缝隙连接蛋白43(Cx43)和黄体生成素受体(LHR)在小鼠卵巢中的表达以及黄体生成素(LH)对卵巢Cx43表达的影响,为这两种分子与卵泡发育的关系提供实验依据。方法选择出生后1d、4d、7d、2周、3周、4周、6周、8周龄雌性C57小鼠共8组,每组10只,取卵巢。HE染色观察卵巢形态、卵泡发育并测量颗粒细胞体积分数。免疫组织化学和Western blotting观察卵巢组织中Cx43及LHR的表达,Western blotting检测不同浓度黄体生成素(LH)孵育后对卵巢Cx43表达的影响。结果 HE染色显示,出生1d小鼠卵巢中见原始卵泡,出生7d卵巢内出现初级卵泡,2周时出现窦前卵泡和少量窦卵泡,3周、4周时,出现较多大的窦卵泡,6~8周时出现黄体。卵泡颗粒细胞体积分数在2周后的卵巢中明显增加。Cx43表达在各阶段卵泡的颗粒细胞胞膜上,LHR表达在卵泡膜细胞、颗粒细胞及卵母细胞胞质中。Cx43和LHR出生后2周卵巢中的表达开始明显增加,并随卵泡发育表达逐渐增强,在3周、4周、6周和8周维持相对较高水平。LH体外干预可增加卵巢组织Cx43的表达。结论 Cx43和LHR在卵泡发育中发挥重要作用,LH可能通过LHR促进卵巢卵泡颗粒细胞Cx43的表达。
Objective To observe the expression of connexin 43 (Cx43) and luteinizing hormone receptor (LHR) in mouse ovary and the effect of luteinizing hormone (LH) on the expression of ovary Cx43 during follicular development in mice. And the relationship between follicular development to provide experimental evidence. Methods Female C57 mice of 1d, 4d, 7d, 2week, 3week, 4week, 6weeks and 8weeks after birth were selected and divided into 8 groups, with 10 in each group and ovariectomized. Ovarian morphology, follicular development and granulosa cell volume fraction were observed by HE staining. Immunohistochemistry and Western blotting were used to observe the expression of Cx43 and LHR in ovarian tissue. Western blotting was used to detect the effect of different concentrations of luteinizing hormone (LH) on the expression of Cx43 in ovary. Results HE staining showed that primordial follicles were found in the ovaries of the first day after birth, primary follicles were found in the ovaries 7 days after birth, preantral follicles and antral follicles were found at 2 weeks, and larger antral follicles appeared at 3 weeks and 4 weeks. 6 ~ 8 weeks when the body appears. Follicular granulosa cell volume fraction increased significantly after 2 weeks of ovary. Cx43 is expressed on the granulosa cell membrane of follicles at various stages, and LHR is expressed in the cytoplasm of theca cells, granulosa cells and oocytes. The expression of ovary in Cx43 and LHR 2 weeks after birth significantly increased, and gradually increased with the follicular development, and remained relatively high at 3 weeks, 4 weeks, 6 weeks and 8 weeks. LH in vitro can increase the expression of Cx43 in ovarian tissue. Conclusion Cx43 and LHR play an important role in follicular development. LH may promote the expression of Cx43 in ovarian follicle granulosa cells through LHR.