HSP27在黄体期牦牛主要生殖器官中表达差异的研究

Study on expression difference of HSP27 in the main reproductive organs of yak at the luteal phase

为了探讨正常生理条件下牦牛热休克蛋白27(HSP27)在黄体期牦牛主要生殖器官中的表达差异,采用实时荧光定量PCR(RT-qPCR)检测了HSP27基因在黄体期牦牛卵巢、输卵管和子宫中的相对表达量,用Western-blot和免疫组织化学SP法研究了HSP27蛋白在黄体期牦牛卵巢、输卵管和子宫中的表达量和组织内分布。RT-qPCR分析表明,HSP27基因在黄体期牦牛卵巢、输卵管和子宫中均有表达,其中卵巢中的表达量分别是输卵管和子宫组织的1.03倍和4.63倍。免疫组织化学结果表明,牦牛卵泡膜内层、颗粒层、输卵管黏膜上皮、子宫内膜上皮和子宫腺中均有HSP27阳性表达。Western-blot和免疫组织化学结果表明,HSP27蛋白在黄体期牦牛各主要生殖器官组织中表达差异极显著(P<0.01),其中在卵巢中表达量最高,在子宫中表达量最低。本试验结果表明,HSP27在黄体期牦牛的各个主要生殖器官中存在表达差异,这为进一步研究HSP27在哺乳动物生殖过程中发挥的作用提供了理论依据。

In order to research the expression differences of heat shock protein 27（HSP27） in the main reproductive organs of yak at the luteal phase, the relative expression of HSP27 gene in yak ovaries,oviducts and uteri tissues was measured by using real-time quantitative PCR（RT-qPCR）. The expression and distribution of HSP27 in yak ovaries,oviducts and uteri were examined by Western-blot and immunohisto- chemistry SP method. RT-qPCR analysis manifested that HSP27 gene was all expressed in the ovaries,oviducts and uteri,the expression quantity of ovaries was 1.03 times and 4.63 times of oviducts and uteri tissue,respectively. Immunohistochemistry result indicated that HSP27 was positively expressed in yak theca folliculi interna, stratum granulosum, oviduct epithelium, endometrium epithelium and uterine glands, respectively. Western-blot and immunohistochemistry results also demonstrated that the expression of HSP27 in the main reproductive organs of yak at the luteal phase was extremely remarkably different（P〈 0.01） ,of which the expression quantity in ovaries was the highest,while in uteri was the lowest. The experimental results manifested that the expression of HSP27 in main reproductive organs of yak at the luteal phase have differences. Anyway, the study provides the theoretical foundation for further investigation of HSP27 in yak reproductive process.