限饲对湖羊子宫RGMb基因及BMP系统成员表达的影响

Effect of Food Restriction on the Expression of RGMb and BMP System Members in the Uterus of Hu Ewes

本研究旨在探讨限饲对湖羊子宫RGMb基因及BMP系统成员表达的影响。首先应用qPCR检测RGMb mRNA在3月龄湖羊机体的组织表达谱,应用IHC检测RGMb在子宫中的定位。然后选取妊娠35d湖羊分为两组,对照组(C)饲喂100%NRC日粮,限饲组(R)饲喂50%NRC日粮,妊娠110天取其子宫组织(n=8),应用qPCR和Western blotting(WB)检测BMP2、BMP4、BMPR1A、BMPR1B、BMPR2与RGMb的表达。结果,RGMb mRNA在湖羊机体的14种组织中均表达,且在子宫的相对表达量大于0.5。湖羊子宫RGMb主要定位于腔上皮与腺上皮细胞。RGMb蛋白在限饲组(R)湖羊子宫中的表达较对照组(C)显著下降(P<0.05),但mRNA水平则无显著变化(P>0.05);限饲组(R)湖羊子宫中的BMP2与BMP4 mRNA水平较对照组(C)显著上升(P<0.05),而BMP受体BMPR1A、BMPR1B与BMPR2mRNA水平较对照组显著下降(P<0.05)。限饲影响子宫组织BMP系统成员尤其是RGMb蛋白的表达,提示RGMb可能参与BMP系统对湖羊子宫稳态的调控,为进一步研究BMP在子宫中的作用机制提供了试验依据。

The study aimed to explore the effects of food restriction on the expression of RGMb and BMP system members in the uterus of Hu ewes. Firstly, qPCR were applied to investigate the RGMb expression profile in different tissues of 3-month-old Hu ewes, IHC were used to detect the expression of RGMb in uterus. Then Hu ewes on 35 days of pregnancy （d35） were divided into 2 groups： ewes in the control （C） group were fed with 100% NRC and ewes in the restriction （R） group were fed with 50% NRC. On 110 days of pregnancy, uterine tissues （n=8） were collected to evaluate the expression of BMP2, BMP4, BMPR1A, BMPR1B, BMPR2 and RGMb by qPCR and WB. qPCR results showed that RGMb mRNA were detected in 14 tissues of 3-month-old Hu ewes, and the relative expression was greater than 0.5 in uterus. IHC staining showed that RGMb protein were mainly expressed in the endometrial luminal and glandular epithelial cells of Hu ewes. The uterine expression of RGMb protein in the R group was significantly lower （P〈0.05） than that in the C group, while there was no significant differences in RGMb mRNA abundance（P〉0.05）. The expression of BMP4 and BMP2 mRNAs in the R group were higher （P〈0.05） than those in the C group. However, the expression of BMP receptors（BMPR1A, BMPR1B and BMPR2） significantly decreased in the R group（P〈0.05）. The results indicate that food restriction affect the expression of BMP system members, including the RGMb, suggesting RGMb accompanied with BMP system may be involved in the regulation of uterine hemeostasis, which will set up experimental evidences for the further study on the regulation mechanism of BMP system in the uterus.