Slit/Robo通路参与调控小鼠黄体细胞凋亡

Slit/Robo pathway participates in luteal cells apoptosis

本文旨在研究Slit/Robo家族成员在小鼠卵巢组织中的表达及其功能。用real-time PCR检测Slit/Robo家族成员在小鼠卵巢中的mRNA表达丰度,用免疫组织化学方法检测Slit2/Robo1在卵巢组织中的定位,用real-time PCR和免疫组织化学方法检测Slit2/Robo1在不同时期黄体组织中表达的变化,并用Slit/Robo信号通路的阻断剂ROBO1/Fc chimera在体外研究其在小鼠黄体组织中的功能。结果显示,在Slit/Robo家族成员中,配体Slit2和受体Robo1在小鼠卵巢组织中的表达丰度最高,Slit2和Robo1表达定位于小鼠的黄体细胞。与发情前期卵巢相比,间情期卵巢组织中Slit2和Robo1的mRNA表达水平均显著上调(P <0.01, P <0.001)。与妊娠黄体相比,晚期黄体Slit2和Robo1 mRNA表达水平均显著上调。阻断Slit/Robo信号通路后,晚期黄体细胞的凋亡率显著下降(P <0.05)。以上结果提示,Slit/Robo家族成员主要表达于晚期黄体组织,并参与调控黄体细胞的凋亡过程。

This study was aimed to examine the expression and function of Slit/Robo family members in mouse ovary.Real-time PCR was used to assess the mRNA expression levels of Slit/Robo family members,and immunohistochemistry was used to examine the location of Slit2 and Robo1 in the ovary.The mRNA and protein expression levels of Slit2 and Robo1 in early-,middle-and latephase corpus luteum(CL)were examined by real-time PCR and immunohistochemistry,respectively.Blocking agent ROBO1/Fc chimera was used in the luteal cells in vitro to examine the function of Slit/Robo signaling pathway in mouse CL.The results showed that,among the Slit/Robo family members,the expression levels of ligand Slit2 and receptor Robo1 were the highest in mouse ovarian tissue.Moreover,both of them were specifically expressed in mouse luteal cells.Compared with proestrus ovaries,the expression levels of Slit2 and Robo1 mRNA in the ovaries during diestrus were significantly up-regulated(P<0.01,P<0.001).The mRNA expression levels of Slit2 and Robo1 in late-phase CL were significantly increased when compared with pregnant CL.Furthermore,blocking Slit/Robo signaling pathway with ROBO1/Fc chimera in the luteal cells in vitro significantly decreased the apoptotic rate of late luteal cells.These results suggest that Slit/Robo family members are mainly expressed in the late-phase CL of ovary and participate in luteal cells apoptosis.