Suh Han Na, Han Ho Jae
Department of Veterinary Physiology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul, South Korea.
Br J Pharmacol. 2015 Feb;172(3):815-28. doi: 10.1111/bph.12947. Epub 2014 Dec 23.
To use stem cell therapy effectively, it is important to enhance the therapeutic potential of stem cells with soluble factors. Although sonic hedgehog (shh) is important in maintaining the stem cell, the recovery effect of mouse embryonic stem cells (mESCs) with shh has not yet been elucidated. The present study investigated the effect of mESCs with shh in skin recovery in vivo as well as the related intracellular signal pathways in vitro.
The healing effect of mESCs with shh on skin wounds was examined in vivo in ICR mice. The involvement of Smads, the microRNA (miR)-200 family, zinc finger E-box-binding homeobox (ZEBs) and E-cadherin on shh-induced mESC migration and self-renewal was determined in vitro.
The mESCs with shh increased re-epithelialization and VEGF expression in skin wounds. Shh-treated mESCs increased both secreted and intracellular levels of VEGF. Shh induced dephosphorylation of glycogen synthase kinase 3β through the Smoothened receptor and increased the phosphorylation of Smad1 and Smad2/3 in mESCs. Shh-induced decrease of the mmu-miR-141, -200c, -200a, -200b and -429 expression levels was significantly reversed by Smad4 siRNA. Shh increased nuclear expression of ZEB1/ZEB2 and decreased E-cadherin expression while increasing cell migration and skin wound healing. Both these effects were reversed by mmu-miR-141 and -200b mimics.
Mouse ESCs accelerated skin wound healing by shh through down-regulating E-cadherin, an effect dependent on mmu-miR-141 and -200b. Our data provides evidence for the effectiveness of shh in stem cell-based therapy in vivo.
为有效利用干细胞疗法，利用可溶性因子增强干细胞的治疗潜力至关重要。尽管音猬因子（shh）在维持干细胞方面很重要，但shh对小鼠胚胎干细胞（mESCs）的恢复作用尚未阐明。本研究调查了mESCs与shh在体内皮肤修复中的作用以及体外相关的细胞内信号通路。
在ICR小鼠体内检测mESCs与shh对皮肤伤口的愈合作用。在体外确定Smads、微小RNA（miR）-200家族、锌指E盒结合同源框蛋白（ZEBs）和E-钙黏蛋白在shh诱导的mESC迁移和自我更新中的作用。
mESCs与shh可促进皮肤伤口的再上皮化和VEGF表达。经shh处理的mESCs增加了VEGF的分泌水平和细胞内水平。shh通过Smoothened受体诱导糖原合酶激酶3β去磷酸化，并增加mESCs中Smad1和Smad2/3的磷酸化。Smad4小干扰RNA可显著逆转shh诱导的mmu-miR-141、-200c、-200a、-200b和-429表达水平的降低。shh增加ZEB1/ZEB2的核表达，降低E-钙黏蛋白表达，同时增加细胞迁移和皮肤伤口愈合。mmu-miR-141和-200b模拟物可逆转这两种作用。
小鼠胚胎干细胞通过shh下调E-钙黏蛋白来加速皮肤伤口愈合，这一作用依赖于mmu-miR-141和-200b。我们的数据为shh在体内基于干细胞的治疗中的有效性提供了证据。
