miR-135a-5p靶向抑制Kif3B调控小鼠腭胚突间充质细胞的自噬

背景：研究表明，在地塞米松诱导腭裂的小鼠胚胎腭突间充质细胞中miR-135a-5p呈高表达，初级纤毛及其介导的Shh信号通路参与小鼠胚胎腭突间充质细胞的自噬。由此猜测miR-135a-5p可能通过初级纤毛及其介导的Shh信号途径调控小鼠胚胎腭突间充质细胞的自噬。目的：探讨miR-135a-5p对小鼠胚胎腭突间充质细胞自噬的调控作用。方法：体外提取并培养C57BL/6J小鼠胚胎腭突间充质细胞。细胞转染分别设置为：(1)对照组、miR-135a-5p阴性对照组、miR-135a-5p模拟物组。(2)NC+miR-NC组、KIF3B过表达组、miR-135a-5p+KIF3B组。qRT-PCR验证miR-135a-5p、KIF3B的转染效率；透射电镜观察各组细胞中自噬小体/自噬溶酶体的数目；免疫荧光技术测定自噬标记物LC3B的荧光表达程度；Western blot检测KIF3B、LC3和P62的蛋白表达。(3)miR-135a-5p阴性对照组、SAG处理组、SAG+miR-135a-5p组。qRT-PCR检测Shh信号下游关键转录因子Gli3的mRNA表达水平；Western blot检测自...     

初级纤毛与Hedgehog信号通路相关性研究

早在20世纪中期以前,初级纤毛被认为是没有任何功能的原始运动纤毛的退化器官.但是近年发现初级纤毛参与了细胞内信号传导等一系列细胞过程,从而调控动物的发育和各种器官的正常生理功能.Hedgehog信号通路对胚胎发育、器官形成具有重要作用.     

Sonic hedgehog信号通路在肺发育及肺癌发生中的意义

在胎肺的发育过程中,支气管上皮细胞和间质之间的通讯联系对胎肺的正常形态构筑以及功能成熟都起到了重要的调控作用。Sonic hedgehog信号通路(Shh信号通路)则在这种联系中发挥着重要作用,其与其他细胞因子相互协调,形成了上皮与间质间调节作用的网络,确保了胎肺的正常发生和发育。近年来,在一些肺癌组织中也发现了Shh信号通路的异常激活,提示Shh信号通路在肺癌的发生发展过程中可能也起到了一定的作用。     

上腭发生的分子调控机制

哺乳动物上腭的发生包括原生腭和次生腭的发生。其中,次生腭的发生涉及到高度动态的形态发生过程,分为腭突生长及模式化,两侧腭突的上抬/重定位,两侧腭突的黏附和融合形成次生腭等过程。近来的研究表明,上腭发育所需的基因,包括了声波刺猬蛋白（Shh）、成纤维细胞生长因子（FGF）、转化生长因子β（TGF-β）和Wnt信号通路,这些信号通路之间存在相互调节,信号通路的异常都会引起腭发育异常,甚至出现腭裂。本文中我们综述了上腭发育过程中各个阶段至关重要的基因及调控网络的研究进展。     

初级纤毛在关节软骨中的生物力学效应

生物力学因素对于关节软骨的稳态维持具有至关重要的作用。初级纤毛是一种可同时感受力学信号和化学信号的细胞器，并于软骨细胞膜表面也存在有初级纤毛分布。其与多个信号转导通路相关，共同参与软骨细胞表型维持和物质代谢的过程。同时，初级纤毛的异常也关联到多种人类的骨关节类疾病。主要论述初级纤毛在软骨细胞力学微环境中的作用，以及与其他信号通路的交互作用机制，探讨其与骨关节疾病的联系，以期为骨科临床和基础科研提供一定的科学依据。     

初级纤毛在肿瘤发生发展中的研究进展

初级纤毛是一种由微管构成的不具有运动功能的细胞器,它参与了细胞的生长发育、内环境稳态、信号转导以及肿瘤的发生和发展。该文就初级纤毛的基本结构、组装与解体,初级纤毛相关的主要信号通路,初级纤毛在肿瘤中的研究进展进行系统性的综述。     

Sonic Hedgehog信号通路在胚胎发育及神经修复中的现状与进展

背景：多项研究表明Sonic Hedgehog (Shh)信号通路可调控神经细胞的增殖、分化和轴突形成,参与脑损伤后的神经再生。 目的：总结Shh信号通路在胚胎发育及出生后神经修复中的作用。 方法：由第一作者检索PubMed 数据库及CNKI全文数据库1980年1月至2015年7月的相关文献,并进行筛选,归纳和总结。英文检索词为“Shh signal pathway, embryogenesis,neural regeneration”,中文检索词为“Shh信号通路,胚胎发育,神经修复”,选择有关Shh在胚胎期细胞分化、组织发育中的研究及出生后参与神经修复,轴突迁移导向及肿瘤发生发展的相关研究,共检索38篇。 结果与结论：近年来Shh信号通路因与脑损伤后神经组织修复的密切关系而备受关注。Shh在Hedgehog (Hh)家族中具有最广泛表达,在胚胎发育、器官形成中起着重要的作用,参与神经系统模式发生、调控前体细胞的分化和迁移、控制轴突的生长和导向,且与肿瘤的发生密切相关。相关研究表明Shh能缩减脑卒中大鼠的脑梗死体积并改善行为学预后。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Sonic Hedgehog及其受体Patched在小鼠视交叉发育过程中的表达

在小鼠胚胎发育过程中,胚胎第13d(E13)至15d(E15)是视交叉发育的主要阶段。在本研究中,我们观察了在E13～E15,Sonic Hedgehog(Shh)及其受体Patched(Ptc)在视觉传导通路的表达。结果发现:在视交叉和视束中,Shh在视神经纤维接近中线时表达上调,越过中线后表达下调,并且主要表达在较深的区域。Ptc在E13～E14的视网膜和E14～E15的视束中有表达,但在视交叉中无表达。Ptc,而不是Shh,表达在体外培养的生长锥中。Shh和Ptc在视觉传导通路发育中的表达提示Shh可能在引导视神经生长方面发挥一定作用。     

白藜芦醇对NIH3T3细胞Shh信号通路的影响

目的探讨白藜芦醇对NIH3T3细胞Shh信号通路的影响。方法实验分为对照组和白藜芦醇组。白藜芦醇处理NIH3T3细胞24 h。CCK-8法测定细胞活力,免疫荧光法检测初级纤毛蛋白(Ac-tu)、Smo和Gli-1的表达,Western blotting法检测Shh、Ptc、Smo、Gli-1蛋白的表达。结果 1.0.5和1μmol/L白藜芦醇组(0.679±0.047,0.774±0.054)细胞活力分别较对照组(0.585±0.039)明显增强(P<0.05),其中以1μmol/L白藜芦醇组最强。之后随白藜芦醇浓度(10、20、40、80μmol/L)的增高,细胞活力逐渐降低(0.428±0.043,0.395±0.031,0.373±0.017,0.361±0.016),且均较对照组明显减弱(P<0.05),而0.1μmol/L(0.602±0.065)和5μmol/L组(0.556±0.041)细胞活力与对照组比较差异无显著性(P>0.05)。2.NIH3T3细胞表达Ac-tu、Shh、Ptc-1、Smo、Gli-1从蛋白,有1根初级纤毛,Smo和Gli-1蛋白位于细胞质。白藜芦醇处理24 h时,Gli-1从细胞质进入细胞核,Smo从细胞质进入初级纤毛,且Shh(0.756±0.659比0.441±0.769,P<0.05)、Ptc-1(0.655±0.347比0.351±0.026,P<0.05)、Smo(0.779±0.064比0.451±0.035,P<0.05)和Gli-1(0.856±0.044比0.560±0.058,P<0.05)蛋白表达较对照组高。结论白藜芦醇可能通过激活Shh信号通路增强NIH3T3细胞的活力。     

Hippo通路在颅面器官发育中的作用

目的介绍Hippo通路在颅面部器官发育相关领域中的最新进展。方法广泛查阅近年来国内外关于Hippo通路各组分在调节颅面器官发育中的作用的相关研究,对相关功能及机制进行总结。结果 Hippo信号通路是由一系列进化保守的蛋白构成激酶级联反应,通过对细胞的增殖,凋亡及分化等多种事件的调节来参与颅面部器官发育。结论脊椎动物颅面部器官的发育需要一系列复杂有序的信号分子共同协调参与。Hippo通路在颅面发育中具有重要作用,有望为探索颅面畸形新的治疗靶点提供帮助。     

BBS proteins interact genetically with the IFT pathway to influence SHH-related phenotypes

There are numerous genes for which loss-of-function mutations do not produce apparent phenotypes even though statistically significant quantitative changes to biological pathways are observed. To evaluate the biological meaning of small effects is challenging. Bardet-Biedl syndrome (BBS) is a heterogeneous autosomal recessive disorder characterized by obesity, retinopathy, polydactyly, renal malformations, learning disabilities and hypogenitalism, as well as secondary phenotypes including diabetes and hypertension. BBS knockout mice recapitulate most human phenotypes including obesity, retinal degeneration and male infertility. However, BBS knockout mice do not develop polydacyly. Here we showed that the loss of BBS genes in mice result in accumulation of Smoothened and Patched 1 in cilia and have a decreased Shh response. Knockout of Bbs7 combined with a hypomorphic Ift88 allele (orpk as a model for Shh dysfuction) results in embryonic lethality with e12.5 embryos having exencephaly, pericardial edema, cleft palate and abnormal limb development, phenotypes not observed in Bbs7(-/-) mice. Our results indicate that BBS genes modulate Shh pathway activity and interact genetically with the intraflagellar transport (IFT) pathway to play a role in mammalian development. This study illustrates an effective approach to appreciate the biological significance of a small effect.     

Commercially available recombinant sonic hedgehog up-regulates Ptc and modulates the cytokine and chemokine expression of human macrophages: an effect mediated by endotoxin contamination?

The Sonic hedgehog (Shh) signalling pathway plays an important role in developmental patterning and proliferation. Recent evidence suggests that Shh also plays a role in the development of the immune system. Here, we demonstrate that components of the Shh signalling pathway are expressed in human macrophages and that the receptor for Shh, Ptc, is up-regulated by a commercially available recombinant preparation of Shh (CArShh). Further, we report that the addition of CArShh up-regulates the production of IL-6, IL-8, MCP-1, IP-10, MIG and RANTES by macrophages, an effect enhanced by the presence of fetal calf serum in the culture medium. In contrast, TGF-beta, TNF-alpha, IL-1b, IL-12 and IL-10 production were not modulated by CArShh and VEGF was minimally up-regulated even in the presence of serum. The up-regulation of these cytokines and chemokines was abrogated by CD14 inhibition and polymixin B, but not reliably inhibited by the specific Shh pathway inhibitor cyclopamine. These results suggest that, although components of the Shh signalling pathway are expressed in macrophages, the modulation of macrophage cytokine and chemokine effector function seen in response to commercially available rShh results from low levels of endotoxin contained within the CArShh preparations employed to explore the effects of Shh in vitro.     

Expression of Sonic hedgehog pathway genes is altered in colonic neoplasia

The Hedgehog (Hh) signalling pathway is crucial for normal development and patterning of numerous human organs including the gut. Hh proteins are also expressed during gastric gland development and gastric epithelial differentiation in adults. Recently, dysregulation of these developmentally important genes has been implicated in cancer, leading to the present study of the expression of Hh signalling proteins in colon cancer. In this study, normal colon and colonic lesions (hyperplastic polyp, adenoma, and colonic adenocarcinoma) were examined by immunohistochemistry using antibodies against Hh signalling molecules: the secreted protein Sonic hedgehog (SHH), its receptor Patched (PTCH), and the PTCH-associated transmembrane protein Smoothened (SMOH). The study shows that Hh signalling pathway members are expressed in normal colonic epithelium. SHH was expressed at the top of the crypts and in a few basally located cells, while PTCH was detected in the neuroendocrine cells and SMOH at the brush border of superficial epithelium. RT-PCR analysis of laser-microdissected crypts from normal human colon confirmed that mRNAs encoding these proteins were expressed in colonic epithelium. Expression of SHH, PTCH, and SMOH was up-regulated in hyperplastic polyps, adenomas, and adenocarcinomas of the colon, and SHH expression correlated with increased expression of the proliferation marker Ki-67 in all lesions examined. To address whether the Hh signalling pathway is functional in the gut, the effect of Shh on epithelial cells in vitro was explored by treating primary murine colonocytes with either Shh peptide or neutralizing anti-Shh antibody. The proportion of cells in the S-phase was assessed by bromodeoxyuridine (BrdU) incorporation. It was found that exogenous Shh promotes cell proliferation in colonocytes, while anti-Shh inhibits proliferation, suggesting that Shh is required during proliferation of epithelial cells in vitro. It is suggested that SHH is required during epithelial proliferation in the colon and that there is a possible role for Hh signalling in epithelial colon tumour progression in vivo.