重组Noggin蛋白对急性肺损伤大鼠肺泡上皮细胞凋亡的影响

目的观察重组Noggin蛋白对酸吸入致急性肺损伤(ALI)大鼠肺泡上皮细胞(AECs)凋亡的影响。方法 30只Wistar雄性大鼠随机均分为对照组(C组)、ALI组(A组)和Noggin组(N组)。麻醉和气管插管后,在吸入相C组给予生理盐水,A组和N组给予盐酸制备模型。然后经尾静脉C组和A组注射生理盐水,N组注射重组Noggin蛋白。造模后6、20 h每组分别处死5只大鼠,取肺组织观察和计数AECs凋亡情况。结果造模后6 h,各组核染色阳性细胞均出现在与肺泡管连接的区域,分布于肺泡壁角落,与Ⅱ型AECs在肺泡内的定位一致;20 h后,许多核染色阳性细胞出现在肺泡腔。定量分析A组造模后6 h和20 h核染色阳性细胞计数明显高于C组,N组核染色阳性细胞数量明显少于A组。结论重组Noggin蛋白明显减轻ALI后AECs凋亡,具有减轻ALI的肺保护作用。     

The function and regulation of TBX22 in avian frontonasal morphogenesis

The frontonasal mass gives rise to the facial midline and fuses with the maxillary prominence to form the upper lip. Here we focus on the regulation and function of TBX22, a repressor dynamically expressed in the frontonasal mass. Both FGF and Noggin (a BMP antagonist) strongly induce gTBX22, however, each has opposite effects on morphogenesis ‐ Noggin inhibits whereas FGF stimulates growth. To determine whether TBX22 mediates these effects, we used retroviruses to locally increase expression levels. RCAS::hTBX22 decreased proliferation, reduced expression of MSX2 and DLX5 and caused cleft lip. Decreased levels of endogenous gTBX22 were also observed but were not the primary cause of the phenotype as determined in rescue experiments. Our data suggest that genetic or environmental insults such as those affecting the BMP pathway could lead to a gain‐of‐function of TBX22 and predispose an individual to cleft lip. Developmental Dynamics 239:458–473, 2010. © 2009 Wiley‐Liss, Inc.     

noggin对帕金森病大鼠黑质神经元的保护作用

目的  观察noggin对6-羟基多巴胺(6-OHDA)致帕金森病模型大鼠神经行为的影响及其对黑质神经元的保护作用。方法  随机将60只雄性SD大鼠,分为正常对照组、模型组和模型治疗组。采用脑立体定位右侧纹状体内注射6-OHDA构建帕金森病(PD)模型,模型治疗组纹状体内注射noggin,正常对照组与模型组同时给予等量生理盐水,术后7d于行为检测完毕后行尼氏染色、TH、TUNEL与GFAP免疫组化染色,光镜下观察3组大鼠黑质多巴胺能神经元、凋亡神经元与星形胶质细胞的形态变化并进行计量分析,电镜下观察黑质神经元超微结构变化。结果    模型治疗组大鼠的旋转次数较模型组明显改善(P<0.05),TH阳性神经元较模型组增多,凋亡神经元显著减少(P<0.05),GFAP阳性神经元较模型组减少,胞浆内GFAP阳性产物的平均光密度值亦明显降低(P<0.05);TH、TUNEL与GFAP阳性神经元主要位于黑质致密部;电镜下对照组黑质神经元结构清晰,胞核形态规则,核膜光滑、完整,染色质分布均匀;胞浆电子密度中等,细胞器丰富;模型组黑质神经元胞核皱缩,核膜凸凹不整、异染色质减少,胞浆内大量线粒体嵴断裂消失,粗面内质网扩张;模型治疗组黑质神经元细胞核形态规则,部分线粒体肿胀,细胞器丰富。结论  noggin可改善PD模型大鼠的行为能力、使凋亡神经元及反应性星形胶质细胞数目减少,这些变化可能与noggin能促进黑质内部分神经元再生有关。     

神经生长因子、Noggin基因转染对骨髓间充质干细胞分化的影响

目的 研究外源性神经生长因子(NGF]、Noggin转染骨髓间充质干细胞(BMSCs)的可行性,并观察基因修饰的细胞向神经元方向的分化情况.方法 采用贴壁法分离纯化BMSCs,通过细胞表面标志及成脂诱导鉴定细胞.Ad-GFP-NGF、Ad-GFP-Noggin单独及联合转染BMSCs.通过Westernblot、免疫细胞化学观察目的 蛋白表达.通过免疫组化观察转染后BMSCs向神经元方向分化情况.结果贴壁法获得的细胞具有BMSCs表型,能分化为脂肪细胞.未转染组和Ad-GFP转染组少量表达NGF,不表达Noggin.NGF、Noggin单独及联合转染BMSCs均能高效表达目的 蛋白.NGF、Noggin转染的BMSCs可分化为具有神经元形态,并表达神经丝蛋白(NF-H)的细胞,联合转染组NF-H阳性细胞比例最高.结论 贴壁法能有效纯化BMSCs,Ad-GFP-NGF、Ad-GFP-Noggin单独及联合转染BMSCs安全并能高效表达目的 蛋白,NGF、Noggin转染的BMSCs 体外培养能向神经元样细胞方向分化,两种蛋白联合修饰能增强这种分化作用.     

Noggin在大鼠中枢神经系统发育过程中的表达

目的 研究Noggin基因在大鼠中枢神经系统(CNS)发育过程中的表达。方法 地高辛标记的cRNA探针原位杂交组织化学技术。结果 ISHH结果显示，在胚胎期(E16)大鼠，noggin mRNA阳性细胞主要位于大脑皮质、海马、丘脑与下丘脑的部分核团。新生期(P1-P2)大鼠，noggin在大脑皮质与海马的表达均降低，而在丘脑与延脑的表达增强；生后1周(P1W)noggin在脑内的表达明显降低，生后2周noggin在脑内的表达开始升高，在大脑皮质与海马升高尤为明显。生后1个月，noggin表达继续升高，在额叶皮质、顶叶皮质、扣带皮质、梨状皮质及海马的齿状回可检测到强阳性信号；而在丘脑的侧核、网状核、腹内侧核与腹外侧核可见中等强度的阳性信号。此外，在下丘脑的室旁核和视上核亦可见密集深染的阳性神经元。生后3个月，noggin阳性细胞在脑内的表达开始降低；生后18个月，noggin表达降至最低，仅见散在的阳性神经元。此外，在不同发育期大鼠的脊髓亦未观察到noggin mRNA阳性细胞。结论 提示noggin基因参与大鼠生后CNS的发育。     

Chordin and noggin expression in the adult rat trigeminal nuclei

Bone morphogenetic proteins (BMP) exert its biological functions by interacting with membrane bound receptors. However, functions of BMPs are also regulated in the extracellular space by secreted antagonistic regulators, such as chordin and noggin. Although the deep involvement of BMP signaling in the development and functions of the trigeminal nuclei has been postulated, little information is available for its expression in the trigeminal nuclei. We, thus, investigated chordin and noggin expression in the adult rat trigeminal nuclei using immunohistochemistry. Chordin and noggin were intensely expressed throughout the trigeminal nuclei. In addition, interesting differences are observed between chordin expression and noggin expression. For example, chordin prefers dendritic expression than noggin, suggesting that chordin is involved in the regulation of dendritic morphology and synaptic homeostasis. Furthermore, chordin and noggin were differentially expressed in the neuropil of the trigeminal nuclei. Since BMP signaling is known to play a pivotal role to make precise neural network, theses differences might be important to keep precise interneuronal connections by regulating local BMP signaling intensity in each region. Interestingly, we also detected chordin and noggin expression in axons of the trigeminal nerves. These data indicate that chordin and noggin play pivotal roles also in the adult trigeminal system.     

BMP-4 and Noggin signaling modulate dorsal fin and somite development in the axolotl trunk.

BMP-4, a member of the TGF-beta superfamily of growth factors, is involved in various developmental processes. We investigated the effects of BMP-4 and its antagonist Noggin on axolotl trunk development. Implantation of BMP-4-coated microbeads caused inhibition of muscle and dorsal fin formation in the vicinity of the microbeads. At some distance, myotomes developed with reduced height but increased width, which was accompanied by increased cell proliferation. These effects could be modulated by co-implanting Noggin-coated beads. Immunostaining of Pax7 further revealed that although the dermomyotome was absent in the vicinity of BMP-4-coated beads, at some distance from them, it was thicker than in controls, indicating that moderate amounts of BMP-4 stimulate this layer of undifferentiated cells. In contrast, Noggin generally inhibited the dermomyotome, possibly indicating premature differentiation of dermomyotome cells. We conclude that BMP-4 and Noggin are involved in the regulation of cell proliferation and differentiation during somite development.     

Noggin诱导泌乳素腺瘤细胞凋亡的实验研究

目的:探讨骨形态生成蛋白4(BMP-4)拮抗剂Noggin诱导人泌乳素(PRL)腺瘤细胞的凋亡作用.方法:用MTT法和流式细胞仪检测研究Noggin诱导细胞凋亡的情况;用免疫组化和RT-PCR检测分析Noggin诱导细胞凋亡后对BMP-4蛋白和BMP-4 mRNA的影响.结果:MTT法测定Noggin对PRL腺瘤细胞的IC50值为11μg/mL,不同浓度的Noggin对PRL腺瘤细胞具有抑制作用,并呈现剂量-效应关系.Noggin处理PRL腺瘤细胞后,AnnexinV标记的凋亡细胞增多,电镜观察细胞凋亡增多,同时Noggin可使BMP-4 mRNA及BMP-4蛋白表达下降.结论:Noggin能诱导PRL腺瘤细胞发生凋亡,可有抗PRL腺瘤的作用.     

BMP signaling regulates the fate of chondro‐osteoprogenitor cells in facial mesenchyme in a stage‐specific manner

Background : Lineage tracing has shown that most of the facial skeleton is derived from cranial neural crest cells. However, the local signals that influence postmigratory, neural crest‐derived mesenchyme also play a major role in patterning the skeleton. Here, we study the role of BMP signaling in regulating the fate of chondro‐osteoprogenitor cells in the face. Results : A single Noggin‐soaked bead inserted into stage 15 chicken embryos induced an ectopic cartilage resembling the interorbital septum within the palate and other midline structures. In contrast, the same treatment in stage 20 embryos caused a loss of bones. The molecular basis for the stage‐specific response to Noggin lay in the simultaneous up‐regulation of SOX9 and downregulation of RUNX2 in the maxillary mesenchyme, increased cell adhesiveness as shown by N‐cadherin induction around the beads and increased RA pathway gene expression. None of these changes were observed in stage 20 embryos. Conclusions : These experiments demonstrate how slight changes in expression of growth factors such as BMPs could lead to gain or loss of cartilage in the upper jaw during vertebrate evolution. In addition, BMPs have at least two roles: one in patterning the skull and another in regulating the skeletogenic fates of neural crest‐derived mesenchyme. Developmental Dynamics 245:947–962, 2016. © 2016 Wiley Periodicals, Inc.