Rho/ROCK信号通路与糖尿病肾病

糖尿病肾病(DN)是糖尿病最严重的微血管并发症之一,并已成为终末期肾脏病(ESRD)的重要原因,成为人类致死、致残的一个重要因素。因此,发现糖尿病肾病的新机制及关于此机制的新药研究,对改善糖尿病肾病预后非常重要。近年来,不断深入的研究提示,Rho/ROCK信号通路可能成为防治糖尿病肾病的药物新靶点。本文就Rho/ROCK信号通路与糖尿病肾病的关系作一综述。     

SHH信号通路在海马神经可塑性及相关神经系统疾病中的作用

音猬因子（sonic hedgehog,SHH）是一种分泌蛋白质,可在发育过程中控制神经祖细胞、神经元和神经胶质细胞的形成。研究发现,海马是学习和记忆中至关重要的大脑区域,SHH在海马神经元回路的形成和可塑性中发挥重要作用,可介导海马神经的发生和突触的可塑性调节。海马神经元树突中SHH受体的激活是跨神经元信号通路的组成部分,该信号通路可加速轴突的生长并增强谷氨酸从突触前末端的释放。SHH信号通路转导受损可导致中枢神经系统损伤和相关疾病（如自闭症、抑郁症和神经退行性疾病等）发生。因此,控制SHH信号通路转导,如使用SHH通路抑制剂或激动剂可能有助于相关疾病的治疗。综述了SHH信号通路的海马神经可塑性及其在中枢神经系统发育和相关疾病中的影响,以期为阐明SHH信号转导受损导致的海马神经受损和中枢神经系统相关疾病的机制奠定一定的理论依据。     

RhoA/ROCK信号通路对血管平滑肌收缩的调节作用及研究进展

血管平滑肌的异常收缩是引起许多疾病的重要因素,如高血压,脑血管痉挛等,对于平滑肌收缩调节机制的研究为治疗这些疾病带来新的思路和方向.研究表明小GTP结合蛋白RhoA及其下游信号分子ROCK在平滑肌收缩调节,尤其是钙敏化调节机制中起到关键作用.RhoA/ROCK通路通过抑制MLCP活性而增强MLC的磷酸化水平,从而调节平滑肌收缩,此外,它还参与调节其它细胞的多种细胞功能,如应力纤维的生成,细胞分裂及迁移等.本综述主要介绍RhoA/ROCK通路在血管平滑肌收缩功能的调节机制及研究进展.     

突触可塑性与脑疾病的神经发育基础

大脑神经回路高度有序的神经元活动是高级脑功能的基础,神经元之间的突触联结是神经回路的关键功能节点。神经突触根据神经元活动调整其传递效能的能力,亦即突触可塑性,被认为是神经回路发育和学习与记忆功能的基础。其异常则可能导致如抑郁症和阿尔茨海默病等精神、神经疾病。将介绍这两种疾病与突触可塑性的关系,聚焦于相关分子和细胞机制以及新的研究、治疗手段等进展。     

Rho/Rho激酶信号通路与脉管收缩

Rho/Rho激酶信号通路在正常血管、淋巴管等脉管收缩过程中发挥重要的调节作用,并参与休克后血管反应性和钙敏感性的双相调节以及休克淋巴管低反应性的发生.以Rho/Rho激酶为靶点,对于干预休克脉管系统低反应性的发生具有重要意义.本文综述Rho/Rho激酶信号通路在脉管收缩中的调节机制.     

鸟类鸣啭学习神经回路的发育可塑性

鸟类鸣啭控制系统已成为人们研究神经系统与学习、行为和发育关系的重要模型。鸣啭系统在发育中所表现出的神经和行为明显变化的特点,为我们理解语言学习敏感期、突触联系的再分布、结构特化以及细胞死亡与神经发生提供了宝贵的信息。在许多方面,鸣禽鸟啭系统都有别于哺乳动物,这为特定理论问题的研究提供了新的途径。     

与突触可塑性相关的神经振荡和信息流研究

作为一种有节律的神经活动,神经振荡现象发生在所有的神经系统中,例如大脑皮层、海马、皮层下神经核团以及感觉器官.本综述首先给出了已有的研究结果,即基于theta和gamma频段的同步神经振荡揭示了认知过程的起源与本质,如学习与记忆.然后介绍了关于神经振荡分析的新技术和算法,如表征神经元突触可塑性的神经信息流方向指数,并例...     

大豆异黄酮基于RhoA/ROCK2信号通路改善MCAO大鼠神经功能损伤

目的:探讨大豆异黄酮对脑缺血再灌注大鼠RhoA/ROCK2信号通路介导的氧化应激反应和神经元凋亡的影响。方法:60只SD大鼠随机分为3组,对照组、模型组、大豆异黄酮组。连续给药7天后,给药剂量200 mg/kg。应用中动脉栓塞再灌注模型致大鼠缺血损伤。24 h后评价大鼠神经功能,TTC染色检测脑梗死体积,试剂盒检测脑中氧化因子含量,免疫组化检测神经元损伤,Western Blotting检测RhoA/ROCK2相关蛋白含量。结果:与对照组比较,模型组大鼠神经功能评分降低(P0.05),脑梗死体积增加(P0.05),氧化因子含量增加(P0.05),神经元凋亡显著(P0.05),RhoA/ROCK2蛋白表达增加(P0.05)。与模型组相比,大豆异黄酮升高了大鼠神经功能评分(P0.05),减少的脑梗死体积(P0.05),降低脑中氧化因子含量(P0.05),抑制了神经元凋亡(P0.05),抑制了RhoA/ROCK2蛋白表达(P0.05)。结论:大豆异黄酮可以缓解脑缺血再灌注损伤介导的氧化应激及细胞凋亡,进而减轻神经功能障碍,其机制可能与抑制RhoA/ROCK2信号通路相关。     

鸣禽前端脑通路与鸣唱可塑性

前端脑通路即鸣禽的基底神经节——前脑通路,为鸣唱学习和可塑性所必需。本文综述了前端脑通路的起源、发育、作用及其鸣唱可塑性方面的最新进展。     

OPN通过VEGF途径增强脑卒中后的神经可塑性

该研究旨在探讨骨桥蛋白(osteopontin,OPN)对缺血性脑卒中亚急性期的神经可塑性作用。制备小鼠光化学栓塞模型,在卒中后2、7、14、21、28天采用Western blot测定小鼠大脑皮层梗死周区组织中突触素(synaptophysin,SYN)的蛋白表达水平。卒中后第7天,通过脑立体定位仪局部给予人重组骨桥蛋白(recombinant osteopontin,rOPN)。运用囊泡谷氨酸转运体1(vesicular glutamate transporter 1,VgluT 1)、突触后致密蛋白95(post-synaptic density protein 95,PSD 95)免疫荧光染色,观察骨桥蛋白治疗后梗死周区皮层共存突触的数量。采用胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、离子化钙结合适配器分子1(ionized calcium-binding adapter molecule 1,IBA1)、血小板内皮黏附分子31(platelet endothelial cell adhesion molecule-1,CD3...     

Roles for Rho/ROCK and Vinculin in Parietal Endoderm Migration

The first cell migration event in the mouse embryo is the movement of parietal endoderm cells from the surface of the inner cell mass facing the blastocoel cavity to line the inner surface of the trophectoderm. F9 embryoid bodies provide an in vitro model for this event. They have an inner core of undifferentiated stem cells surrounded by an outer visceral endoderm layer. When plated on a laminin coated substrate, visceral endoderm transitions to parietal endoderm and migrates onto the dish, away from the attached embryoid body. We now show that this outgrowth contains abundant focal complexes and focal adhesions, as well as lamellipodia and filopodia. Treatment with the ROCK inhibitor Y-27632 promotes a 2-fold increase in outgrowth, and a transition from focal adhesions and associated stress fibers, to focal complexes and a decrease in stress fibers. ROCK inhibition also leads to an increase in lamellipodia. Inhibition of RhoA by transfection of a vector encoding C3 transferase, direct administration of the C3 enzyme, or transfection of a vector encoding p190 Rho GTPase Activating Protein also promotes outgrowth and an apparent transition from focal adhesions to focal complexes. Parietal endoderm outgrowth generated using vinculin-deficient F9 stem cells migrates 2-fold further than wild type cultures, but this outgrowth retains the morphology of wild type parietal endoderm, including focal adhesions and stress fibers. Addition of Y-27632 to vinculin-null outgrowth cultures further stimulates migration an additional 2-fold, supporting the conclusion that Rho/ROCK and vinculin regulate parietal endoderm outgrowth by distinct pathways.     

Discovery and optimization of indole and 7-azaindoles as Rho kinase (ROCK) inhibitors (part-II)

Therapeutic interventions with Rho kinase (ROCK) inhibitors may effectively treat several disorders such as hypertension, stroke, cancer, and glaucoma. Herein we disclose the optimization and biological evaluation of potent novel ROCK inhibitors based on substituted indole and 7-azaindole core scaffolds. Substitutions on the indole C3 position and on the indole NH and/or amide NH positions all yielded potent and selective ROCK inhibitors (25, 42, and 50). Improvement of aqueous solubility and tailoring of in vitro and in vivo DMPK properties could be achieved through these substitutions.     

Rho激酶抑制剂在神经退化性疾病上的应用

Rho激酶,又称Rho相关的卷曲蛋白激酶,是一类丝氨酸/苏氨酸蛋白激酶,被发现为小G蛋白Rho的下游作用底物。由于Rho激酶活性涉及神经细胞的功能,而且越来越多的研究表明抑制Rho激酶的活性在数种神经退行性疾病包括帕金森病、阿尔茨海默病、亨廷顿病、多发性硬化症,和肌萎缩性侧索硬化症等的实验模式中都有明显的效果。因此,Rho激酶已成为针对治疗神经性退化性疾病的一个热门标靶蛋白。本文探讨Rho激酶抑制剂在神经退化性疾病上的应用及发展,使神经退行性疾病能进一步提升治疗和在应用上的水平。     

Discovery and optimization of indoles and 7-azaindoles as Rho kinase (ROCK) inhibitors (part-I)

Rho kinase (ROCK) inhibitors are potential therapeutic agents to treat disorders such as hypertension, multiple sclerosis, cancers, and glaucoma. Here, we disclose the synthesis, optimization, biological evaluation of potent indole and 7-azaindole based ROCK inhibitors that have high potency on ROCK (IC(50)=1 nM) with 740-fold selectivity over PKA (47). Moreover, 47 showed very good DMPK properties making it a good candidate for further development. Finally, docking studies with a homology model of ROCK-II were performed to rationalize the binding mode of these compounds and showed the compounds bound in both orientations to take advantage to H-bonds with Lys-121 of ROCK-II.     

Alkannin attenuates lipopolysaccharide‐induced lung injury in mice via Rho/ROCK/NF‐κB pathway

We investigated the effects and associated mechanism of alkannin (AL) on lipopolysaccharide (LPS)‐induced acute lung injury in a mouse model. Pretreatment with AL in vivo significantly reduced the lung wet/dry weight ratio and inhibited lung myeloperoxidase activity and malondialdehyde content, while increasing superoxide dismutase activity. Hematoxylin and eosin staining demonstrated that AL attenuated lung histopathological changes. In addition, AL‐inhibited overproduction of proinflammatory cytokines in bronchoalveolar lavage fluid and lung tissues in LPS‐injured mice and LPS‐exposed A549 cells. Further analysis showed that AL‐inhibited induction of the Rho/ROCK/NF‐κB pathway via LPS‐induced inflammation in mice and A549 cells. Fasudil, a selective ROCK inhibitor, showed similar effects. Overall, the findings indicate that AL suppresses the expression of messenger RNAs and proteins associated with Rho/ROCK/NF‐κB signaling to effectively ameliorate lung injury.     

Synthesis and biological evaluation of 4-quinazolinones as Rho kinase inhibitors

Rho kinase (ROCK) is an attractive therapeutic target for various diseases including glaucoma, hypertension, and spinal cord injury. Herein, we report the development of a series of ROCK-II inhibitors based on 4-quinazolinone and quinazoline scaffolds. SAR studies at three positions of the quinazoline core led to the identification of analogs with high potency against ROCK-II and good selectivity over protein kinase A (PKA).     

Possible participation of the Rho/Rho-associated coiled-coil-forming kinase pathway in the cell death of Cryptococcus neoformans caused by Staphylococcus aureus adherence

We here report the apoptotic death of a fungus, Cryptococcus neoformans (C. neoformans), in response to adherence of the pathogenic bacterium Staphylococcus aureus (S. aureus). In co-culture, cryptococcal actin was visibly aggregated. To investigate the mechanism of death, the participation of small GTP(guanosine triphosphate)-binding proteins belonging to the Rho subfamily, which regulate the actin cytoskeleton, was explored. C. neoformans was cultured with S. aureus in the presence of N-(4-pyridyl)-4-(1-aminoethyl)cyclohexanecarboxamide (Y-27632), an inhibitor of Rho-associated coiled-coil forming kinase (ROCK), a downstream effector of Rho. Death of C. neoformans was significantly reduced by the inhibitor. Concomitantly, Y-27632 prevented the aggregation of actin. Therefore, it was concluded that the Rho/ROCK pathway is involved in cell death induced by adherence stress. Increased expression of the voltage-dependent anion channel (VDAC), located in the mitochondrial outer membrane, has previously been observed in the apoptosis-like death of C. neoformans in the presence of hydrogen peroxide. Ruthenium red (RuR), which binds to VDAC and inhibits cytochrome c release, was used to determine the involvement of VDAC following adherence stress caused by S. aureus. RuR treatment increased the viability of C. neoformans co-cultured with S. aureus in a dose dependent manner. These findings suggest that Rho-ROCK signaling could be involved, via a mitochondrial pathway, in the apoptosis-like death of C. neoformans induced by the adherence of S. aureus.     

ROCK inhibitors 3: Design,synthesis and structure-activity relationships of 7-azaindole-based Rho kinase (ROCK) inhibitors

Rho kinase (ROCK) inhibitors are potential therapeutic agents for the treatment of a variety of disorders including hypertension, glaucoma and erectile dysfunction. Here we disclose a series of potent and selective ROCK inhibitors based on a substituted 7-azaindole scaffold. Substitution of the 3-position of 7-azaindole led to compounds such as 37, which possess excellent ROCK inhibitory potency and high selectivity against the closely related kinase PKA.