Rho/Rho激酶信号通路与急性肺损伤/急性呼吸窘迫综合征

Rho/Rho激酶信号通路参与调节细胞的收缩、黏附、迁移、增殖、凋亡等多种生物学行为和功能,随着对Rho/Rho激酶信号通路研究的深入,发现其在急性肺损伤/急性呼吸窘迫综合征中发挥着重要的作用。笔者就Rho/Rho激酶信号通路在急性肺损伤/急性呼吸窘迫综合征中的作用机制作一综述。     

Rho/Rho激酶信号通路异常激活与心血管疾病

Rho蛋白是一种小分子三磷酸鸟苷结合蛋白,目前已从哺乳动物中分离出20种Rho蛋白家族成员。骨骼肌细胞肌动蛋白组合和细胞运动主要受Rho激酶调节。Rho/Rho激酶信号通路是在体内普遍存在,它通过调节细胞内肌动蛋白骨架的聚合状态参与多种细胞功能,包括细胞收缩、迁移、黏附、增殖、凋亡及基因表达等。Rho/Rho激酶信号通路的异常启动在心血管疾病的发病机制和病理生理中发挥了重要作用,对此信号转导通路的研究可为心血管的预防和治疗提供新的靶点。     

Rho/Rho激酶信号通路在妊娠过程中的作用

Rho通过三磷酸鸟苷(GTP)结合形式和二磷酸鸟苷(GDP)结合形式的转换起着分子开关的作用,调控许多细胞内信号通路。Rho通过激活其下游靶分子Rho激酶调节细胞的黏附、迁移、增殖、收缩等多种生物学行为和功能。近年来,越来越多的研究发现Rho/Rho激酶通路与胚胎种植、妊娠维持及分娩发动密切相关,本文对其在妊娠过程中的作用进行综述。     

Rho／Rho激酶信号转导通路与心力衰竭的研究进展

Rho／Rho激酶信号通路的关键信号分子包括Rho蛋白、Rho激酶和肌球蛋白磷酸酶。Rho蛋白是Ras超家族中小分子G蛋白的成员之一,具有GTP酶活性。Rho．GTP在细胞的信号转导通路中作为信号转换器或分子开关,与Rho激酶的Rho蛋白结合域结合,     

Rho抑制剂在冠心病介入治疗中临床应用

自从1985年首次发现rho(ras homologous)基因以来,对其基因产物Rho及其信号传导通路Rho/Rho激酶(Rhokinase,ROK或Rho-associated coiled-coil-containing proteinkinase,ROCK)的病理生理学已进行了广泛的研究。Rho/ROCK信号通路的主要生理功能为调节细胞骨架蛋白的合成、降解、移动和收缩等,并参与细胞的基本生命过程,如迁移、增殖和存活。Rho激酶为Rho蛋白下游作用底物,     

Rho A/Rho激酶信号通路参与慢性应激大鼠抑郁模型结肠动力的改变

目的:检测Rho A、Rock1、Rock2在抑郁大鼠结肠组织中mRNA及蛋白的表达,探讨Rho A/Rho激酶信号通路在抑郁大鼠结肠动力障碍中的可能机制.方法:通过慢性应激建立大鼠抑郁模型,正常组作对照,应用Real Time Q-PCR和Western blot检测Rho A、Rock1、Rock2在结肠组织中mRNA及蛋白的表达.结果:抑郁组结肠组织中的Rho A、Rock1、Rock2的mRNA及蛋白表达明显降低,与正常组比较差异有统计学意义(P＜0.01).结论:RhoA/Rho激酶信号通路在抑郁大鼠结肠动力障碍起着重要作用,但其作用机制仍需进一步的实验研究.     

Rho/Rho激酶与心血管疾病

Rho蛋白是一种三磷酸鸟苷(GTP)结合蛋白,Rho激酶(ROCK)是最早发现的Rho效应物,在分子水平,ROCK表达上调促进氧化应激、炎症、血栓形成和纤维化的各种因子增加,下调内皮型一氧化氮合酶。在细胞水平,参与许多细胞功能如基因表达、胞质分裂、细胞黏附和迁移等的调节。研究发现,Rho/Rho激酶与心血管疾病有关,如冠状动脉粥样硬化性心脏病、高血压、心力衰竭等,其抑制剂fasudil可以治疗许多心血管疾病,目前已经在临床上应用。该文对Rho/Rho激酶与心血管疾病关系予以综述。     

COPD相关性肺动脉高压与Rho/Rho激酶信号通路

慢性阻塞性肺疾病（COPD）是一种常见病、多发病,表现为不完全可逆的气流受限。肺动脉高压（PAH）是一组少见的、预后不良的疾病,以增高的肺动脉压力和阻力为特征。慢性缺氧及慢性炎症是COPD相关性PAH的主要发病原因。目前已证实Rho/Rho激酶通路参与PAH的病理生理过程,在PAH的动物模型中阻断该信号通路能够改善疾病的病理改变与预后。     

低氧性肺动脉高压大鼠肺组织Rho激酶的表达及意义

目的 探讨低氧性肺动脉高压(HPH)大鼠肺组织小G蛋白Rho相关激酶(Rho激酶)的表达及其抑制剂法舒地尔对HPH的影响. 方法 24只雄性SD大鼠随机均分为对照组,低氧模型组和法舒地尔干预组.测各组大鼠平均肺动脉压力(mPAP)、右心室肥厚指数(RVHI);采用逆转录酶-聚合酶链反应(RT-PCR)、免疫印迹法(Western blot),从基因、蛋白质水平观察肺组织Rho激酶表达变化;免疫印迹法检测其底物肌球蛋白磷酸酶的磷酸化状态,作为该激酶功能活化的标志. 结果 低氧模型组Rho激酶mRNA和蛋白明显升高,肌球蛋白磷酸酶磷酸化水平也较对照组显著增高,并与mPAP及RVHI均呈明显正相关(均P＜0.01).法舒地尔干预后mPAP、RVHI、Rho激酶mRNA和蛋白、肌球蛋白磷酸酶磷酸化水平均明显降低.结论 Rho/Rho激酶信号通路在HPH发病中具有重要作用,法舒地尔通过抑制Rho激酶的表达,对低氧所致的肺动脉高压、右心室肥厚具有较好的预防和逆转作用.     

Rho激酶的抑制可改善冠状动脉疾病患者的内皮功能

基础生物学研究提示Rho／Rho激酶通路激活可降低一氧化氮（NO）的生物利用度，进而促发动脉粥样硬化及其临床并发症。然而，关于动脉粥样硬化患者中Rho／Rho激酶通路与N0生物利用度之间的关系甚少报道。因此，作者拟确定Rho激酶的抑制是否可增加冠状动脉疾病（CAD）患者的NO生物利用度，改善其内皮功能。在一项双盲交叉对照试验中，13例CAD患者和16例年龄、性别与之匹配的健康对照者被随机入选接受Rho激酶抑制剂法舒地尔或安慰剂治疗各1月。     

Rho/ROCK2通路在阿尔茨海默病病理进程中的重要作用

Rho/Rho相关卷曲螺旋形成蛋白激酶（ROCK）通路是生物体中广泛存在的经典信号通路,参与多种生理调节过程。ROCK有2种亚型,即ROCK1和ROCK2。在阿尔茨海默病（AD）患者脑中,Rho/ROCK2表现为活性上调,并伴有Aβ42水平升高,以及神经细胞突起的形态与功能异常,推测AD的发生、发展与Rho或ROCK2的高表达或过度激活有关。目前Rho/ROCK2通路被认为是预防和治疗AD的一个有效的靶通路,而Rho或ROCK2,也成为药物研发的重要靶点。研究发现,降低Rho或ROCK2的表达,或者抑制Rho或ROCK2的活性均可减少Aβ42诱导的神经毒性,保护神经元,减缓AD的发展。因此,Rho/ROCK2的特异性抑制对中枢神经损伤修复及AD的治疗有重要的意义。为此,本文针对Rho/ROCK2通路在阿尔茨海默病发展中的作用做一综述。     

中枢神经系统炎性脱髓鞘专病临床科研数据库设计与构建

目的/意义 构建中枢神经系统(central nervous system,CNS)炎性脱髓鞘专病数据库,为临床科研服务,提高基层医疗水平。方法/过程 利用互联网收集病历数据,经过处理和分析后,构建CNS炎性脱髓鞘专病数据库。运用统计分析、自然语言处理、人工智能影像识别和数据可视化等技术,整合分析数据库信息。结果/结论 形成标准化的CNS炎性脱髓鞘专病数据库大数据信息池,将临床科研数据可视化,兼顾患者宣教、专科医生培训、多中心远程会诊功能,促进医疗科研成果转化,为未来真实世界的临床研究提供参考依据,优化诊疗路径,赋能基层医院。     

Smooth muscle contraction by small GTPase Rho

Abnormal contraction of vascular smooth muscle contributes to a variety of diseases such as hypertension and vasospasm in coronary and cerebral arteries. An increment in a cytoplasmic Ca2+ concentration is the key event in smooth muscle contraction. However, smooth muscle contraction is modified upon the stimulation by agonists as well as in some pathophysiological situations in Ca(2+)-independent mechanism. The molecular mechanism underlying this modulation was not elucidated. Recent studies have shown the important role of small GTPase Rho and its effector, Rho-associated kinase (Rho-kinase)/ROK/ROCK in Ca(2+)-independent regulation of smooth muscle contraction. The Rho/Rho-kinase pathway modulates the phosphorylation level of myosin light chain (MLC) of myosin II, mainly through the inhibition of myosin phosphatase, and contributes to the agonist-induced Ca(2+)-sensitization in smooth muscle contraction. The Rho/Rho-kinase pathway is involved in the pathogenesis of hypertension, vasospasm and arteriosclerosis, and is a potent target of new therapies for these diseases.     

Rho/ROCK通路与NADPH氧化酶在糖尿病肾病中的作用

Rho/ROCK通路与NADPH氧化酶的激活,都能够促进糖尿病肾病的发生发展。Rho/ROCK通路与NADPH氧化酶可能也存在着某些联系,研究二者之间的关系与相关机制,对发现治疗糖尿病肾病的新药可能具有重要的意义。本文首先介绍了Rho/ROCK通路与NADPH氧化酶及二者的激活因素,然后总结了二者在糖尿病肾病中对肾脏血流动力学、滤过功能以及肾内基质的影响,最后简要介绍了与二者相关的糖尿病肾病药物治疗靶点。     

Effect of Fasudil on Remyelination Following the Cuprizone-Induced Demyelination in Male C57BL/6 Mice

Background Multiple sclerosis (MS) is an autoimmune, inflammatory demyelinating disease of the central nervous system (CNS) characterized by de-/ remyelination, neuroinflammation and oligodendrocyte loss. Although a greater understanding of MS have increased acquaintance of the pathogenesis and pathophysiology, the exploration of treatment is still challenging. Fasudil, one of the most thoroughly studied Rho kinase (ROCK) inhibitors, has been shown to have effects in neurodegenerative diseases. However, the effect of Fasudil on preventing the progression of the demyelination in MS has not been evaluated. Cuprizone (CPZ)-induced demyelination is a model used to study de-/ remyelination in the CNS. Some aspects of the histological pattern induced by CPZ are similar to MS. The aim of the study is to investigate the effect of Fasudil on CPZ-induced demyelination, and to explore the mechanisms for the possible remyelination. Materials and Methods Male C57BL/6 mice (10–12 weeks old) were assigned into normal group, fed a normal diet; CPZ group, fed CPZ and intraperitoneally (i.p.) injected with normal saline after 4 weeks for consecutive 2 weeks; Fasudil-treated CPZ group, which were i.p. injected with Fasudil (40 mg/kg/day) after 4 weeks for consecutive 2 weeks. All groups were assessed by Elevated plus-maze (EPM) test and Pole test at the end of the experiment. For examing the extent of demyelination, Luxol Fast Blue (LFB) staining, Black Gold II and myelin basic protein (MBP) immunohistochemistry staining were used for slides of brains. Splenic MNCs were fixed and stained with the following antibodies: Alexa Fluor B220, FITC-CD4/PE-IFN-γ, FITC-CD4/PE-IL-17. At least 10, 000 events were collected using flow cytometer. Results Following CPZ-exposure, mice presented a lower density of LFB, Black Gold II and MBP expression, loss of mature oligodendrocytes. Spleen atrophy was observed in CPZ-group compared to normal mice, and we firstly found that CPZ feeding induced the formation of MOG antibody. Fasudil treatment improved behavioral abnormality, promoted remyelination, inhibited spleen atrophy and production of MOG antibodies, prevented the infiltration of peripheral T cells, B cells, macrophages, and declined the neuroinflammation by inhibiting Iba1+iNOS+, Iba1+NF-κB+ microglia. Fasudil treatment also reduced the levels of IL-1β, IL-6 and TNF-α. Discussion In this study, we demonstrated that demyelinating model was successfully established. Then we tested whether Fasudil plays a remyelinating role in this model. Spleen atrophy was observed after CPZ-feeding compared to normal mice. Previous studies have shown that splenic atrophy in experimental stroke may contribute to brain injury possibly through the release of inflammatory mediators and spleen-derived inflammatory cells to the circulation and migration into the brain, which aggravate the brain inflammatory response and led to secondary injure. At present, we lack direct evidence to elucidate the mechanisms for spleen atrophy in CPZ-induced demyelination. We firstly found that CPZ-feeding induced the formation of MOG antibody. Recent study indicated that BBB hyperpermeability precedes demyelination in CPZ-demyelinating model. Another study suggested that debris of damaged cells in the CNS may present as antigens after penetrating the BBB, giving rise to autoantibodies. Therefore, it is possible that the myelin debris produced the destruction of myelin sheath can enter the blood circulation and stimulate the immune response of T and B cells. We found that MOG antibody was elevated in the supernatant of cultured plenocytes, indicating that the MOG antibodies were derived from peripheral immune cells. Our results showed that the level of MOG antibody in the brain homogenate of CPZ-treated mice was higher than that of normal mice, suggesting that antibodies can enter brain tissue and anti a-synuclein antibody was negative, which indicate that anti MOG antibody is a specific antibody. In our study, MOG antibody was capable of being detected in the brain of CPZ-treated mice, providing a possibility for specific MOG antibody-mediated oligodendrocyte damage. CPZ induced a wide range of Iba-1+ microglia, which was inhibited by Fasudil. These results suggest that the suppression of inflammatory microenvironment may contribute to the remyelination. In conclusion, the administration of Fasudil promoted remyelination by multiple mechanisms.     

RhoA/ROCK信号通路对TLR-2配体诱导的类风湿关节炎患者成纤维滑膜细胞分泌趋化因子的调控作用

目的 研究RhoA/Rho激酶(ROCK)信号通路对Toll样受体2(TLR-2)配体诱导的类风湿关节炎(RA)成纤维样滑膜细胞(FLS)分泌趋化因子的影响.方法 RA FLS来源于活动性RA患者滑膜组织;肽聚糖被用于TLR-2配体;RhoA活性检测采用pull down方法;ROCK活性用磷酸化肌球蛋白结合亚单位1(MYPT1)蛋白表达来表示,磷酸化MYPT1蛋白检测采用免疫印迹法;细胞活性检测采用四甲基偶氮唑蓝比色(MTT)法,趋化因子水平采用酶联免疫吸附测定法(ELISA)检测.结果 肽聚糖(5 mg/L)刺激使体外培养的RA FLS分泌白细胞介素8(IL-8)、单核细胞趋化蛋白-2(MCP-2)和受激活调节正常T细胞表达和分泌因子(RANTES)明显增高,对RhoA和ROCK活化也有显著的刺激作用,肽聚糖诱导的上述作用能被TLR-2单克隆抗体所阻抑.RhoA抑制剂C3转化酶和ROCK抑制剂Y27632对PG诱导的IL-8、MCP-2和RANTES等趋化因子分泌有显著的抑制作用.结论 RhoA/ROCK信号通路对TLR2介导的RA FLS分泌趋化因子具有调控作用,通过抑制该通路活化可能有利于RA的治疗.

Abstract：

Objective To evaluate the modulation of RhoA/Rho kinase (ROCK) signaling pathway,a small Rho GTPase that is considered as an important modulator in inflammatory responses,on Toll-like receptor-2 mediated chemokine secretion in fibroblast-like synoviocytes (FLS)from rheumatoid arthritis (RA) patients.Methods The RhoA activity was measured by a pull-down assay.And the ROCK activity was assessed by Western blot.The secretion of chemokines was measured by ELISA (enzyme-linked immunosorbent assay).MTT test was used to detect the cellular viability.Results The stimulation of peptidoglycan(PG,5 mg/L) increased the levels of IL-8 (interleukin-8),RANTES (regulated upon activation normal T cell expressed & secreted) and MCP-2 (monocyte chemotactic protein-2)and boosted the activities of RhoA and ROCK versus the unstimulated RA FLS.And these effects of PG were suppressed by anti-TLR-2 monoclonal antibody.Inhibition of RhoA and ROCK with a specific inhibitor inhibited the secretion of IL-8,RANTES and MCP-2 in PG-induced RA FLS.Conclusion The present study provides novel evidence that the RhoA/ROCK signal pathway modulates the TLR-2-mediated secretion of chemokines in RA FLS.It suggests that the inhibition of RhoA/ROCK may be a new therapeutic approach for RA.     

多发性硬化与趋化因子及其受体研究进展

多发性硬化是一种病因不明的中枢神经系统慢性脱髓鞘性炎症性疾病。趋化因子是一类具有趋化功能的细胞因子,在向炎症区募集并激活白细胞中发挥着主要作用,其与多发性性硬化的发生、发展与转归关系密切,将为揭示多发性硬化的发病机制、寻找诊断和治疗多发性硬化的新靶点提供新的契机。本文将对趋化因子及受体的分类、特性、致炎机制,及其与多发性硬化(MS)的关系进行阐述。