RhoA、ROCK在乳腺癌组织中的表达及其意义

目的：研究RhoA、ROCK在乳腺癌组织中的表达,探讨其与乳腺癌转移的相关性。方法：应用免疫组化S-P法,检测30例正常乳腺、58例乳腺癌组织中RhoA、ROCK蛋白的表达情况。分析其表达结果与临床病理特征的关系。结果：①RhoA、ROCK蛋白在正常乳腺组织中的表达率分别为0%、0%,在乳腺癌组织中的表达率分别为70.7%、81.0%,两者在乳腺癌组织中的表达水平明显高于正常乳腺组织（P〈0.05）。②RhoA、ROCK蛋白表达与患者年龄、组织学分级无关（P〉0.05）,与肿瘤大小、临床分期及有无淋巴结转移有关。③RhoA、ROCK蛋白两者表达水平呈正相关关系（P〈0.05）。结论：RhoA、ROCK蛋白的表达与乳腺癌转移及预后有一定关系。结论：RhoA、ROCK的表达对评估乳腺癌的转移和预后具有一定的临床价值。     

RhoA/ROCK信号通路在动脉粥样硬化发生中的作用

RhoA/ROCK通路参与细胞肌动蛋白骨架的调节、细胞的收缩、黏附、增生、分化及基因的表达等.越来越多的证据表明,其在动脉粥样硬化的发生发展中起着重要作用.对RhoA/ROCK通路的研究将拓展对有关心血管疾病的认识,有望为心血管疾病提供新的治疗靶点.     

溶血磷脂酸调控RhoA/ROCK2信号通路对乳腺癌细胞增殖的影响

目的 探讨溶血磷脂酸(LPA)与RhoA/ROCK2信号通路对乳腺癌细胞增殖的影响及其作用机制.方法 以不同浓度LPA干预乳腺癌MDA-MB-231细胞,每隔24 h以细胞计数法观察和记录细胞的增殖.以最佳LPA促增殖浓度作用于MDA-MB-231细胞,观察Rho激酶抑制剂(Y-27632)对癌细胞的影响;以Pull-down及Western blot法检测各组细胞内RhoA活性及RhoA、ROCK2蛋白表达.结果 LPA以时间及剂量依赖性关系显著促进MDA-MB-231细胞的增殖(P＜0.05);Y-27632可以显著抑制LPA的促增殖作用;LPA干预后RhoA活性及RhoA、ROCK2蛋白表达显著升高(P＜0.05),Y-27632干预后RhoA活性及RhoA、ROCK2蛋白表达显著下降(P＜0.05).结论 LPA可能通过调控RhoA/ROCK2信号通路促进乳腺癌细胞的增殖,为乳腺癌的临床治疗提供了新思路.     

早期骨关节炎软骨下骨趋化因子信号通路的表达

背景：趋化因子广泛存在于炎症反应组织当中,起诱导炎症细胞趋向炎症组织参与免疫应答的作用,大量的实验研究表明,趋化因子在骨关节炎致病过程中起了重要作用。 目的：分析早期骨关节炎软骨下骨的趋化因子信号通路表达改变情况,为阐述其在骨关节炎致病机制中的重要作用提供依据。 方法：30只SD大鼠随机均分为实验组和对照组。实验组切除右膝内侧半月板及内侧副韧带,对照组仅切开关节囊。于造模后1,2,4周取右膝关节标本,采用全基因表达谱芯片技术研究软骨下骨全基因表达,利用差异基因分析、信号通路分析的方法分析趋化因子信号通路的表达改变情况。 结果与结论：①发现一系列与趋化因子信号通路相关的差异表达基因。②实验组与对照组相比较,趋化因子信号通路在造模后1周差异有显著性意义(P < 0.05),造模后2,4周差异无显著性意义(P > 0.05)。说明趋化因子信号通路在极早期膝关节骨关节炎的软骨下骨改变中起重要作用。     

益母草碱调节RhoA/ROCK信号通路对多囊卵巢综合征大鼠卵巢颗粒细胞自噬和凋亡的影响

目的 探讨益母草碱（Leo）调节RhoA/ROCK信号通路对多囊卵巢综合征（PCOS）大鼠卵巢颗粒细胞（GCs自噬和凋亡的影响。方法 将SD大鼠随机分为正常组（NC组）和模型组（PCOS组）,分离卵巢GCs,H-E染色观察其形态;FSHR免疫荧光鉴定PCOS大鼠卵巢GCs。不同浓度的Leo处理PCOS大鼠卵巢GCs,CCK-8法检测细胞增殖并筛选最佳药物浓度。将PCOS大鼠GCs随机分为模型组（PCOS组）、益母草碱低剂量组（Leo-L组）、益母草碱中剂量组（Leo-M组）、益母草碱高剂量组（Leo-H组）、益母草碱高剂量组+RhoA/ROCK激活剂组（Leo+LPA组）,流式细胞术检测细胞凋亡率;透射电子显微镜检测细胞自噬;Western blot检测Bax、Cleaved-Caspase-3、Bcl-2、Beclin1、LC3-Ⅱ、LC3-Ⅰ、RhoA、ROCK1、ROCK2蛋白表达。结果 与NC组比较,PCOS组可见大小不等的囊性卵泡,且卵巢GCs数增多（P<0.05）;PCOS大鼠FSHR阳性率达92.13%,即成功分离卵巢GCs;Leo处理PCOS卵巢GCs后,细胞凋亡...     

RhoA和ROCK2在SOD1-G93A转基因小鼠脊髓中的表达

目的:通过研究Rho A和ROCK2在SOD1-G93A转基因小鼠脊髓内的表达变化以阐明Rho/ROCK信号通路在肌萎缩侧索硬化症(ALS)病程中的作用。方法:饲养SOD1-G93A转基因小鼠和同窝野生型小鼠至发病早期、中期和晚期,部分小鼠冰上剥离新鲜脊髓组织,利用RT-PCR方法检测Rho A和ROCK2 mRNA的表达,利用Western Blot方法检测Rho A和ROCK2蛋白的表达;部分小鼠行心脏灌注并剥离其脊髓组织制成冰冻切片,利用免疫组织化学染色方法检测Rho A和ROCK2蛋白的表达。结果:在SOD1-G93A鼠发病的早期、中期和晚期,转基因小鼠脊髓中Rho A和ROCK2的mRNA及蛋白表达均上调。免疫组织化学染色实验结果显示,野生型小鼠脊髓中Rho A和ROCK2弥散分布于胞质和突起中,阳性染色浅,SOD1-G93A转基因小鼠脊髓中Rho A和ROCK2阳性染色深,大量聚集在细胞膜及细胞质。结论:Rho A和ROCK2在SOD1-G93A转基因小鼠脊髓中异常高水平表达与ALS脊髓区病变密切相关,可能参与ALS疾病进程。     

RhoA/ROCKI信号通路在紫杉醇诱导宫颈癌细胞周期阻滞中的作用

目的 探讨RhoA/ROCKI信号通路在紫杉醇诱导宫颈癌细胞周期阻滞中的作用及其分子机制。方法 MTT法确定能引起细胞生长显著性抑制最小浓度 (1 μmol/L) 的紫杉醇。流式细胞仪检测细胞周期和RhoA蛋白表达,Western blotting检测ROCKI、细胞周期蛋白 (cyclins) 及周期依赖性激酶 (CDKs) 的表达。结果 紫杉醇处理C33A细胞后,G₂/M细胞数和RhoA/ROCKI蛋白表达显著性增加,并呈时间依赖性;而cyclin B1、A、D1和CDK1蛋白表达显著性降低。上述现象被RhoA抑制剂C3转移酶全部反转;却被ROCKI蛋白抑制剂Y-27632部分反转,但与对照相比差异仍显著。紫杉醇或RhoA/ROCKI抑制剂对CDK2的表达无显著性影响。结论 紫杉醇诱导了宫颈鳞癌C33A细胞的G₂/M期阻滞,其机制与RhoA活性增强后抑制特异的cyclins和CDKs有关,而下游通路中ROCKI活性仅起到部分作用。     

骨髓间充质干细胞向软骨及骨分化：Wnt5a/PCP信号通路作用的研究与进展

BACKGROUND: Wnt5a is able to inhibit canonical Wnt signaling and activate non-canonical Wnt signaling pathway. In recent years, it has been found that non-classical Wnt5a/PCP signaling pathway mediated by Wnt5a plays an important role in the process of bone marrow mesenchymal stem cell proliferation and differentiation, but the underlying mechanism is unclear. OBJECTIVE: To summarize the progress in downstream effector molecules related to Wnt5a/PCP signaling pathway, and its roles in the chondrogenic and osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: A computer-based online search of CqVip, CNKI and PubMed databases between January 2000 and February 2016 was performed using the Chinese keywords of “BMSCs, Wnt signaling pathways, chondrogenic differentiation, osteogenic differentiation” and English keywords of “BMSCs, chondrogenic differentiation, osteogenic differentiation, Wnt, Fzd, Ror2, RhoA, ROCK, JNK”, respectively. Literatures related to bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation were selected. Finally, 43 eligible articles were included for analysis through excluding the old and repeated research. RESULTS AND CONCLUSION:Wnt5a, a representative protein in non-canonical Wnt signaling pathway, paticipates in the cytoskeleton, cell migration and cell polarization and other activities by mediating its downstream signaling molecules such as Fzd, Ror, RhoA, ROCK, JNK, thereby regulating its proliferation and differentiation. But it is unclear how Wnt5a/PCP participates in the bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation and how the downstream effector molecules interact or function independently, which requires further studies. 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     

Ezrin蛋白在软骨肉瘤中的表达及其意义

目的 探讨Ezrin蛋白在软骨肉瘤中的表达状况及其与相关临床病理因素的关系,以及可能的临床意义.方法 采用鼠抗人Ezrin单克隆抗体和免疫组化MaxVision法,检测42例软骨肉瘤和20例软骨瘤中Ezrin蛋白的表达.结果 (1)Ezrin在软骨肉瘤组和软骨瘤组中的阳性表达率分别为61.90%和25.00%(P<0.05).(2)Ezrin在普通型、透明细胞型和去分化型软骨肉瘤中的阳性表达率分别为60.61%、50%、71.43%.(3)Ezrin在普通型软骨肉瘤的Ⅰ-Ⅲ级中的阳性表达率分别为25%、64.29%、81.82%(P<0.05).结论 膜细胞骨架连接蛋白Ezrin在软骨肉瘤中高表达,提示可能参与软骨肉瘤的侵袭和转移;检测Ezrin蛋白可以辅助鉴别诊断软骨肉瘤与增生活跃之软骨瘤.     

信号通路在膝骨关节炎实验研究中的进展

BACKGROUND: Studies have shown that signaling pathway plays an important role in the pathogenesis of knee osteoarthritis. OBJECTIVE: To elaborate the experimental progress of signal transduction pathway in knee osteoarthritis METHODS: A computer-based retrieval of PubMed, CNKI, VIP and Wanfang databases was performed to find the relevant literatures concerning experimental studies of signaling pathway in knee osteoarthritis. All data were primarily screened to exclude repeated and irrelevant articles. Articles about association between knee osteoarthritis and signal pathway were included.   RESULTS AND CONCLUSION: A total of 51 articles were collected, including 20 in Chinese and 31 in English. It is concluded that the signal transduction pathway related to knee osteoarthritis is mainly concentrated in the two aspects: the proliferation of chondrocytes and the apoptosis of chondrocytes. The signal transduction pathways related to the proliferation of chondrocytes mainly include Notch signaling pathway and Wnt signaling pathway. SDF-1/CXCR4 signaling pathway, TLR4 signaling pathway, and MAPKs signaling pathway are associated with apoptosis in chondrocytes. Hippo-YAP signaling pathway and ERK1/2 signaling pathway have a double role in the regulation of proliferation and apoptosis of chondrocytes.      

Involvement of RhoA/ROCK1 signaling pathway in hyperglycemia-induced microvascular endothelial dysfunction in diabetic retinopathy

Diabetic retinopathy (DR) is a well-known serious complication of diabetes mellitus (DM), and can eventually advance to end-stage blindness. In the early stage of DR, endothelial cell barrier disorganized primarily and tight junction (TJ) protein composition transformed subsequently. The small GTPase RhoA and its downstream effector Rho-associated coiled-coil containing protein kinase 1 (ROCK1) regulate a mass of cellular processes, including cell adherence, proliferation, permeability and apoptosis. Although RhoA inhibitors have provided substantial clinical benefit as hypertonicity therapeutics, their use is limited by complex microenvironment as DR. While ample evidence indicates that TJ can be influenced by the RhoA/ROCK1 signaling, the underlying mechanisms remain incompletely understood. Here, we have uncovered a significant signaling network involved in diabetic retinal microvascular endothelial dysfunction (RMVED). Our results indicated that the activation of RhoA/ROCK1 pathway due to high glucose played a key role in microvascular endothelial cell dysfunction (MVED) by way of directly inducing TJ proteins over-expression during DR. We demonstrated that inhibition of RhoA/ROCK1 may attenuate the hypertonicity of endothelial cell caused by high glucose microenvironment meanwhile. Besides, chemical and pharmacological inhibitors of RhoA/ROCK1 pathway may partly block inflammation due to DR. Simultaneously, the apoptosis aroused by high glucose was also prevented considerably by fasudil, a kind of pharmacological inhibitor of RhoA/ROCK1 pathway. These findings indicate that RhoA/ROCK1 signaling directly modulates MVED, suggesting a novel therapeutic target for DR.     

维药买朱尼对关节软骨前炎性因子的影响

背景：骨关节炎病理过程中,白细胞介素1β被认为是促进软骨基质降解和关节软骨破坏的最重要的细胞因之一。 目的：观察白细胞介素1β在关节软骨中的表达,并观察维药买朱尼对其的影响。 方法：将40只SD大鼠随机数字表法随机等分为模型对照组、维药买朱尼组、假手术组、正常对照组。模型对照组和维药买朱尼组采用改良Hulth造模法建立大鼠膝骨关节炎模型,假手术组仅显露膝关节,不切断韧带,不切除内侧半月板。维药买朱尼组建模第2周开始灌胃维药买朱尼10.31 mg/(kg•d),模型组及假手术组大鼠均灌服等量生理盐水,连续4周。 结果与结论：模型组软骨退变程度明显重于维药买朱尼组,模型组软骨大体评分及Mankin评分均明显高于维药买朱尼组(P < 0.05),模型组软骨细胞白细胞介素1β的表达强度亦明显高于维药买朱尼组(P < 0.05)。与正常对照组比较,假手术组软骨大体评分、Mankin评分及软骨细胞白细胞介素1β差异无显著性意义 (P > 0.05)。结果说明,维药买朱尼可以抑制关节软骨前炎性因子白细胞介素1β的表达。     

雌激素与机械张应力对骨重建相关因子的作用

背景：牙槽骨是人体骨代谢最活跃的区域之一,对于在正畸治疗过程中的女性患者容易受到雌激素水平的影响。 目的：观察雌激素对机械张应力作用下的牙周膜成纤维细胞骨重建相关因子骨保护素、核因子κB受体活化子配体mRNA表达水平的影响。 方法：利用一定浓度雌激素干预人牙周膜成纤维细胞后使用四点弯曲加力装置对细胞进行不同时间段的机械张应力加载,并利用RT-PCR检测雌激素与机械张应力共同作用后牙周膜成纤维细胞骨保护素、核因子κB受体活化子配体基因表达变化。 结果与结论：雌激素刺激后再进行机械张应力加载的人牙周膜成纤维细胞跟单纯受到机械张应力作用的人牙周膜成纤维细胞相比,骨保护素mRNA表达量明显上调,而核因子κB受体活化子配体mRNA表达量显著下降。实验表明雌激素对机械张应力状态下的牙周膜成纤维细胞骨重建相关因子核因子κB受体活化子配体、骨保护素有较强调控作用。     

人膝骨性关节炎滑膜中表达的骨桥蛋白

背景：目前研究显示骨桥蛋白与骨性关节炎关节软骨退变密切相关,但骨桥蛋白与骨性关节炎滑膜病变是否相关,仍少见报道。 目的：研究骨桥蛋白在原发性骨性关节炎发生与发展过程中的作用。 方法：选取膝骨性关节炎患者关节滑膜标本与下肢外伤患者作对照,根据综合评分法进行分组,采用免疫组化方法检测关节滑膜中骨桥蛋白水平,比较不同程度骨性关节炎组膝关节滑膜中骨桥蛋白的差异,同时比较标本滑膜衬里层和衬里下层骨桥蛋白的水平。 结果与结论：骨性关节炎组织中骨桥蛋白阳性呈黄色、棕黄色、棕褐色表达,病变程度越重,颜色表达越深。骨性关节炎组滑膜中骨桥蛋白明显高于对照组,差异有显著性意义(P < 0.05),且随着骨性关节炎病情加重,骨桥蛋白的表达逐渐增多,呈正相关(ρ=0.663,P < 0.01)。但滑膜衬里层和衬里下层骨桥蛋白的水平差异无显著性意义(P > 0.05)。说明骨桥蛋白可能与骨性关节炎的发病有关。     

The effects of joint immobilization on articular cartilage of the knee in previously exercised rats

Studies have determined the effects of joint immobilization on the articular cartilage of sedentary animals, but we are not aware of any studies reporting the effects of joint immobilization in previously trained animals. The objective of the present study was to determine whether exercise could prevent degeneration of the articular cartilage that accompanies joint immobilization. We used light microscopy to study the thickness, cell density, nuclear size, and collagen density of articular cartilage of the femoral condyle of Wistar rats subjected to aerobic physical activity on an adapted treadmill five times per week. Four groups of Wistar rats were used: a control group (C), an immobilized group (I), an exercised group (E), and an exercised and then immobilized group (EI). The right knee joints from rats in groups I and EI were immobilized at 90 °C of flexion using a plastic cast for 8 weeks. Cartilage thickness decreased significantly in group I (mean, 120.14 ± 15.6 μm, P < 0.05), but not in group EI (mean, 174 ± 2.25), and increased significantly in group E (mean, 289.49 ± 9.15) compared with group C (mean, 239.20 ± 6.25). The same results were obtained for cell density, nuclear size, and collagen density (in all cases, P < 0.05). We concluded that exercise can prevent degenerative changes in femoral articular cartilage caused by immobilization of the knee joint.     

体外传代培养兔关节软骨细胞的去分化现象

背景：兔是软骨组织工程研究中应用非常广泛的实验动物模型,关节软骨细胞的去分化现象已经被广泛认可。 目的：观察体外传代培养过程中兔关节软骨细胞的去分化现象。 方法：将从新西兰大白兔膝关节分离获取的原代软骨细胞进行传代培养至第7代,对细胞生长、形态、基质分泌以及基因表达等方面分别采用细胞计数,显微镜观察,F机动蛋白染色、番红O染色,糖胺聚糖定量测定以及半定量聚合链式反应进行鉴定和比较。 结果与结论：光学显微镜下,兔关节软骨细胞在体外传代培养过程中细胞形态由小且圆形或多角形,逐步转变大且为成纤维细胞样的梭形形态,对F机动蛋白的染色进一步佐证了这样的形态变化。细胞计数结果表明,细胞增殖能力随代次增加显著下降,特别是第3代以后的软骨细胞基本无明显增殖;经过番红O染色以及定量测定糖胺聚糖的含量,发现软骨特性胞外基质分泌量从第2代细胞开始就呈现显著的降低。半定量聚合链式反应检测结果表明,随着传代次数的增加,特别是第3代以后,软骨相关特征分子(包括Ⅱ型胶原、聚集蛋白聚糖、软骨寡聚基质蛋白和SOX9等)基因表达水平下调;而与去分化相关的特征分子Ⅰ型胶原和多能蛋白聚糖基因表达水平上调,同时,细胞表面分子CD90基因表达上调,而CD14基因表达未见明显变化。结果证实,兔软骨细胞在体外传代过程中可出现快速地去分化现象,呈现出特征基因表达水平的变化,第3代以内的软骨细胞适合应用于软骨组织再生修复。     

抑制G蛋白偶联受体40通过RhoA/ROCK1信号通路缓解小鼠过敏性哮喘

目的:探究抑制G蛋白偶联受体40(GPR40)减轻过敏性哮喘小鼠症状的作用及机制.方法:将28只雄性C57BL/6小鼠随机分为正常对照组、哮喘模型组(用卵清蛋白建立过敏性哮喘模型)、低剂量GPR40抑制剂DC260126干预组(3 mg/kg DC260126组)和高剂量DC260126干预组(10 mg/kg DC2...     

Tensile stress patterns predicted in the articular disc of the human temporomandibular joint

The direction of the first principal stress in the articular disc of the temporomandibular joint was predicted with a biomechanical model of the human masticatory system. The results were compared with the orientation of its collagen fibers. Furthermore, the effect of an active pull of the superior lateral pterygoid muscle, which is directly attached to the articular disc, was studied. It was hypothesized that the markedly antero-posterior direction of the collagen fibers would be reflected in the direction of the tensile stresses in the disc and that active pull of the superior lateral pterygoid muscle would augment these tensions. It was found that the tensile patterns were extremely dependent on the stage of movement and on the mandibular position. They differed between the superior and inferior layers of the disc. The hypothesis could only be confirmed for the anterior and middle portions of the disc. The predicted tensile principal stresses in the posterior part of the disc alternated between antero-posterior and medio-lateral directions.