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Michael S Samuel, Nicola Rath, Siti F Masre, Sarah T Boyle, David A Greenhalgh, Marina Kochetkova, Sheila Bryson, David Stevenson, Michael F Olson
The serine/threonine kinases ROCK1 and ROCK2 are central mediators of actomyosin contractile force generation that act downstream of the RhoA small GTP-binding protein. As a result, they have key roles in regulating cell morphology and proliferation, and have been implicated in numerous pathological conditions and diseases including hypertension and cancer. Here we describe the generation of a gene-targeted mouse line that enables CRE-inducible expression of a conditionally-active fusion between the ROCK2 kinase domain and the hormone-binding domain of a mutated estrogen receptor (ROCK2:ER)...
October 24, 2016: Genesis: the Journal of Genetics and Development
Xiangbing Wu, Chandler L Walker, Qingbo Lu, Wei Wu, Daniel B Eddelman, Jonathan M Parish, Xiao-Ming Xu
Activation of RhoA/Rho kinase leads to growth cone collapse and neurite retraction. Although RhoA/Rho kinase inhibition has been shown to improve axon regeneration, remyelination and functional recovery, its role in neuronal cell death remains unclear. To determine whether RhoA/Rho kinase played a role in neuronal death after injury, we investigated the relationship between RhoA/Rho kinase and cytosolic phospholipase A2 (cPLA2), a lipase that mediates inflammation and cell death, using an in vitro neuronal death model and an in vivo contusive spinal cord injury model performed at the 10th thoracic (T10) vertebral level...
October 22, 2016: Molecular Neurobiology
Binod Kumar, Dipanjan Dutta, Jawed Iqbal, Mairaj Ahmed Ansari, Arunava Roy, Leela Chikoti, Gina Pisano, Mohanan Valiya Veettil, Bala Chandran
Kaposi's sarcoma-associated herpesvirus (KSHV) binding to the endothelial cell surface heparan sulfate is followed by sequential interactions with α3β1, αVβ3 and αVβ5 integrins and Ephrin A2 receptor tyrosine kinase (EphA2R). These interactions activate host cell pre-existing FAK, Src, PI3-K and RhoGTPase signaling cascades, c-Cbl mediated ubiquitination of receptors, recruitment of CIB1, p130Cas and Crk adaptor molecules, and membrane bleb formation leading to lipid raft dependent macropinocytosis of KSHV into human microvascular dermal endothelial (HMVEC-d) cells...
October 2016: PLoS Pathogens
Pieter Uvin, Maarten Albersen, Ine Bollen, Maarten Falter, Emmanuel Weyne, Loes Linsen, Hanna Tinel, Peter Sandner, Trinity J Bivalacqua, Dirk Jmk De Ridder, Frank Van der Aa, Bert Brône, Koenraad Van Renterghem
OBJECTIVES: To evaluate the expression of the Rho/Rho associated protein kinase (ROCK) pathway in corpus cavernosum of patients with severe erectile dysfunction (ED) compared to healthy human corpus cavernosum, and to test the functional effects of two Rho Kinase Inhibitors (RKI) on erectile tissue of patients with severe ED, not responding to phosphodiesterase type 5 inhibitors (PDE5-i). PATIENTS AND METHODS: Human corpus cavernosum samples were obtained after consent from individuals undergoing penile prosthesis implantation (n = 7 for organ bath experiments, n = 17 for qPCR)...
October 20, 2016: BJU International
Anping Cai, Yingqing Feng, Yingling Zhou, Jiyan Chen
OBJECTIVE: To investigate the effects and mechanisms of levorotatory (L)-amlodipine combined fluvastatin treatment on endothelium-mesenchymal transition (EndMT), a process related to hypertension development. DESIGN AND METHOD: Human umbilical vein endothelial cells (HUVECs) were used and divided to six groups based on different therapeutic strategies as follows: Blank Control, angiotensin-II (Ang-II), Ang-II+ rho-associated kinase (ROCK) inhibitor, Ang-II+ L-amlodipine, Ang-II+ fluvastatin, and Ang-II +L-amlodipine + fluvastatin groups...
September 2016: Journal of Hypertension
Kangwook Lee, Sung-Gook Cho, Sang-Mi Woo, Ah Jeong Kim, Kang Min Lee, Ho Yeon Go, Seung-Ho Sun, Tae-Hun Kim, Ki-Yong Jung, You-Kyung Choi, Eun Mee Lim, Yun-Kyung Song, Jong-Hyeong Park, Chan-Yong Jun, Seong-Gyu Ko
Danggui-Sayuk-Ga-Osuyu-Senggang-Tang (DSGOST), one of the traditional Chinese medicines, has long been prescribed for patients suffering from Raynaud phenomenon (RP) in Northeast Asian countries, including China, Japan and Korea. Although a previous in vitro study from our laboratory revealed that DSGOST prevents cold (25˚C)‑induced RhoA activation and endothelin‑1 (ET‑1) production in endothelial cells (ECs), the mechanisms by which DSGOST is able to alleviate the symptoms of RP have yet to be fully elucidated...
October 5, 2016: Molecular Medicine Reports
Zhijun Sun, Xing Wu, Weiping Li, Hui Peng, Xuhua Shen, Lu Ma, Huirong Liu, Hongwei Li
BACKGROUND: Diabetes-induced vascular dysfunction may arise from reduced nitric oxide (NO) availability, following interaction with superoxide to form peroxynitrite. Peroxynitrite can induce formation of 3-nitrotyrosine-modified proteins. RhoA/ROCK signaling is also involved in diabetes-induced vascular dysfunction. The study aimed to investigate possible links between Rho/ROCK signaling, hyperglycemia, and peroxynitrite in small coronary arteries. METHODS: Rat small coronary arteries were exposed to normal (NG; 5...
October 11, 2016: BMC Cardiovascular Disorders
Ko-Chao Lee, Hsing-Chun Kuo, Chien-Heng Shen, Chien-Chang Lu, Wen-Shih Huang, Meng-Chiao Hsieh, Cheng-Yi Huang, Yi-Hung Kuo, Yung-Yu Hsieh, Chih-Chuan Teng, Li-Ya Lee, Shui-Yi Tung
Erinacine A, a major active component of a diterpenoid derivative isolated from Hericium erinaceus mycelium, has been demonstrated to exert anticancer effects. Herein, we present an investigation of the molecular mechanism of erinacine A induction associated with cancer cells' aggressive status and death. A proteomic approach was used to purify and identify the differentially expressed proteins following erinacine A treatment and the mechanism of its action in apoptotic and the targets of erinacine A. Our results demonstrate that erinacine A treatment of HCT-116 and DLD-1 cells increased cell cytotoxicity and reactive oxygen species (ROS) production as well as decreased cell proliferation and invasiveness...
October 6, 2016: Journal of Cellular and Molecular Medicine
L C Wu, C Liu, M R Jiang, Y M Jiang, Q H Wang, Z Y Lu, S J Wang, W L Yang, Y X Shao
Development of the eyelid requires coordination of the cellular processes involved in proliferation, cell size alteration, migration, and cell death. C57BL/6J-corneal opacity (B6-Co) mice are mutant mice generated by the administration of N-ethyl-N-nitrosourea (100 mg/kg). They exhibit the eyelids open at birth phenotype, abnormal round cell shape from tightened F-actin bundles in leading edge keratinocytes at E16.5, and gradual corneal opacity with neovessels. The tip of the leading edge in B6-Co mice did not move forward, and demonstrated a sharp peak shape without obvious directionality...
August 26, 2016: Genetics and Molecular Research: GMR
Kai Cui, Yang Luan, Tao Wang, Li Zhuan, Ke Rao, Shao-Gang Wang, Zhang-Qun Ye, Ji-Hong Liu, Dao-Wen Wang
Our previous studies have demonstrated that erectile function was preserved in aged transgenic rats (TGR) harboring the human tissue kallikrein 1 (hKLK1), while the molecular level of hKLK1 on corporal fibrosis to inhibit age-related erectile dysfunction (ED) is poorly understood. Male wild-type Sprague-Dawley rats (WTR) and TGR harboring the hKLK1 gene were fed to 4- or 18-month-old and divided into three groups: young WTR (yWTR) as the control, aged WTR (aWTR), and aged TGR (aTGR). Erectile function of all rats was assessed by cavernous nerve electrostimulation method...
September 27, 2016: Asian Journal of Andrology
Shugang Xu, Xing Guo, Xiao Gao, Hao Xue, Jinsen Zhang, Xiaofan Guo, Wei Qiu, Ping Zhang, Gang Li
Macrophage migration inhibitory factor (MIF) is highly expressed in glioblastoma, promoting malignant progression and suppresses immune surveillance. However, the mechanism underlying its biological roles in human glioblastoma and the capability of MIF to escape dendritic cell (DC) surveillance remain poorly understood. In the present study, we found that recombinant human MIF (rhMIF) activated the RhoA‑ROCK1 pathway and simultaneously upregulated F‑actin fibre formation. Additionally, we showed that rhMIF increased autophagy in glioblastoma cells, and knockdown of endogenic MIF suppressed autophagy...
September 23, 2016: International Journal of Oncology
Anping Cai, Yingqing Feng, Yingling Zhou, Jiyan Chen
OBJECTIVE: To investigate the effects and mechanisms of levorotatory (L)-amlodipine combined fluvastatin treatment on endothelium-mesenchymal transition (EndMT), a process related to hypertension development. DESIGN AND METHOD: Human umbilical vein endothelial cells (HUVECs) were used and divided to six groups based on different therapeutic strategies as follows: Blank Control, angiotensin-II (Ang-II), Ang-II+ rho-associated kinase (ROCK) inhibitor, Ang-II+ L-amlodipine, Ang-II+ fluvastatin, and Ang-II +L-amlodipine + fluvastatin groups...
September 2016: Journal of Hypertension
Jiong Yue, Wei Li, Chao Liang, Bing Chen, Xin Chen, Lukang Wang, Zhenle Zang, Sixun Yu, Shiyong Liu, Song Li, Hui Yang
Temporal lobe epilepsy (TLE) is a frequent form of focal intractable epilepsy in adults, but the specific mechanism underlying the epileptogenesis of TLE is still unknown. Human leukocyte immunoglobulin-like receptor B2 (LILRB2) (the murine homolog gene called paired immunoglobulin-like receptor B, or PirB), participates in the process of synaptic plasticity and neurite growth in the central nervous system (CNS), suggesting a potential role of LILRB2 in epilepsy. However, the expression pattern of LILRB2 and the downstream molecular signal in intractable TLE remains poorly understood...
November 2016: Experimental Neurology
Sonia I Vlaicu, Alexandru Tatomir, Dallas Boodhoo, Takahiro Ito, Matthew Fosbrink, Cornelia Cudrici, Armugam P Mekala, Jonathan Ciriello, Doiniţa Crişan, Emil Boţan, Violeta Rus, Horea Rus
The complement system is an important player in the development of atherosclerosis. Previously reported as a cell cycle regulator, RGC-32 is an essential effector of the terminal complement complex, C5b-9. In this study, our aims were to determine the expression of RGC-32 in the human atherosclerotic arterial wall and to delineate the mechanisms through which RGC-32 affects C5b-9-induced endothelial cell proliferation and migration. We now demonstrate that RGC-32 is expressed in human aortic atherosclerotic wall and that RGC-32 expression increases with the progression of atherosclerosis...
September 13, 2016: Experimental and Molecular Pathology
Li-Juan Liu, Feng-Juan Yao, Gui-Hua Lu, Cheng-Gui Xu, Zhe Xu, Kai Tang, Yun-Jiu Cheng, Xiu-Ren Gao, Su-Hua Wu
OBJECTIVES: To study the role of the Rho/ROCK pathway in Ang II and TGF-β1-induced atrial remodeling. METHODS AND RESULTS: A canine atrial fibrillation (AF) model was established by rapid atrial pacing (RAP) of the left atrium. The roles of TGF-β1, the RhoA/ROCK signaling pathway and connective tissue growth factor (CTGF) in atrial remodeling were studied via both in vitro and in vivo experiments. Each of the dogs that received RAP developed persistent AF within 4 weeks...
2016: PloS One
Lei Zhang, Guoliang Jiang, Xueling Zhao, Yuekun Gong
BACKGROUND/AIMS: We investigated the role of dimethyloxalylglycine (DMOG) in bone marrow mesenchymal stem cell (BMSC) osteogenesis mediated by RhoA/ROCK. METHODS: BMSCs were cultured with and without DMOG and/or Y-27632 (ROCK1 inhibitor). Cell proliferation, alkaline phosphatase (ALP) levels, and calcium deposits were determined. The expression of Runx2, OSX, p-cofilin, RhoA, and GTP-bound RhoA was determined by real-time RT-PCR and Western blot. Rho-associated coiled-coil-containing protein kinase (ROCK) activity was determined by measuring the phosphorylation of myosin-binding subunit of myosin phosphatase using an ELISA kit...
2016: Cellular Physiology and Biochemistry
Min Shi, Jue Wei, Wen-Ying Meng, Na Wang, Ting Wang, Yu-Gang Wang
The current study investigated the effects of phased joint intervention on clinical efficacy and Rho/Rho-associated coil protein kinase (ROCK) expression in patients with portal hypertension complicated by esophageal variceal bleeding (EVB) and hypersplenism. Patients with portal hypertension (n=53) caused by liver cirrhosis complicated by EVB and hypersplenism treated with phased joint intervention were assessed, and portal hemodynamics, blood, liver function, complications, and rebleeding incidence were analyzed...
September 2016: Experimental and Therapeutic Medicine
Caroline Gastebois, Stéphanie Chanon, Sophie Rome, Christine Durand, Elise Pelascini, Audrey Jalabert, Vanessa Euthine, Vincent Pialoux, Stéphane Blanc, Chantal Simon, Etienne Lefai
This study investigated miR-148b as a potential physiological actor of physical inactivity-induced effects in skeletal muscle. By using animal and human protocols, we demonstrated that the early phase of transition toward inactivity was associated with an increase in muscle miR-148b content, which triggered the downregulation of NRAS and ROCK1 target genes. Using human myotubes, we demonstrated that overexpression of miR-148b decreased NRAS and ROCK1 protein levels, and PKB phosphorylation and glucose uptake in response to insulin...
September 2016: Physiological Reports
Nirpesh Adhikari, Sanjiv Neupane, Gi-Jeong Gwon, Ji-Youn Kim, Chang-Hyeon An, Sanggyu Lee, Wern-Joo Sohn, Youngkyun Lee, Jae-Young Kim
Grainyhead-like 3 (Grhl3) is a transcription factor involved in epithelial morphogenesis. In the present study, we evaluated the developmental role of Grhl3 in structural formation of the circumvallate papilla (CVP), which undergoes dynamic morphological changes during organogenesis. The specific expression pattern of Grhl3 was examined in the CVP-forming region, specifically in the apex and epithelial stalk from E13.5 to E15.5 using in situ hybridization. To determine the role of Grhl3 in epithelial morphogenesis of the CVP, we employed an in vitro tongue culture method, wherein E13...
September 1, 2016: Histochemistry and Cell Biology
Isabel Hinsenkamp, Sandra Schulz, Mareike Roscher, Anne-Maria Suhr, Björn Meyer, Bogdan Munteanu, Jens Fuchser, Stefan O Schoenberg, Matthias P A Ebert, Björn Wängler, Carsten Hopf, Elke Burgermeister
Gastric cancer (GC) remains a malignant disease with high mortality. Patients are frequently diagnosed in advanced stages where survival prognosis is poor. Thus, there is high medical need to find novel drug targets and treatment strategies. Recently, the comprehensive molecular characterization of GC subtypes revealed mutations in the small GTPase RHOA as a hallmark of diffuse-type GC. RHOA activates RHO-associated protein kinases (ROCK1/2) which regulate cell contractility, migration and growth and thus may play a role in cancer...
August 2016: Neoplasia: An International Journal for Oncology Research
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