Fasudil, a rho kinase inhibitor, limits motor neuron loss in experimental models of amyotrophic lateral sclerosis
Masashi TakataHirotaka TanakaMasahiko KimuraYuki NagaharaY. TanakaKoh KawasakiM SetoKazuhiro TsurumaMasamitsu ShimazawaHideaki Hara
114
Citation
29
Reference
10
Related Paper
Citation Trend
Abstract:
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder with no effective treatment. Fasudil hydrochloride (fasudil), a potent rho kinase (ROCK) inhibitor, is useful for the treatment of ischaemic diseases. In previous reports, fasudil improved pathology in mouse models of Alzheimer's disease and spinal muscular atrophy, but there is no evidence in that it can affect ALS. We therefore investigated its effects on experimental models of ALS.In mice motor neuron (NSC34) cells, the neuroprotective effect of hydroxyfasudil (M3), an active metabolite of fasudil, and its mechanism were evaluated. Moreover, the effects of fasudil, 30 and 100 mg·kg(-1), administered via drinking water to mutant superoxide dismutase 1 (SOD1(G93A)) mice were tested by measuring motor performance, survival time and histological changes, and its mechanism investigated.M3 prevented motor neuron cell death induced by SOD1(G93A). Furthermore, M3 suppressed both the increase in ROCK activity and phosphorylated phosphatase and tensin homologue deleted on chromosome 10 (PTEN), and the reduction in phosphorylated Akt induced by SOD1(G93A). These effects of M3 were attenuated by treatment with a PI3K inhibitor (LY294002). Moreover, fasudil slowed disease progression, increased survival time and reduced motor neuron loss, in SOD1(G93A) mice. Fasudil also attenuated the increase in ROCK activity and PTEN, and the reduction in Akt in SOD1(G93A) mice.These findings indicate that fasudil may be effective at suppressing motor neuron degeneration and symptom progression in ALS. Hence, fasudil may have potential as a therapeutic agent for ALS treatment.Keywords:
Fasudil
Rho kinase inhibitor
Objective Accumulating evidence suggests that inflammatory cell infiltration is crucial pathogenesis during the initiation and progression of abdominal aortic aneurysm (AAA). Given Rho-kinase (ROCK), an important kinase control the actin cytoskeleton, regulates the inflammatory cell infiltration, thus, we investigate the possibility and mechanism of preventing experimental AAA progression via targeting ROCK in mice porcine pancreatic elastase (PPE) model. Methods and Results AAA was created in 10-week-old male C57BL/6 mice by transient intraluminal porcine pancreatic elastase infusion into the infrarenal aorta. The mRNA level of RhoA, RhoC, ROCK1 and ROCK2 were elevated in aneurismal aorta. Next, PPE infusion mice were orally administrated with vehicle or ROCK inhibitor (Fasudil at dose of 200 mg/kg/day) during the period of day 1 prior to PPE infusion to day 14 after PPE infusion. PPE infusion mice treated with Fasudil produced significantly smaller aneurysms as compare to PPE infusion mice treated with vehicle. AAAs developed in all vehicle-treated groups within 14 days, whereas AAAs developed in six mice (66%, 6/9) treated with Fasudil within 14 days. Furthermore, our semi-quantitative histological analysis revealed that blood vessels and macrophages were significantly reduced in Fasudil treated mice during the AAA progression. Finally, when mice with existing AAAs were treated with Fasudil, the enlargement was nearly completely suppressed. Conclusion Fasudil inhibits experimental AAA progression and stabilize existing aneurysms, through mechanisms likely related to impaired mural macrophage infiltration and angiogenesis. These findings suggest that ROCK inhibitor may hold substantial translational value for AAA diseases.
Fasudil
ROCK2
Rho kinase inhibitor
Pancreatic elastase
Cite
Citations (2)
Cardiac allograft vasculopathy (CAV) continues to be a major cause of late graft failure after cardiac transplantation. We have demonstrated that Rho-kinase, an effector of the small GTPase Rho, plays an important role in the pathogenesis of arteriosclerosis. In this study, we examined whether the Rho-kinase–mediated pathway is also involved in the pathogenesis of CAV using a specific Rho-kinase inhibitor and a dominant-negative Rho-kinase. Hearts from AKR mice were heterotopically transplanted to C3H/He (allograft) or AKR mice (isograft), and the effects of long-term oral treatment with fasudil, which is metabolized to a specific Rho-kinase inhibitor hydroxyfasudil, on CAV were examined at 2 and 4 weeks after the transplantation. Coronary remodeling in the allografts characterized by intimal thickening and perivascular fibrosis was dose-dependently suppressed in the fasudil group compared with the control group ( P <0.01, n=9 to 10). The inhibitory effects of hydroxyfasudil were mimicked by in vivo gene transfer of dominant-negative Rho-kinase ( P <0.05, n=4). Among the proinflammatory cytokines examined, those of macrophage migration inhibitory factor, interferon-γ, and transforming growth factor-β1 were upregulated in the control group and were dose-dependently inhibited in the fasudil group ( P <0.01, n=5). Vascular inflammation in the allografts, as evidenced by accumulation of inflammatory cells (macrophages and T cells), was also significantly inhibited in the fasudil group ( P <0.05, n=5 to 10). These results indicate that long-term treatment with fasudil suppresses CAV in mice, suggesting that Rho-kinase is an important therapeutic target for the prevention of CAV.
Fasudil
Rho kinase inhibitor
Pathogenesis
Proinflammatory cytokine
Cite
Citations (85)
Rationale: The complex and multifactorial pathogenesis of pulmonary hypertension (PH) involves constriction, remodeling, and in situ thrombosis of pulmonary vessels. Both serotonin (5-HT) and Rho kinase signaling may contribute to these alterations.Objectives: To investigate possible links between the 5-HT transporter (5-HTT) and RhoA/Rho kinase pathways, as well as their involvement in the progression of human and experimental PH.Methods: Biochemical and functional analyses of lungs, platelets, and pulmonary artery smooth muscle cells (PA-SMCs) from patients with idiopathic PH (iPH) and 5-HTT overexpressing mice.Measurements and Main Results: Lungs, platelets, and PA-SMCs from patients with iPH were characterized by marked elevation in RhoA and Rho kinase activities and a strong increase in 5-HT binding to RhoA indicating RhoA serotonylation. The 5-HTT inhibitor fluoxetine and the type 2 transglutaminase inhibitor monodansylcadaverin prevented 5-HT–induced RhoA serotonylation and RhoA/Rho kinase activation, as well as 5-HT–induced proliferation of PA-SMCs from iPH patients that was also inhibited by the Rho kinase inhibitor fasudil. Increased Rho kinase activity, RhoA activation, and RhoA serotonylation were also observed in lungs from SM22–5-HTT+mice, which overexpress 5-HTT in smooth muscle and spontaneously develop PH. Treatment of SM22–5-HTT+ mice with either fasudil or fluoxetine limited PH progression and RhoA/Rho kinase activation.Conclusions: RhoA and Rho kinase activities are increased in iPH, in association with enhanced RhoA serotonylation. Direct involvement of the 5-HTT/RhoA/Rho kinase signaling pathway in 5-HT–mediated PA-SMC proliferation and platelet activation during PH progression identify RhoA/Rho kinase signaling as a promising target for new treatments against PH.
Fasudil
Rho kinase inhibitor
Cite
Citations (163)
We evaluated prevention of transforming growth factor β (TGFβ)-induced transdifferentiation of cultured scleral fibroblasts to myofibroblasts by rho-associated protein kinase (ROCK) inhibitors. Additionally, we tested whether local delivery of ROCK inhibitors reduced scleral fibroblast proliferation in response to chronic intraocular pressure (IOP) elevation.Primary human peripapillary sclera (PPS) fibroblasts were cultured and treated with TGFβ to induce myofibroblast transdifferentiation, as determined by immunoblot assessment of α smooth muscle actin (SMA) levels and collagen gel contraction. Cells were treated with the ROCK inhibitors Y27632, fasudil, and H1152 before TGFβ treatment. ROCK activity in TGFβ-treated fibroblasts and sclera from ocular hypertensive mice was assessed by measuring phosphorylation of the ROCK substrate MYPT1 at Thr696. Fibroblast proliferation following IOP elevation and ROCK inhibitor treatment was assessed by an enzyme-linked immunosorbent (ELISA) assay.ROCK inhibitors H1152 (10μM), Y27632 (10 μM), and fasudil (5μM) reduced SMA expression 72%, 85%, and 68%, respectively. Collagen gel contraction was reduced by 36% (P < 0.001), 27% (P = 0.0003), and 33% (P = 0.0019) following treatment with fasudil (25 μM), Y27632 (10 μM), and H1152 (10μM). ROCK activity induced by TGFβ rose 4.74 ± 1.9 times over control at 4 hours (P = 0.0004) and 2.4 ± 0.47-fold (P = 0.0016) in sclera after IOP elevation. Proliferation of scleral fibroblasts after chronic IOP elevation was reduced 77% by Y27632 (P = 0.001) and 84% by fasudil (P = 0.0049).ROCK inhibitors reduce TGFβ-induced myofibroblast transdifferentiation and glaucoma-induced scleral cell proliferation.These findings suggest altered fibroblast activity promoted by ROCK inhibitors could modify scleral biomechanics and be relevant to glaucoma treatment.
Fasudil
Rho kinase inhibitor
Transdifferentiation
Myofibroblast
Sclera
Proliferative Vitreoretinopathy
ROCK1
Cite
Citations (44)
There is growing evidence that the Rho/Rho-kinase (ROCK) signaling pathway is upregulated in tumors and plays a key role in cancer invasion and proliferation. The aim of this study was to explore the anti-tumor effects of Rho/ROCK inhibitor, fasudil, including the possible mechanisms involved in the suppression of the glioblastoma (GBM) cell line progression in vitro and in vivo. After T98G and U251 cells were treated with various concentrations of fasudil, Y27632, and ROCK siRNA, the effects of ROCK inhibitors on migration, invasion, invasion-related gene expressions, proliferation, and apoptosis of cultured tumor cells were examined. The results indicated that fasudil significantly inhibited not only proliferation, migration, and invasiveness (P < 0.05) but also the mRNA and protein expressions of matrix metalloproteinase-2 (MMP-2) in a dose-dependent manner. Moreover, fasudil treatment resulted in a dose-dependent increase of apoptosis in T98G and U251. The intracranial xenograft models were established. The cryosection of the tumor and the survival time of mice in each group indicated that fasudil could inhibit glioma invasion and growth in vivo. Based on the results, fasudil suppresses the progression of GBM in vitro and in vivo by inhibiting ROCK. This could be linked to the decreased MMP-2 expression and the induction of apoptosis in tumor cells. The Rho/ROCK signaling pathway may prove to be a promising target in anti-tumor therapy. Fasudil may be an attractive anti-tumor drug candidate for the treatment of GBM.
Fasudil
Rho kinase inhibitor
U87
MMP9
Tumor progression
Cite
Citations (73)
Fasudil
Rho kinase inhibitor
Cite
Citations (111)
Objective:To investigate the effect of Rho-ROK inhibitors on axon regeneration of mouse cortical neurons after hypoxia Methods: Mouse cortical neurons were cultured and treated with hypoxia.The inhibitors of Rho kinase,Y-27632,Fasudil and Exoenzyme C3a,were applied to the cultures before and after hypoxia.The axon regeneration was determined by immunostaining of F-actin.The neuronal viability was measured by MTT method.Results: In control normal neuronal cultures,axons and dendrites formed the network,and the actin cytoskeleton was distributed in the periphery of neurons and formed the actin bands.Stress fibers were rarely observed in the endochylema.However,after exposure to hypoxia,actin bands became blurred and the neuronal processes were retracted.The cells were swollen and became apoptotic.The cellular changes induced by hypoxia could be reversed by Y-27632 and fasudil whereas Exoenzyme C3a showed little effects.Y-27632 and fasudil also could improve the viability of neurons after hypoxia.Conclusion: Hypoxia induces the recombination of cytoskeleton and collapse of growth cones in parallel with apoptosis of cortical neurons.Suppression of Rho kinase activity with different Rho-ROK inhibitors could inhibit this process.Y-27632 and fasudil show significant effects but exoenzyme C3a displays only a marginal effect.
Fasudil
Rho kinase inhibitor
Hypoxia
Cofilin
Cite
Citations (0)
RhoA/Rho kinase 小径可以参予的背景组织缺氧和 monocrotaline 的致病导致了肺的高血压。这研究测试了 RhoA/Rho kinase 小径是否涉及高流动的致病在 rats.Methods 男性 Wistar 老鼠(4 个星期) 的导致的肺的高血压随机被划分成 4 个分流组， 4 个对待的组和对待的组经历了左普通颈动脉 artery/external 颈部的静脉分流操作。控制组经历了假冒的操作。对待的组接受了 fasudil 治疗，其它收到了一样的剂量盐。在星期 1 研究， 2， 4 和 8，恰好室的收缩压力被测量，血气体被分析计算 Qp/Qs。到加中隔和在中等大小的肺的动脉的中间的墙厚度的吝啬的百分比的左室的正确的室的重量比率被获得。在肺的动脉的 RhoA 活动用 Rho 活动试金试剂被检测。 Rho kinase 活动被 MYPT1 phosphorylation 的程度与西方的 blot.Proliferating 房间确定用增殖的房间被评估原子抗原 immunohistological staining.Results 颈动脉 artery/jugular 静脉分流导致了高肺的血流动，两一尖锐并且恰好室的收缩压力的长期的举起，重要的中间的墙变厚由光滑的肌肉房间增长，恰好室的肥大和 RhoA 和 Rh 的增加的激活描绘了 Fasudil 治疗降低了肺的动脉收缩压力，压制的肺的动脉光滑的肌肉房间增长，中间的墙和 Rho kinase 活动然而并非 Rho activity.Conclusions 的重要抑制变厚和禁止的恰好室的肥大激活的稀释肺的动脉 RhoA/Rho kinase 小径与尖锐肺的血管缩小和高流动改变的长期的肺的动脉被联系导致了肺的高血压。Fasudil 处理能由禁止 Rho kinase 活动改进肺的高血压。
Fasudil
Rho kinase inhibitor
ROCK2
Cite
Citations (0)
Cardiac allograft vasculopathy (CAV) continues to be a major cause of late graft failure after cardiac transplantation. We have demonstrated that Rho-kinase, an effector of the small GTPase Rho, plays an important role in the pathogenesis of arteriosclerosis. In this study, we examined whether the Rho-kinase–mediated pathway is also involved in the pathogenesis of CAV using a specific Rho-kinase inhibitor and a dominant-negative Rho-kinase. Hearts from AKR mice were heterotopically transplanted to C3H/He (allograft) or AKR mice (isograft), and the effects of long-term oral treatment with fasudil, which is metabolized to a specific Rho-kinase inhibitor hydroxyfasudil, on CAV were examined at 2 and 4 weeks after the transplantation. Coronary remodeling in the allografts characterized by intimal thickening and perivascular fibrosis was dose-dependently suppressed in the fasudil group compared with the control group (P<0.01, n=9 to 10). The inhibitory effects of hydroxyfasudil were mimicked by in vivo gene tr...
Fasudil
Rho kinase inhibitor
Pathogenesis
Isograft
Arteriosclerosis
Ex vivo
Cite
Citations (0)
Rho-associated kinase (ROCK) is a key regulatory protein involved in inflammatory secretion in microglia in the central nervous system. Our previous studies showed that ROCK inhibition enhances phagocytic activity in microglia through the extracellular signal-regulated kinase (ERK) signaling pathway, but its effect on microglial migration was unknown. Therefore, in this study, we investigated the effects of the ROCK inhibitors Y27632 and fasudil on the migratory activity of primary cultured microglia isolated from the spinal cord, and we examined the underlying mechanisms. The microglia were treated with Y27632, fasudil and/or the ERK inhibitor U0126. Cellular morphology was observed by immunofluorescence. Transwell chambers were used to assess cell migration. ERK levels were measured by in-cell western blot assay. Y27632 and fasudil increased microglial migration, and the microglia were irregularly shaped and had many small processes. These inhibitors also upregulated the levels of phosphorylated ERK protein. The ERK inhibitor U0126 suppressed these effects of Y27632 and fasudil. These findings suggest that the ROCK inhibitors Y27632 and fasudil promote microglial migration in the spinal cord through the ERK signaling pathway.
Fasudil
Rho kinase inhibitor
Cite
Citations (21)