Dexmedetomidine alleviates lung tissue injury of rat models with ventilator-associated lung injury / 基础医学与临床

Objective To investigate the effect of dexmedetom idine(DEX)on lung tissue and Ras homolog gene family member A(RhoA)/Rho kinase 1(ROCK1)signaling pathway in lung tissue of rats with ventilator-induced lung injury(VILI).Methods A VILI rat model was established and separated into control group,model group(VILI group),dexmedetomidine low and high dose groups(DEX-L,DEX-H group),and high dose dexmedetomi-dine+lysophosphatidic acid(LPA)group(DEX-H+LPA group).Determination of wet/dry mass ratio of rat lung tissue(W/D);HE staining microscopy was applied to observe morphology of lung tissue;ELISA kit was applied to detect the level of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in bronchoalveolar lavage fluid(BALF);TUNEL staining method was applied to detect lung epithelial cell death;Immunoblotting was applied to detect the expression levels of apoptosis-related proteins,and RhoA,ROCK1 pro-teins.Results DEX could reduce lung injury,lung injury score,W/D,apoptosis rate,levels of TNF-α,IL-1β,IL-6,and expression of Bax,cleaved caspase-3,RhoA,ROCK,α-SMA in VILI rats(P＜0.05),while increased the expression of Bcl-2(P＜0.05);LPA could aggravate lung injury and increase lung injury score,W/D,apopto-sis rate,level of TNF-α,IL-1β,IL-6 and expressions of Bax,cleaved caspase-3,RhoA,ROCK and α-SMA(P＜0.05);Bcl-2 expression level was decreased(P＜0.05).Conclusions Dexmedetomidine may protect rats with ventilator-induced lung injury by the inhibition of RhoA/ROCK1 signaling pathway.

Role of ripasudil as an adjunct treatment in the management of pseudoexfoliative glaucoma

Purpose: Ripasudil is a class of drug which alters the trabecular meshwork to increase the aqueous outflow and has been shown to be effective in pseudoexfoliative glaucoma (PXF G). This study aimed at assessing the efficacy and safety profile of ripasudil as an adjunct treatment in patients with PXF G at maximal tolerated antiglaucoma medications. Methods: In this prospective, interventional study, 40 patients with PXF G were enrolled between May 2021 and Jan 2022. Ripasudil 0.4% was started as an adjunctive drug to the ongoing antiglaucoma medications. On follow?up visits at 1, 3, and 6 months, the visual acuity, intraocular pressure (IOP), anterior segment, and fundus findings were evaluated. The premedication and postmedication IOP values were compared by paired t?test, and a P?value <0.05 was considered statistically significant. Results: Average age at recruitment was 60.02 ± 8.74 years. Baseline premedication IOP was 25.375 ± 3.276 mmHg. IOP reduction at 6 months was found to be statistically significant in all patients, with the maximal response being 24.13%. Also, 87.5% (35/40) of patients reached target IOP or even lower IOP at the end of study. There was no statistically significant association between the PXF grade and IOP. However, the grade of inferior iridocorneal angle pigmentation was found to be higher in eyes with elevated IOP (P < 0.05). Only three patients developed conjunctival hyperemia as an adverse reaction, which was mild and transient. Conclusion: Ripasudil showed additional IOP?lowering effect with other antiglaucoma medications and exhibited no significant side effects

Netarsudil monotherapy as the initial treatment for open-angle glaucoma and ocular hypertension in Indian patients: A real-world evaluation of efficacy and safety

Purpose: Glaucoma is the second leading cause of blindness worldwide, affecting more than 64 million people aged 40–80. The best way to manage primary open?angle glaucoma (POAG) is by lowering the intraocular pressure (IOP). Netarsudil is a Rho kinase inhibitor, the only class of antiglaucoma medications that reorganizes the extracellular matrix to improve the aqueous outflow through the trabecular pathway. Methods: An open?label, real?world, multicentric, observation?based 3?month study was performed for assessing the safety and ocular hypotensive efficacy of netarsudil ophthalmic solution (0.02% w/v) in patients with elevated IOP. Patients were given netarsudil ophthalmic solution (0.02% w/v) as a first?line therapy. Diurnal IOP measurements, best?corrected visual acuity, and adverse event assessments were recorded at each of the five visits (Day?1: screening day and first dosing day; subsequent observations were taken at 2 weeks, 4 weeks, 6 weeks, and 3 months). Results: Four hundred and sixty?nine patients from 39 centers throughout India completed the study. The mean IOP at baseline of the affected eyes was 24.84 ± 6.39 mmHg (mean ± standard deviation). After the first dose, the IOP was measured after 2, 4, and 6 weeks, with the final measurement taken at 3 months. The percentage reduction in IOP in glaucoma patients after 3 months of once?daily netarsudil 0.02% w/v solution use was 33.34%. The adverse effects experienced by patients were not severe in the majority of cases. Some adverse effects observed were redness, irritation, itching, and others, but only a small number of patients experienced severe reactions, as reported in a decreasing order: redness > irritation > watering > itching > stinging > blurring. Conclusion: We found that netarsudil 0.02% w/v solution monotherapy when used as the first?line treatment in primary open?angle glaucoma and ocular hypertension was both safe and effective.

Inhibidores de la Rho quinasa y su rol en la disfunción endotelial corneal / Rho Kinase Inhibitors and their Role in Corneal Endothelial Dysfunction

El endotelio corneal es su capa más interna y, a pesar de ser una monocapa de células, es capaz de preservar la transparencia del tejido con dos funciones fundamentales: de barrera y de bomba endotelial sodio-potasio (Na-K). Las células endoteliales tienen muy poca capacidad de regeneración, por lo que cualquier lesión endotelial es compensada por la expansión y migración de las células residuales adyacentes. La disfunción endotelial corneal se caracteriza por un edema de la córnea que puede llevar hasta el transplante de este tejido. Nuevas terapias farmacológicas con inhibidores de Rho-Kinasa y terapias basadas en ingeniería tisular se han propuesto recientemente. Se realizó una búsqueda automatizada sobre los principales avances en estas terapias utilizando la plataforma Infomed, específicamente la Biblioteca Virtual de Salud. La información se resumió en el informe final. Concluimos que existe un progreso significativo en el entendimiento de la patogénesis, y en el desarrollo de los nuevos tratamientos(AU)

The corneal endothelium is its innermost layer and, despite being a monolayer of cells, is able to preserve tissue transparency with two fundamental functions: barrier and endothelial sodium-potassium (Na-K) pump. Endothelial cells have very little regenerative capacity, so any endothelial injury is compensated by the expansion and migration of adjacent residual cells. Corneal endothelial dysfunction is characterized by corneal edema that can lead to corneal tissue transplantation. New pharmacologic therapies with Rho kinase inhibitors and tissue engineering-based therapies have recently been proposed. An automated search on the main advances in these therapies was performed using the Infomed platform, specifically the Virtual Health Library. The information was summarized in the final report. We conclude that there is significant progress in the understanding of pathogenesis, and in the development of new treatments(AU)

Evaluation of the Rho-kinase gene expression and polymorphisms in adult patients with acute appendicitis: a differential impact of gender

SUMMARY OBJECTIVE: Acute appendicitis represents one of the most common causes of acute intra-abdominal emergencies worldwide. In this case-control study, we aimed to investigate associations of Rho-kinase gene expression and polymorphisms with acute appendicitis in a Turkish population. We also aimed to study the effects of gender on these parameters. METHODS: A total of 93 unrelated patients with acute appendicitis and 93 healthy controls in the Department of Emergency Medicine, Erciyes University, between June 2019 and June 2021 were included in this study. Genomic DNA was isolated from peripheral leukocytes, and the LightCycler 480 II real-time polymerase chain reaction was utilized to detect Rho-kinase1 gene rs35996865 and Rho-kinase2 gene rs2230774 (Thr431Asn) polymorphisms. Quantitative real-time polymerase chain reaction was applied to determine Rho-kinase1 and Rho-kinase2 gene expressions. RESULTS: There was a marked increase in Rho-kinase1, but not in Rho-kinase2, mRNA expression, and this increase was evident only in male patients (p=0.0008). No significant differences were found in allele and genotype frequencies for Rho-kinase1 gene rs35996865 and Rho-kinase2 gene rs2230774 polymorphisms between the patients with acute appendicitis and the control group. CONCLUSIONS: Our data imply that Rho-kinase1 (rs35996865) and Rho-kinase2 (rs2230774) gene variants are not risk factors for the development of acute appendicitis in the Turkish population. However, increased mRNA expression of the Rho-kinase1 gene in males indicated that Rho-kinase1 is involved in the pathogenesis of acute appendicitis in a gender-specific way.

Safety, efficacy, and patient selection of ripasudil in patients with uncontrolled glaucoma with maximum conventional medical therapy

Purpose: Ripasudil hydrochloride hydrate (0.4%) is the first Rho?associated protein kinase (ROCK) inhibitor eye drop that lowers intraocular pressure (IOP) by increasing conventional aqueous outflow through the trabecular meshwork and Schlemm’s canal. We aimed to evaluate the safety and efficacy of ripasudil in patients using the maximum topical anti?glaucoma medications and with uncontrolled IOP. Methods: In our prospective interventional study, we enrolled 27 eligible and consenting patients (46 eyes) who presented to us between January 2021 and June 2021. Ripasudil 0.4% was added as adjunctive therapy to the ongoing glaucoma treatment. On follow?up visits at 7 days, 15 days, 1 month, 2 months, and 3 months, the visual acuity, IOP with applanation tonometer, anterior segment, and fundus were evaluated. The IOP before and after the use of ripasudil eye drops was compared by paired t?test. Results: Among the 27 patients, 18 were males and 9 were females. A statistically significant reduction in IOP was noted at all time durations (P < 0.00001) with the maximum reduction at 3 months with all patients achieving their target IOP. No patient developed any side effects necessitating the omission of ripasudil. The most common adverse event noted was conjunctival hyperemia (22 patients), which was mild and transient. Conclusion: Ripasudil showed additional IOP?lowering effect with other antiglaucoma medications and exhibited no significant side effects.

Reticular epithelial corneal edema as a novel side-effect of Rho Kinase Inhibitors: An Indian scenario

Purpose: To describe clinical course, characteristics, and outcome of reticular epithelial corneal edema (RECE) occurring as a not?so?infrequent adverse effect of a novel drug, Rho?kinase inhibitors (ROCK?I)? netarsudil (0.02%) and ripasudil (0.4%). Methods: This was a retrospective observational non?randomized study. In this study, 12 eyes of 11 patients presenting at a tertiary eye care center between April 2021 and September 2021 were included. All 12 eyes developed a distinctive honeycomb pattern of RECE after starting topical ROCK?I. All patients were subjected to detailed ophthalmic examinations. Results: Eight patients were started on netarsudil (0.02%) and three on ripasudil (0.4%). Five eyes had a prior history of corneal edema. The remaining seven had the presence of ocular comorbidities predisposing to corneal edema. The average time for RECE occurrence was 25 days for netarsudil and 82 days for ripasudil. Visual acuity decreased in two eyes, remained unaffected in four eyes, and could not be quantified in four eyes due to preexisting profound visual impairment. Five eyes had symptoms of ocular surface discomfort associated with bullae. Symptoms and bullae resolved in all eyes in whom ROCK?I was stopped. The average time to resolution of RECE was 10 days for netarsudil and 25 days for ripasudil. Conclusion: RECE after ROCK?I occurs with the use of both netarsudil and ripasudil, although the characteristics differ. The presence of corneal edema and endothelial decompensation seem to be a risk factor, and cautious use is warranted in these patients. Four clinical stages of RECE are described. ROCK?I act as a double?edged sword in patients with endothelial decompensation. Large?scale studies are required to know the exact incidence, pathophysiology, and long?term consequences of the aforementioned side?effect.

K-115 enhances autophagic activity and attenuates TGF-β1-induced activation of human Tenon's fibroblasts / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the influence of K-115 on the proliferation and migration of human Tenon's fibroblasts(HTFs)and to access the possible mechanism. Furthermore, to provide new ideas for anti-scar treatment after glaucoma surgery.METHODS: The Tenon capsule tissues were collected from patients who underwent glaucoma surgery in Hebei General Hospital from September 2018 to September 2019. Primary culture of HTFs was performed by tissue block method. The transforming growth factor-β1(TGF-β1)was used to induce HTFs activation that can mimic glaucoma filtration surgery. The cells were treated with K-115 and divided into 4 groups: the control group was treated with dimethyl sulfoxide(DMSO); TGF-β1 group was treated with 10μg/L TGF-β1 for 24h; TGF-β1 +5 K-115 group was pretreated with 5μmol/L K-115 for 2h and then treated with 10μg/L TGF-β1 for 24h; TGF-β1+10 K-115 group was pretreated with 10μmol/L K-115 for 2h and then 10μg/L TGF-β1 was added for 24h. Cell proliferation was observed by cell proliferation experiment. The migration ability of cells was detected by scratch test. The formation of autophagosomes was observed by transmission electron microscopy. Apoptosis was visualized by Hoechst 33342/PI staining.RESULTS: Cell proliferation experiment revealed that K-115 could inhibit the proliferation of HTFs induced by TGF-β1. Scratch test suggested that K-115 could inhibit the migration of HTFs induced by TGF-β1. Transmission electron microscope results showed that K-115 could enhance autophagy of HTFs induced by TGF-β1. Hoechst 33342/PI staining suggested that K-115 did not induce apoptosis.CONCLUSIONS: K-115 may regulate the proliferation and migration of HTFs induced by TGF-β1 by increasing autophagy rather than inducing apoptosis.

Effect and mechanism of Rho kinase inhibitor on intestinal injury in septic rats / 中华危重病急救医学

Objective:To explore the effect of Rho kinase inhibitor on intestinal injury in septic rats and its possible mechanism.Methods:Thirty-two male Sprague-Dawley (SD) rats were randomly divided into sham operation group (Sham group), Rho kinase inhibitor Y-27632 control group (Y+Sham group), sepsis model group [cecal ligation and puncture (CLP) group] and Y-27632 pretreatment group (Y+CLP group), with 8 rats in each group. Rat sepsis model was reproduced by CLP. The rats in the Sham group and Y+Sham group were only separated and moved the cecum without ligation and perforation. The rats in the Y+Sham group and Y+CLP group were pretreated with intraperitoneal injection of Y-27632 solution 5 mg/kg 15 minutes before operation; the rats in the Sham group and CLP group were intraperitoneally injected with the same amount of phosphate buffered saline (PBS). Twenty-four hours after operation, the heart blood was collected and the serum diamine oxidase (DAO) content was determined by enzyme-linked immunosorbent assay (ELISA). Then the small intestine tissue was collected, the pathological changes of the intestinal tissue were observed under the light microscope after hematoxylin-eosin (HE) staining, and Chiu's score was performed. The positive expressions of Rho-related coiled-coil kinase 1 (ROCK1) and nuclear factor-κB (NF-κB) in intestinal tissue were detected by immunohistochemistry. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in intestinal tissue homogenate.Results:The intestinal tissue structure of the Sham group and Y+Sham group was intact and the mucosa was arranged neatly. Compared with the Sham group, the intestinal mucosa of the CLP group was arranged disorderly, with a large number of inflammatory cells infiltration, and the Chiu's score was significantly increased (3.83±0.27 vs. 0.12±0.11, P < 0.05), indicating that those rats suffered from septic intestinal injury. Compared with the CLP group, the degree of necrosis of intestinal epithelial cells in the Y+CLP group was reduced, a small amount of inflammatory cells infiltration was seen, and the Chiu's score was significantly decreased (2.85±0.21 vs. 3.83±0.27, P < 0.05), indicating that Y-27632 pretreatment could alleviate intestinal injury in septic rats. Compared with the Sham group, the positive expressions of intestinal tissue ROCK1 and NF-κB, the contents of serum DAO and intestinal homogenate TNF-α in the CLP group were significantly increased [ROCK1 expression ( A value): 0.19 (0.18, 0.22) vs. 0.10 (0.09, 0.11), NF-κB expression ( A value): 0.40±0.02 vs. 0.15±0.01, DAO (ng/L): 287.81±23.31 vs. 144.92±17.72, TNF-α (ng/L): 101.08±5.62 vs. 74.81±5.56, all P < 0.05], the level of intestinal homogenate IL-10 was significantly decreased (μg/L: 55.16±5.20 vs. 95.95±7.53, P < 0.05). Compared with the CLP group, the positive expressions of intestinal tissue ROCK1, NF-κB, the contents of serum DAO and intestinal homogenate TNF-α in the Y+CLP group were significantly decreased [ROCK1 expression ( A value): 0.15 (0.13, 0.18) vs. 0.19 (0.18, 0.22), NF-κB expression ( A value): 0.28±0.01 vs. 0.40±0.02, DAO (ng/L): 243.34±19.76 vs. 287.81±23.31, TNF-α (ng/L): 90.41±8.79 vs. 101.08±5.62, all P < 0.05], while the level of intestinal homogenate IL-10 was significantly increased (μg/L: 66.15±5.74 vs. 55.16±5.20, P < 0.05), indicating that the protective effect of Y-27632 pretreatment on sepsis intestinal injury rats might be related to the regulation of RhoA/ROCK1/NF-κB signaling pathway. Conclusion:Rho kinase inhibitors can reduce intestinal injury in septic rats, and the mechanism may be related to inhibiting RhoA/ROCK1/NF-κB signaling pathway and reducing intestinal inflammation in septic rats.

Evaluation of the preventive effect of DL0805-2 against monocrotaline induced rat pulmonary arterial hypertension / 药学学报

In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury. This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups (1, 3, and 10 mg·kg-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues. DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.

The protective mechanisms of hippocampal neurons on RhoA/ROCK2 pathway mediated by Rho kinase inhibitors in vascular dementia rats / 中风与神经疾病杂志

@#Objective　To investigate the protective mechanism of Rho protein kinase 2 (ROCK2) inhibitor constructed based on adeno-associated virus vector on vascular dementia (VaD) rat hippocampal neurons.Methods　Forty SD rats were randomly divided into sham operation group,model group,ROCK2 interference group (shROCK2),negative control group.The VaD model was prepared by permanent ligation of bilateral common carotid arteries.AAV9-ROCK2-shRNA was injected into the hippocampus of the rat by stereotaxic technique,the negative control group was injected with the same amount of empty vector adeno-associated virus.After 4 weeks,morris water maze was used to test the learning and memory abilities of rats.RT-PCR and Western blot were used to detect the mRNA and protein expression levels of RhoA,ROCK2,MLC,MLCP.Results　Four weeks after operation.The mRNA and protein expressions of the above 4 indicators in hippocampus increased (P<0.01),the synaptic vesicles disappeared,and the structure of hippocampus was chaotic in the model group.Compared with the model and control group,the learning and memory ability of rats in ROCK2 interference group was significantly improved,the mRNA and protein of the above 4 indicators were significantly decreased (P<0.05),and the pre-synaptic vesicles were normal and the organelles were more complete.Conclusion　Rho protein kinase 2 inhibitors can reduce the expression of downstream related proteins by inhibiting RhoA/ROCK2 signaling pathway,then promote neuronal axon regeneration and alleviate cognitive dysfunction of VaD rats.

Meta-analysis of Rho kinase inhibitor Ripasudil in the treatment of glaucoma / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To evaluate the efficacy and safety of ripasudil in the treatment of glaucoma in recent years by Meta-analysis.<p>METHODS:According to the Cochrane systematic review method, we searched three databases(Pubmed, Web of Knowledge and CNKI). The change of intraocular pressure was used to assess the effect of ripasudil, WMD with 95% confidence intervals(<i>CI</i>)were used to report the outcome. Subgroup analysis was performed according to different types of glaucoma.<p>RESULTS:A total of 9 trials all in English, totally 3446 patients, Meta-analysis showed that compared with before-treated groups, after-treated groups have significant lower IOP(Test of WMD= -2.96 95% <i>CI</i>: -3.48 to -2.45, <i>P</i><0.001), which indicates that ripasudil has great efficacy of curing glaucoma.<p>CONCLUSION:Ripasudil have significant efficacy in reducing IOP, which can be used as a treatment for glaucoma.

Netarsudil: a novel intra-ocular pressure lowering agent

Optic disc health is an important indicator of visual functions. The first line of management to prevent/halt the damage to optic disc is to control responsible pathological condition, if identified. In absence of identifiable cause, the most validated approach is lowering of intra-ocular pressure (IOP). Individually, as well as combinations of currently available drugs are not fully effective in all patients of glaucoma in achieving desired IOP control. Hence, there is a need of newer alternatives which address this unmet need. Recently, a newer IOP lowering agent with a novel mechanism of action, netarsudil, has been approved by United States Food and Drug Administration (US-FDA) in December 2017. Netarsudil acts by inhibiting Rho-associated protein kinase resulting in lowering of overall tone of the contractible cells in trabecular meshwork thereby improving drainage of aqueous humor outflow and lowering of IOP. Though in its early days, this drug gives an armamentarium to ophthalmologists and physicians to control IOP in patients of open-angle glaucoma and ocular hypertension.

Mitochondrial oxidative damage and apoptosis induced by high glucose through Rho kinase signal pathway in renal tubular epithelial cells

Objective:To investigate the role of oxidative stress in human renal tubular epithelial cells (HK-2) induced by high glucose and the underlying signal pathway in vitro.Methods:MYPT1, pro-caspase-3, PGC-1α, and Drp1 protein expressions were measured by Western blot. MnSOD2, Drp1 and PGC-1α mRNA expressions were detected by real time PCR.Results:Results showed that high glucose significantly up-regulated the protein expressions of MYPT1, pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells; while Rho kinase inhibitor fasudil and ROCK1 siRNA inhibited protein expressions of pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells induced by high glucose. Importantly, fasudil and ROCK1 siRNA markedly inhibited the expressions of mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α in HK-2 cells induced by high glucose.Conclusions:Our findings suggest that Rho kinase signal pathway is involved in mitochondrial oxidative damage and apoptosis in high glucose-induced renal tubular epithelial cells by regulating mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α. Targeting Rho kinase signal pathway might be a potential strategy for the treatment of diabetic nephropathy.

Mitochondrial oxidative damage and apoptosis induced by high glucose through Rho kinase signal pathway in renal tubular epithelial cells

Objective: To investigate the role of oxidative stress in human renal tubular epithelial cells (HK-2) induced by high glucose and the underlying signal pathway in vitro. Methods: MYPT1, pro-caspase-3, PGC-1α, and Drp1 protein expressions were measured by Western blot. MnSOD2, Drp1 and PGC-1α mRNA expressions were detected by real time PCR. Results: Results showed that high glucose significantly up-regulated the protein expressions of MYPT1, pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells; while Rho kinase inhibitor fasudil and ROCK1 siRNA inhibited protein expressions of pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells induced by high glucose. Importantly, fasudil and ROCK1 siRNA markedly inhibited the expressions of mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α in HK-2 cells induced by high glucose. Conclusions: Our findings suggest that Rho kinase signal pathway is involved in mitochondrial oxidative damage and apoptosis in high glucose-induced renal tubular epithelial cells by regulating mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α. Targeting Rho kinase signal pathway might be a potential strategy for the treatment of diabetic nephropathy. http://www.apjtm.org/article.asp?issn=1995-7645;year=2018;volume=11;issue=6;spage=399;epage=404;aulast=Li;type=2.

Therapeutic potential of Rho kinase in diabetic retinopathy / 眼科新进展

Rho-associated kinase is originally identified as an effector protein of the G protein Rho,which involves in various diseases,particularly in cancer and cardiovascular disease.Rho kinase inhibitors have already been applied clinically for cerebral vasospasm and glaucoma.Diabetic retinopathy (DR) is a common complication of diabetes which is the leading cause of visual loss.While anti-VEGF therapy has recently been widely used for DR patients due to its efficacy,but great attention has been drawn to the related risk and complications.The importance of Rho kinase in pathological vitreoretinal conditions has also been elucidated and is attracting attention as a potential therapeutic target.Rho kinase is involved in anglogenesis and hyperpermeability and also in the pathogenesis of various pathologies such as inflammation and neural degeneration,which has been expected that Rho kinase inhibitors will become a new molecular target drug for DR.This review summarizes the mechanism of Rho kinase action and its application in the treatment of DR.

Mechanism of coronary artery constriction induced by 5-HT / 中国病理生理杂志

AIM:To investigate the possible mechanism of coronary artery contraction induced by 5-hydroxytryptamine(5-HT).METHODS:Isolated coronary artery rings were obtained from male Wistar rats,and the vas-cular tension meter was used to determine the tension of the coronary artery rings.The effects of inhibitors of different sig-naling pathway on vascular contraction tension induced by 5-HT were observed.RESULTS:Firstly,we found that 5-HT2A receptor antagonist sarpogrelate(1 μmol/L)completely eliminated the coronary artery contraction induced by 5-HT.Phos-pholipase Cβ(PLCβ)inhibitor U73122(10 μmol/L and 50 μmol/L), Rho-related protein kinase inhibitor Y-27632(3 μmol/L and 10 μmol/L)and protein kinase C δ subunit(PKCδ)inhibitor rottlerin(3 μmol/L and 10 μmol/L)signifi-cantly inhibited the contraction of coronary artery ring caused by 5-HT(P<0.05).In addition, compared with the un-treated group,vascular contraction tension induced by 5-HT was also decreased significantly by L-type calcium channel (Cav1.2)blocker nifedipine(1 μmol/L), store-operated Ca2+entry(SOCE)inhibitor SKF96365(10 μmol/L and 30 μmol/L)and 2-aminoethoxydiphenyl borate(2-APB,50 μmol/L and 100 μmol/L)(P<0.05).At the same time,5-HT also induced vasoconstriction after treated with nifedipine(1 μmol/L)Kerbs-Henseleit(K-H)liquid without calcium (P<0.05).CONCLUSION:5-HT activates 5-HT2Areceptor induced coronary artery contraction,possibly related to the PKC/Rho kinase signaling pathway and calcium regulation.

Rho-kinase Inhibitor Ameliorates Lipopolysaccharide-induced Kidney Injury by Inhibiting Toll-like Receptor 4 and Nuclear Factor-κB Signaling Pathway / 中国医科大学学报

Objective To explore whether Rho kinase inhibitor protects endotoxemia mice from kidney injury,and to investigate the mechanism underlying this effect. Methods Adult male C57BL/6 mice were randomly divided into three groups (n = 8 for each group): control,lipopolysaccharide (LPS),and LPS+ Y-27632 (Rho kinase inhibitor). For induction of acute kidney injury,mice were administered 30 mg/kg LPS intraperitoneally. Y-27632 (10 mg/kg body weight) was injected intraperitoneally 18 h and 1 h before injection of LPS,and an equal volume of sterile saline was administered at the corresponding time point in each group. The mice were killed 8 h after LPS administration. Blood samples and kidney tissues were taken and preserved for subsequent analysis. Results Pretreatment with Y-27632 significantly attenuated LPS-induced kidney injury;pretreatment with Y-27632 markedly reduced renal expression of inflammatory cytokines (TNF-α and IL-1β) in endotoxemia mouse,and also significantly inhibited LPS-induced caspase-3 expression in the kidney; and Y-27632 pretreatment dramatically reduced TLR4 protein expression and NF-κBp65 phosphorylation in kidney tissues of endotoxemia mouse. Conclusion Rho kinase inhibitor may inhibit TLR4 and NF-κB signaling pathway to reduce the inflammatory response in the kidneys of endotoxemia mice and alleviate acute renal injury induced by LPS.

Effect and significance of tetrandrine on expression of Rho/Rho kinase in fetal lung of congenital diaphragmatic hernia rats / 重庆医学

Objective To investigate the effect and significance of the traditional Chinese medicine tetrandrine(TET) prenatal intervention on the expression of RhoA protein and Rho kinase ROCK1 in the fetal lung of congenital diaphragmatic hernia (CDH) rat model.Methods SD female rats with 9.5 d of gestation were randomly divided into the control group,CDH group and TET group.The CDH group and TET intervention group were administered with nitrofen by gavage for establishing CDH model.The TET intervention was given on 16.5 d of gestation.The fetal rat lungs were taken by cesarean section on 21.5 d of gestation and the lung weight/body ratio(Lw/Bw) was measured.The lung development and small pulmonary arterial morphologic changes in HE staining in all groups were observed with microscopy.The protein expression of RhoA and Rho kinase ROCK1were respectively examined by immunohistochemistry and Western blot.Results In the CDH group,the lungs had obvious maldevelopment and the fetal lung development in the TET group was close to that in the control group.The lung development indicators of Lw/Bw,PAA％,and lung vascular remodeling indicators of lumen area and vascular total area ratio(LA％) in the CDH group were significantly lower than those in the control group(2.11±0.36 vs.4.24±0.31;33.60±3.12 vs.58.81 ±2.92;38.58±2.15 vs.61.20±3.23,P＜0.05),the indicators of Lw/Bw.PAA％ and LA％ after TET intervention were significantly improved compared with the CDH group(3.61±0.24 vs.2.11±0.36;42.46±3.68 vs.33.60±3.12;56.07±3.32 vs.38.58±2.15,all P ＜0.05);the ratio of small pulmonary artery wall thickness to vascular external diameter (WT％) and the medium thickness percentage (MT％) in CDH group were significantly higher than those in the control group(26.64±2.41 vs.13.50±1.45 and 25.98±2.79 vs.16.47±2.07,P＜0.05),WT％ and MT％ in the TET group were obviously lower than those in the CDH group (16.02±2.35 vs.26.64± 2.41 and 17.96 ± 1.95 vs.25.98 ± 2.79,P＜0.05).The immunohistochemistry and Western blot detection indicated that the expressions of RhoA and ROCK1 from low to high were the control group ＜TET group ＜ CDH group.Conclusion Pulmonary hypoplasia and lung vascular remodeling exist in fetal rats with CDH and Rho/Rho kinase signaling pathway may be involved in the process.Prenatally giving TET may play the lung protective effect by regulating the Rho/Rho signal pathway.

Rho kinase signal pathway and inflammatory bowel disease / 国际儿科学杂志

Rho/Rho-kinase signaling pathway is a ubiquitous one in vivo and plays a role of "molecular switch" by regulating the polymeric state of intracellular actin cytoskeleton and participates in the regulation of various cellular functions.As research continues,some of them have provided evidence of Rho kinase signal pathway function at the occurrence and development of inflammatory bowel disease.This review aims to summarize the relationship between Rho kinase signal pathway and inflammatory bowel disease.