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OBJECTIVE: Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage. Notably, the knockout of transient receptor potential channel 6 (TRPC6) has shown promise in alleviating renal fibrosis. However, the regulatory impact of TRPC6 on renal fibrosis remains unclear. METHODS: In vivo, TRPC6 knockout (TRPC6-/-) mice and age-matched 129 SvEv (WT) mice underwent unilateral renal ischemia-reperfusion (uIR) injury surgery on the left renal pedicle or sham operation. Kidneys and serum were collected on days 7, 14, 21, and 28 after euthanasia. In vitro, primary tubular epithelial cells (PTECs) were isolated from TRPC6-/- and WT mice, followed by treatment with transforming growth factor ß1 (TGFß1) for 72 h. The anti-fibrotic effect of TRPC6-/- and the underlying mechanisms were assessed through hematoxylin-eosin staining, Masson staining, immunostaining, qRT-PCR, and Western blotting. RESULTS: Increased TRPC6 expression was observed in uIR mice and PTECs treated with TGFß1. TRPC6-/- alleviated renal fibrosis by reducing the expression of fibrotic markers (Col-1, α-SMA, and vimentin), as well as decreasing the apoptosis and inflammation of PTECs during fibrotic progression both in vivo and in vitro. Additionally, we found that the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3 beta (GSK3ß) signaling pathway, a pivotal player in renal fibrosis, was down-regulated following TRPC6 deletion. CONCLUSION: These results suggest that the ablation of TRPC6 may mitigate renal fibrosis by inhibiting the apoptosis and inflammation of PTECs through down-regulation of the PI3K/AKT/GSK3ß pathway. Targeting TRPC6 could be a novel therapeutic strategy for preventing chronic kidney disease.
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Fibrosis , Glucógeno Sintasa Quinasa 3 beta , Ratones Noqueados , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Transducción de Señal , Canal Catiónico TRPC6 , Animales , Canal Catiónico TRPC6/genética , Canal Catiónico TRPC6/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Glucógeno Sintasa Quinasa 3 beta/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Ratones , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Factor de Crecimiento Transformador beta1/metabolismo , Factor de Crecimiento Transformador beta1/genética , Masculino , Riñón/patología , Riñón/metabolismo , Enfermedades Renales/metabolismo , Enfermedades Renales/genética , Enfermedades Renales/patología , Enfermedades Renales/etiología , Células Epiteliales/metabolismo , Células Epiteliales/patología , ApoptosisRESUMEN
BACKGROUND AND OBJECTIVE: Interleukin-6 (IL-6) is the critical factor of early warning, monitoring, and prognosis in the inflammatory storm of COVID-19 cases. IL-6 inducing peptides, which can induce cytokine IL-6 production, are very important for the development of diagnosis and immunotherapy. Although the existing methods have some success in predicting IL-6 inducing peptides, there is still room for improvement in the performance of these models in practical application. METHODS: In this study, we proposed UsIL-6, a high-performance bioinformatics tool for identifying IL-6 inducing peptides. First, we extracted five groups of physicochemical properties and sequence structural information from IL-6 inducing peptide sequences, and obtained a 636-dimensional feature vector, we also employed NearMiss3 undersampling method and normalization method StandardScaler to process the data. Then, a 40-dimensional optimal feature vector was obtained by Boruta feature selection method. Finally, we combined this feature vector with extreme randomization tree classifier to build the final model UsIL-6. RESULTS: The AUC value of UsIL-6 on the independent test dataset was 0.87, and the BACC value was 0.808, which indicated that UsIL-6 had better performance than the existing methods in IL-6 inducing peptide recognition. CONCLUSIONS: The performance comparison on independent test dataset confirmed that UsIL-6 could achieve the highest performance, best robustness, and most excellent generalization ability. We hope that UsIL-6 will become a valuable method to identify, annotate and characterize new IL-6 inducing peptides.
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Biología Computacional , Interleucina-6 , Péptidos , Humanos , Péptidos/química , Biología Computacional/métodos , COVID-19 , Algoritmos , Aprendizaje Automático , SARS-CoV-2RESUMEN
Corneal alkali burns often occur in industrial production and daily life, combined with infection, and may cause severe eye disease. Oxidative stress and neovascularization (NV) are important factors leading to a poor prognosis. URP20 is an antimicrobial peptide that has been proven to treat bacterial keratitis in rats through antibacterial and anti-NV effects. Therefore, in this study, the protective effect and influence mechanism of URP20 were explored in a rat model of alkali burn together with pathogenic bacteria (Staphylococcus aureus and Escherichia coli) infection. In addition, human umbilical vein endothelial cells (HUVECs) and human corneal epithelial cells (HCECs) were selected to verify the effects of URP20 on vascularization and oxidative stress. The results showed that URP20 treatment could protect corneal tissue, reduce corneal turbidity, and reduce the NV pathological score. Furthermore, URP20 significantly inhibited the expression of the vascularization marker proteins VEGFR2 and CD31. URP20 also reduced the migration ability of HUVECs. In terms of oxidative stress, URP20 significantly upregulated SOD and GSH contents in corneal tissue and HCECs (treated with 200 µM H2O2) and promoted the expression of the antioxidant protein Nrf2/HO-1. At the same time, MDA and ROS levels were also inhibited. In conclusion, URP20 could improve corneal injury combined with bacterial infection in rats caused by alkali burns through antibacterial, anti-NV, and antioxidant activities.
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Infecciones Bacterianas , Quemaduras Químicas , Lesiones de la Cornea , Neovascularización de la Córnea , Quemaduras Oculares , Ratas , Humanos , Animales , Quemaduras Químicas/complicaciones , Quemaduras Químicas/tratamiento farmacológico , Quemaduras Químicas/metabolismo , Neovascularización de la Córnea/metabolismo , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Peróxido de Hidrógeno/farmacología , Neovascularización Patológica/metabolismo , Lesiones de la Cornea/tratamiento farmacológico , Células Endoteliales de la Vena Umbilical Humana , Antibacterianos/uso terapéutico , Antibacterianos/farmacología , Quemaduras Oculares/inducido químicamente , Quemaduras Oculares/tratamiento farmacológico , Quemaduras Oculares/patología , Modelos Animales de Enfermedad , Álcalis/toxicidadRESUMEN
Diabetic nephropathy (DN) is a renal microvascular complication caused by diabetes mellitus. One of the most typical characteristics of DN is glomerular mesangial cells (GMCs) proliferation. Stromal interaction molecule 1 (STIM1), a Ca2+ channel, is involved in many diseases. In this study, we investigated the role of STIM1 in the proliferation and fibrosis in high glucose (HG)-induced HBZY-1 cells. We found that the expression of STIM1 was increased in renal tissues of diabetic rat and HBZY-1 cells stimulated by HG. Downregulation of STIM1-mediated SOCE suppressed hyperglycemic cell proliferation and fibrosis by activating autophagy. In addition, the inhibitory effect of downregulating STIM1 on cells was blocked by autophagy inhibitor Bafilomycin A1 (BafA1). Moreover, this experiment also showed that STIM1 regulated autophagy, cell proliferation and fibrosis via PI3K/AKT/mTOR signal pathway. These results clarify the role of STIM1 in HBZY-1 cells and its mechanism, and provide a new target for the treatment of DN.
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AIM: To introduce and evaluate the clinical efficacy of a new technique, the use of viscoelastic substances (VS) to close leaking sclerotomy in 23G microincision vitrectomy, and to observe its effect on the visual acuity and intraocular pressure (IOP) of patients. METHODS: Patients who underwent 23G vitrectomy in Ningbo Eye Hospital before the use of VS technique (June 2019 to September 2020) and after the use of VS technique (October 2020 to December 2021) were selected as the subjects of this study. The above cases underwent operation by the same surgeon and were retrospectively analyzed. VS technique was used as the alternative to suturing, in which a small amount of VS was injected at the leaking sclerotomy and then gently massaged to confirm leaking sclerotomy closure. RESULTS: A total of 174 eyes were covered in the study, including 84 eyes in the control group (before the use of VS technique) and 90 eyes in the VS technique group. The number of eyes that needed to be sutured decreased considerably from 42.9% in the control group to 3.3% in the VS technique group, and the proportion of subconjunctival hemorrhage at 1-2d after surgery decreased remarkably from 35.7% in the control group to 2.2% in the VS technique group. No substantial differences in the incidence of mean IOP and low IOP were found between 1-2 and 3-20d after surgery in the VS technique group. No major complications associated with VS technique were identified during the study. CONCLUSION: In 23G microincision vitrectomy, VS technique is a safe, simple, and effective method to close leaking sclerotomy.
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AIM: To investigate whether nintedanib can inhibit pterygium cells through the fibroblast growth factor receptor 2 (FGFR2)/extracellular-signal-regulated kinase (ERK) pathway. METHODS: Human primary pterygium cells were cultured in vitro. After treatment with nintedanib, the cell morphology was observed under microscopy, the morphological changes of the nucleus were observed after DAPI staining, apoptosis was analyzed by Annexin-V FITC/PI double staining, and the changes of apoptosis-associated proteins were detected by Western blot. The binding ability of nintedanib to FGFR2 was predicted by molecular docking. Finally, by silencing FGFR2, we explored whether nintedanib inhibited FGFR2/ERK pathway. RESULTS: The results showed that nintedanib inhibited the growth of pterygium cells and caused nuclear pyknosis. The results of Annexin-VFITC/PI double staining showed that nintedanib was able to induce early and late apoptosis of pterygium cells, significantly increasing the expression of apoptosis-associated proteins Bax and cleaved-Caspase3 (P<0.05), and reducing the expression of Bcl-2 (P<0.05). In addition, nintedanib significantly inhibited ERK1/2 phosphorylation through FGFR2 (P<0.05). After silencing the expression of FGFR2, there was no significant difference in the inhibition of ERK1/2 phosphorylation by nintedanib (P>0.05). CONCLUSION: Nintedanib induces apoptosis of pterygium cells by inhibiting FGFR2/ERK pathway.
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Due to the global outbreak of COVID-19 and its variants, antiviral peptides with anti-coronavirus activity (ACVPs) represent a promising new drug candidate for the treatment of coronavirus infection. At present, several computational tools have been developed to identify ACVPs, but the overall prediction performance is still not enough to meet the actual therapeutic application. In this study, we constructed an efficient and reliable prediction model PACVP (Prediction of Anti-CoronaVirus Peptides) for identifying ACVPs based on effective feature representation and a two-layer stacking learning framework. In the first layer, we use nine feature encoding methods with different feature representation angles to characterize the rich sequence information and fuse them into a feature matrix. Secondly, data normalization and unbalanced data processing are carried out. Next, 12 baseline models are constructed by combining three feature selection methods and four machine learning classification algorithms. In the second layer, we input the optimal probability features into the logistic regression algorithm (LR) to train the final model PACVP. The experiments show that PACVP achieves favorable prediction performance on independent test dataset, with ACC of 0.9208 and AUC of 0.9465. We hope that PACVP will become a useful method for identifying, annotating and characterizing novel ACVPs.
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COVID-19 , Péptidos , Humanos , Algoritmos , Aprendizaje Automático , ProbabilidadRESUMEN
According to the latest data from the Bureau of Disease Control and Prevention of the National Health and Family Planning Commission, China currently has 199.6 million diabetic patients and has become the world's largest country with diabetes. The prevalence rate is as high as 14.3%, which is much higher than the world average of 5.8%. The primary-level ophthalmic screening service is one of the important tasks to improve primary-level medical services, and the corresponding ophthalmic imaging diagnosis technology is an important support for primary-level medical and health services. Therefore, it is very necessary for us to study the application of artificial intelligence image recognition technology for diabetic retinopathy under the medical consortium mode and to study the precise initial diagnosis, precise referral, and precise follow-up of diabetic retina under the medical conjoined mode, so as to better promote the transformation of the ophthalmology primary service model. Based on this background, in this article, we have proposed and carried out the following solution: (1) diabetes data collation. Based on medical artificial intelligence technology, this paper collected 2,265 electronic medical records from an eye hospital in Ningbo and selected 2,000 qualified medical records for data integration and preprocessing. The contents of electronic medical records mainly include age, gender, and examination records. (2) Establish diabetic retinopathy diagnosis model based on neural network algorithm. This article first uses the classic algorithm of BP neural network for modeling, chooses the Levenberg-Marquardt method as the training function, and selects 10 hidden layer units through comparison experiments. After that, ophthalmologists assessed 80 sets of test results and determined the right diagnosis rate. Finally, this article compares and analyzes the accuracy of the two routes in 80 tests.
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Diabetes Mellitus , Retinopatía Diabética , Inteligencia Artificial , Retinopatía Diabética/diagnóstico por imagen , Humanos , Tamizaje Masivo , Redes Neurales de la Computación , RetinaRESUMEN
Intravitreal anti-VEGF (anti-vascular endothelial growth factor) biologics have revolutionized the pharmacological management of chorioretinal diseases. However, the systemic adverse events such as stroke or bleeding are the concerns for many patients and physicians. The mechanism to develop these side effects are poorly understood. Consecutive 95 patients with retinal diseases were studied for their blood activated partial thromboplastin time (APTT), prothrombin time (PT), international normalized ratio (INR), and concentration of fibrinogen before and after intravitreal conbercept. Additionally, plasma nitric oxide (NO) and endothelin-1 (ET-1) were investigated on 38 of the 95 patients. Compared with the pre-injection, 4-week post-injection values of APTT and PT were increased by 0.582 s (p = 0.038, paired t test) and by 0.086 s (p = 0.080, paired t test; p = 0.0475, Sign test), respectively. At the same time, fibrinogen decreased by 0.048 g/L. Plasma levels of NO or ET-1 or VEGF did not significantly change from pre-injection levels. Our findings advanced the understanding of mechanism for systemic side effects associated with intravitreal anti-VEGF and emphasized paying more attention to higher risk of possible bleedings for patients following intravitreal conbercept.
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Coagulación Sanguínea/efectos de los fármacos , Endotelina-1/sangre , Óxido Nítrico/sangre , Proteínas Recombinantes de Fusión/efectos adversos , Enfermedades de la Retina/tratamiento farmacológico , Anciano , Femenino , Fibrinógeno/metabolismo , Humanos , Inyecciones Intravítreas , Masculino , Persona de Mediana Edad , Protrombina/metabolismo , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/uso terapéutico , Tromboplastina/metabolismoRESUMEN
Renal ischemia/reperfusion (I/R), a major cause of acute kidney injury (AKI), is a serious clinical event in patients during post-renal transplantation. I/R is associated with renal dysfunction and tubular apoptosis, and calcium (Ca2+) overload has been reported to be a crucial factor on tubular apoptosis in I/R injury (IRI). The canonical transient receptor potential channel 6 (TRPC6), a type of non-selective Ca2+ channel, is involved in many renal diseases. Our earlier study identified that TRPC6-mediated Ca2+ influx plays a novel role in suppressing cytoprotective autophagy triggered by oxidative stress in primary tubular epithelial cells (TECs). This study explored the potential beneficial impact of TRPC6 knockout (TRPC6-/-) and the relevant cellular mechanisms against I/R-induced AKI in mice. Measuring changes of renal function, apoptotic index, and autophagy in mouse kidneys that suffered 24 h reperfusion after 40 min ischemia and working in vitro with TECs that suffered 24 h reoxygenation after 24 h hypoxia, we found that 1) IRI tissues had increased TRPC6 expression and TRPC6 knockout significantly ameliorated renal damage induced by IRI; 2) TRPC6 knockout enhanced the level of autophagy and alleviated the depolarization of mitochondrial membrane potential (ψm, MMP) and apoptotic changes upon IRI; and 3) IRI tissues had increased p-AKT and p-ERK1/2 expressions, while TRPC6 knockout could markedly reduce the phosphorylation of AKT and ERK1/2. These discoveries suggest that, by reducing Ca2+ overload, the underlying protective mechanism of TRPC6-/- may be involved in down-regulation of PI3K/AKT and ERK signaling, which is likely to provide a new avenue for future AKI therapies.
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BACKGROUND: Age-related macular degeneration (AMD) is one of the major causes of blindness, and the incidence of this disease has been increasing in recent years. OBJECTIVE: To investigate the association between the single nucleotide polymorphisms (SNPs) of the high temperature requirement factor A-1 (HTRA1) and complement factor H (CFH) genes and susceptibility to AMD in Ningbo, China. METHODS: Ninety-eight patients with AMD and seventy-three controls were recruited at the Sixth Hospital of Ningbo from August 2017 to April 2019 in China. Genomic DNA was extracted from the venous blood provided by the hospital, and the genotypes of the AMD susceptibility genes CFH and HTAR1 were detected by polymerase chain reaction and sequenced directly. The SNPs rs11200638 on the HTRA1 gene and rs3753394 on the CFH gene were selected for genotype and association analysis. The correlations between the different genotypes of HTRA1 and CFH and AMD were analysed by the Chi-squared test. RESULTS: All the genotypes adhered to the Hardy-Weinberg equilibrium. There were three genotypes (AA, AG and GG) in HTRA1 (rs11200638). The differences in genotypes and allele frequency between the AMD group and the control group were statistically significant (P < 0.05). The A allele was a risk allele (OR: 4.19, 95% Cl: 2.28 ~ 7.70, P < 0.05), with a frequency of 61.7% in patients versus 43.8% in controls. However, the rs3753394 SNP in CFH was not associated with AMD in our study (P > 0.05). CONCLUSIONS: The rs11200638 SNP of the HTRA1 gene is associated with AMD, and the AA genotype is a risk factor for AMD in the Ningbo population. There is no significant correlation between the rs3753394 SNP of the CFH gene and AMD.
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Factor H de Complemento , Degeneración Macular , China/epidemiología , Factor H de Complemento/genética , Frecuencia de los Genes , Genotipo , Serina Peptidasa A1 que Requiere Temperaturas Altas/genética , Humanos , Lactante , Degeneración Macular/genética , Polimorfismo de Nucleótido Simple , Serina Endopeptidasas/genética , Vitamina B 12/análogos & derivadosRESUMEN
OBJECTIVE: This study explores the correlations of stromal cell-derived factor-1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) levels with caspase-3 expression in the retina of rats after optic nerve injury. MATERIALS AND METHODS: A total of 24 adult healthy specific pathogen-free Sprague-Dawley rats were selected and randomly divided into an optic nerve injury group (n=16) and an optic nerve sham-injury group (n=8). The optic nerve injury group was further sub-divided into a 3 d group (n=8) and a 7 d group (n=8) after their injuries. In the optic nerve injury group, the left eye of each rat was removed and prepared for the optic nerve injury model using the optic nerve clamping method. In the sham-injury group, the optic nerve in the left eye was only exposed without being clamped. The rats were sacrificed at 3 d and 7 d after their optic nerve injuries, and the retina was isolated. The expressions of SDF-1, CXCR4, and caspase-3 in the retina of the rats in each group were measured using an immunohistochemical method. Messenger ribonucleic acid (mRNA) and the protein expressions of SDF-1, CXCR4, and caspase-3 (cleaved caspase-3) in the retinas of rats were measured using quantitative real-time polymerase chain reaction (qPCR) and western blotting, respectively. Moreover, the correlations of the expression of SDF-1 and CXCR4 with caspase-3 expression were analyzed using the Spearman method. The apoptosis of retinal ganglion cells of rats in each group was observed using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. RESULTS: Immunohistochemistry of the retinas revealed that, compared with those in the sham-injury group, the expressions of SDF-1 and CXCR4 in the retina in the 3 d group and the 7 d group were gradually increased. Caspase-3 expression was significantly elevated at 3 d after the injuries, but obviously decreased at 7 d after the injuries. The results of qPCR showed that the relative expression levels of SDF-1 and CXCR4 mRNA in the retina at 3 d and 7 d after optic nerve injuries were also significantly higher than those in the sham-injury group (P<0.01), and the caspase-3 mRNA expression was initially increased at 3 d but reduced at 7 d after the injuries (P<0.01). Western blotting for the detection of the SDF-1, CXCR4 and caspase-3 proteins indicated changes similar to those of the qPCR. Spearman analysis results demonstrated that there was a positive correlation between the SDF-1 and CXCR4 expressions, but the expressions of SDF-1 and CXCR4 had negative correlations with caspase-3 expression. TUNEL staining showed that apoptosis of the retinal cells was increased in the 3 d group but significantly decreased in the 7 d group. CONCLUSION: After optic nerve injury, the continuous increase of the SDF-1 and CXCR4 levels suppresses the apoptosis of retinal cells and repairs the retina by inhibiting the cleavage activation of caspase-3. This provides new insights for the ongoing treatment of optic nerve injury.
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High-conductance, voltage- and calcium-activated potassium channels (BKCa) and store-operated calcium channels (SOCs) are belong to K+ channels superfamily and calcium channels superfamily respectively. Since BKCa potassium channels can be activated by calcium ions, we set out to examine whether SOCs are coupled with BKCa and to probe the relationship of BKCa and SOCs. First, we proved that BKCa immunoprecipitated with Orai1, and confocal microscopy showed that BKCa co-localized with Orai1. Next, we mapped that the exact binding sites locate in the 350aa-371aa fragment of the first regulatory domain associated with K+ conduction (RCK1) and the 720aa-814aa fragment in the second regulatory domain associated with K+ conduction (RCK2) via GST-pull-down assays. Then, we showed, by calcium imaging that BKCa overexpression enhanced endogenous and exogenous store-operated calcium entry (SOCE) and this enhancement could be blocked by Orai1 knockdown. Finally, we proved that SOCE could enhance the activity of BKCa by patch-clamp. From these results we conclude that BKCa can form a positive feedback loop with SOCs, as the Ca2+ influx from SOCs can active BKCa, which can hyperpolarize the plasma membrane. In turn, the hyperpolarized membrane will form a higher electric potential difference and give more force, allowing Ca2+ influx via SOCs.
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Subunidades alfa de los Canales de Potasio de Gran Conductancia Activados por Calcio/metabolismo , Proteína ORAI1/metabolismo , Sitios de Unión , Calcio/metabolismo , Células HEK293 , Humanos , Unión Proteica/efectos de los fármacos , Mapeo de Interacción de Proteínas , Molécula de Interacción Estromal 1/metabolismo , Tapsigargina/farmacologíaRESUMEN
AIM: To investigate the effects of nintedanib thermo-sensitive hydrogel (NTH) on neovascularization and related markers in corneal alkali burns of Wistar rats. METHODS: NTH was prepared by grinding, and its phase-transition temperature was determined. Thirty specific-pathogen-free Wistar rats served as a model of corneal alkali burn in the right eye were randomly divided into 3 groups (n=10, each): model group treated with 0.9% saline once a day, NTH group with 0.2% nintedanib b.i.d, and dexamethasone group with dexamethasone ointment once a day. The left eye of rats served as the controls. The corneal transparency was observed under a slit-lamp microscope, and the area of neovascularization was calculated. On day 7, the rats were sacrificed, and the cornea was removed and embedded with paraffin, then stained with hematoxylin-eosin, and the expression of vascular endothelial growth factor receptor 2 (VEGFR-2) and CD31 in the corneal tissues of each group was detected by immunofluorescence. RESULTS: The phase-transition temperature of nintedanib obtained by grinding was 37°C after adding artificial tears. The results of the alkali burn model indicated that the growth rate of neovascularization in the NTH group was slower than that in the model group, and the neovascularization area was significantly smaller than that in the model group (P<0.05). Moreover, CD31 and VEGFR-2 expression levels in the NTH group were significantly lower than those in the model group. CONCLUSION: NTH becomes colloidal at body temperature, which is beneficial for releasing the drug slowly and can significantly inhibit the neovascularization of corneal induced by alkali burn in rats.
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Kidney fibrosis is generally confirmed to have a significant role in chronic kidney disease, resulting in end-stage kidney failure. Epithelial-mesenchymal transition (EMT) is an important molecular mechanism contributing to fibrosis. Tubular epithelial cells (TEC), the major component of kidney parenchyma, are vulnerable to different types of injuries and are a significant source of myofibroblast by EMT. Furthermore, TRPC6 knockout plays an anti-fibrotic role in ameliorating kidney damage. However, the relationship between TRPC6 and EMT is unknown. In this study, TRPC6-/- and wild-type (WT) mice were subjected to a unilateral ureteric obstruction (UUO) operation. Primary TEC were treated with TGF-ß1. Western blot and immunofluorescence data showed that fibrotic injuries alleviated with the inhibition of EMT in TRPC6-/- mice compared to WT mice. The activation of AKT-mTOR and ERK1/2 pathways was down-regulated in the TRPC6-/- mice, while the loss of Na+/K+-ATPase and APQ1 was partially recovered. We conclude that TRPC6 knockout may ameliorate kidney fibrosis by inhibition of EMT through down-regulating the AKT-mTOR and ERK1/2 pathways. This could contribute to the development of effective therapeutic strategies on chronic kidney diseases.
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BACKGROUND: The effectiveness of folic acid supplementation in stroke risk has been investigated, however, the available results are inconclusive and conflicting. The purpose of this systemic review and meta-analysis was to assess the effect of folic acid in patients with cardiovascular disease (CVD). METHODS: By searching the PubMed, EMBASE, and Cochrane library databases, we conducted a meta-analysis to evaluate effect of folic acid supplementation in patients with CVD. All-cause mortality, cardiovascular mortality, the risk of coronary heart disease (CHD) and stroke were summarized; hazard ratios (HR), the relative risk (RR) and its 95% confidence interval (CI) were also calculated. Fixed effects models were used to combine the data. A total of 12 randomized controlled trials, which involved 47,523 participants, met the inclusion criteria in this systematic review and meta-analysis. RESULTS: Our meta-analysis showed that cardiovascular patients who received folic acid therapy had significantly decreased risk of stroke (RRâ=â0.85, 95% CIâ=â0.77-0.94, Pheterogeneityâ=â.347, Iâ=â10.6%) compared with patients who received control treatment. However, no significant difference in all-cause mortality (HR, 0.97, 95% CI, 0.86-1.10, Pheterogeneityâ=â.315, Iâ=â15.4%), cardiovascular mortality (HR, 0.87, 95% CI, 0.66-1.15, Pheterogeneityâ=â.567, Iâ=â0) and risk of CHD (RR, 1.04, 95% CI, 0.99-1.10, Pheterogeneityâ=â.725, Iâ=â0) were found between the 2 groups. CONCLUSION: This meta-analysis suggested that folic acid supplementation significantly reduced the risk of stroke in patients with CVD.
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Enfermedades Cardiovasculares/tratamiento farmacológico , Ácido Fólico/farmacología , Enfermedades Cardiovasculares/mortalidad , Suplementos Dietéticos , Ácido Fólico/uso terapéutico , Humanos , Complejo Vitamínico B/farmacología , Complejo Vitamínico B/uso terapéuticoRESUMEN
Tris (1, 3-dichloro-2-propyl) phosphate (TDCPP) is a major type of organophosphorus flame retardants, and long-term exposure to TDCPP to normal cells or tissues under physiological conditions can induce toxic effects. But how TDCPP leads to the adverse effects is not yet clear, and the effect of TDCPP under pathological conditions such as reactive oxygen species assault is not well understood. The present study aimed to explore the potential effect of TDCPP against H2O2-induced oxidative stress in H9c2 cardiomyoblasts and rat neonatal cardiomyocytes. We found that H2O2-treatment decreased cell viability and increased lactate dehydrogenase and malondialdehyde generation of H9c2 cells. However, TDCPP could alleviate these effects. TDCPP alleviated Ca2+-overload caused by H2O2 through decreasing store-operated calcium entry. More importantly, TDCPP remarkably decreased H2O2-induced dephosphorylation of Akt and GSK3ß, and through this pathway TDCPP mitigated the H2O2-induced apoptosis and detrimental autophagy. Collectively, via mitigating Ca2+-overload and activating the Akt/GSK3ß signaling pathway, TDCPP may have a role in protecting cardiomyocytes from oxidative stress.
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Señalización del Calcio/efectos de los fármacos , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Peróxido de Hidrógeno/farmacología , Miocitos Cardíacos/metabolismo , Compuestos Organofosforados/farmacología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Calcio/metabolismo , Línea Celular , Humanos , Ratones , Miocitos Cardíacos/patología , Estrés Oxidativo/efectos de los fármacos , Fosforilación/efectos de los fármacos , RatasRESUMEN
Reactive oxygen species (ROS) are generated under various pathological conditions such as renal ischemia/reperfusion (I/R) injury and provoke damage to multiple cellular organelles and processes. Overproduction of ROS causes oxidative stress and contributes to damages of renal proximal tubular cells (PTC), which are the main cause of the pathogenesis of renal I/R injury. Autophagy is a dynamic process that removes long-lived proteins and damaged organelles via lysosome-mediated degradation, which has an antioxidant effect that relieves oxidative stress. The canonical transient receptor potential channel 6 (TRPC6), a nonselective cation channel that allows passage of Ca2+, plays an important role in renal disease. Yet, the relationship between TRPC6 and autophagy, as well as their functions in renal oxidative stress injury, remains unclear. In this study, we found that oxidative stress triggered TRPC6-dependent Ca2+ influx in PTC to inhibit autophagy, thereby rendering cells more susceptible to death. We also demonstrated that TRPC6 knockout (TRPC6-/-) or inhibition by SAR7334, a TRPC6-selective inhibitor, increased autophagic flux and mitigated oxidative stress-induced apoptosis of PTC. The protective effects of TRPC6 ablation were prevented by autophagy inhibitors Chloroquine and Bafilomycin A1. Moreover, this study also shows that TRPC6 blockage promotes autophagic flux via inhibiting the PI3K/Akt/mTOR and ERK1/2 signaling pathways. This is the first evidence showing that TRPC6-mediated Ca2+ influx plays a novel role in suppressing cytoprotective autophagy triggered by oxidative stress in PTC, and it may become a novel therapeutic target for the treatment of renal oxidative stress injury in the future.
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Apoptosis/genética , Autofagia/genética , Células Epiteliales/patología , Túbulos Renales Proximales/patología , Estrés Oxidativo/genética , Canal Catiónico TRPC6/genética , Animales , Antioxidantes/metabolismo , Calcio/metabolismo , Señalización del Calcio/genética , Línea Celular Tumoral , Células Epiteliales/metabolismo , Humanos , Enfermedades Renales/genética , Enfermedades Renales/metabolismo , Enfermedades Renales/patología , Túbulos Renales Proximales/metabolismo , Lisosomas/genética , Lisosomas/metabolismo , Lisosomas/patología , Sistema de Señalización de MAP Quinasas/genética , Masculino , Ratones , Ratones Noqueados , Fosfatidilinositol 3-Quinasas/genética , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/genética , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Transducción de Señal/genética , Serina-Treonina Quinasas TOR/genéticaRESUMEN
TDCPP, Tris (1, 3-dichloro-2-propyl) phosphate belongs to a group of chemicals known as triester organophosphate flame retardants, It can alter calcium homeostasis at much lower concentrations in normal conditions, but the mechanism is unclear till now. Calcium overload is a leading cause of apoptosis in myocardial ischemia/reperfusion (I/R) injury, thus how to mitigate Ca2+-overload is deserved to be investigated. We therefore hypothesized that TDCPP could attenuate cardiomyocytes apoptosis in I/R injury. H/R (hypoxia/reoxygenation) experiments in vitro were used to simulate in vivo I/R injury. The present study aimed to explore the potential effect of TDCPP in cardiomyocytes after H/R injury, Ca2+ imaging technique was used to explore SOCE(store-operated calcium entry) and Ca2+-overload levels, western blot technique was used to explore the potential target, the cell morphology, cell viability and mitochondrial membrane potential were also detected. The results have shown that: TDCPP could decrease SOCE, restore H9c2 cell viability, mitigate Ca2+-overload in H/R injury and reduce the mitochondrial membrane potential. Furthermore, TDCPP decreased STIM1 expression and promoted GSK3ß phosphorylation. Collectively, for the first time, this study suggest the antiapoptosis roles of TDCPP in H/R injury are via mitigation Ca2+-overload and promoting GSK-3ß phosphorylation.
Asunto(s)
Apoptosis/efectos de los fármacos , Calcio/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Hipoxia/tratamiento farmacológico , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Miocitos Cardíacos/efectos de los fármacos , Compuestos Organofosforados/farmacología , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Hipoxia/metabolismo , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Daño por Reperfusión Miocárdica/metabolismo , Miocitos Cardíacos/metabolismo , Fosforilación/efectos de los fármacos , Sustancias Protectoras/farmacología , Ratas , Molécula de Interacción Estromal 1/metabolismoRESUMEN
BACKGROUND: Cervical cancer is a major cause of death in women worldwide. Interferon-induced transmembrane protein 1 (IFITM1) is involved in antivirus defense, cell adhesion, and carcinogenesis in different tissues. However, the role of IFITM1 gene in cervical squamous cell cancer is unclear. METHODS: To explore the role of IFITM1 in carcinogenesis of cervical cancer, we investigated the expression of IFITM1 gene in cervical squamous cell carcinoma. IFITM1 mRNA level was measured by real-time quantitative RT-PCR in cervical cancer tissues and their adjacent normal tissues. IFITM1 protein level was measured by immunohistochemistry. Methylation in the IFITM1 gene promoter was detected by methylation-specific PCR. We then transfected HeLa cells with IFITM1 expression vector or control vector. IFITM1 expression was examined; cell migration and invasion were analyzed by wound healing assay and matrigel-coated transwell migration assays, respectively. HeLa cell proliferation was measured by cell counting kit-8 assay and cell cycle analysis. Cell apoptosis was analyzed by Annexin V/propidium iodide double staining assay. RESULTS: The difference in IFITM1 protein expression between samples from chronic cervicitis and cervical carcinoma was statistically significant (P < 0.01). Ki-67 and PCNA protein expression levels were significantly higher in cervical cancer tissues than in their corresponding cervicitis tissues (P < 0.05 and P < 0.001, respectively). IFITM1 mRNA level was significantly lower in cervical cancer tissues than in normal cervical tissues (P < 0.05). Methylation of the IFITM1 gene promoter was significantly higher in cervical cancer than in normal cervical tissues (P < 0.05). Transfection of the IFITM1 pcDNA3.1 construct decreased cell migration and invasion of HeLa cells, inhibited cell proliferation, and increased cell apoptosis. CONCLUSION: IFITM1 gene expression may reduce the proliferation, migration, and invasion of cervical squamous cancer cells.