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Four trials were conducted to establish a protein and amino acid requirement model for layer chicks over 0-6 weeks by using the analytical factorization method. In trial 1, a total of 90 one-day-old Jing Tint 6 chicks with similar body weight were selected to determine the growth curve, carcass and feather protein deposition, and amino acid patterns of carcass and feather proteins. In trials 2 and 3, 24 seven-day-old and 24 thirty-five-day-old Jing Tint 6 chicks were selected to determine the protein maintenance requirements, amino acid pattern, and net protein utilization rate. In trial 4, 24 ten-day-old and 24 thirty-eight-day-old Jing Tint 6 chicks were selected to determine the standard terminal ileal digestibility of amino acids. The chicks were fed either a corn-soybean basal diet, a low nitrogen diet, or a nitrogen-free diet throughout the different trials. The Gompertz equation showed that there is a functional relationship between body weight and age, described as BWt(g) = 2669.317 × exp(-4.337 × exp(-0.019t)). Integration of the test results gave a comprehensive dynamic model equation that could accurately calculate the weekly protein and amino acid requirements of the layer chicks. By applying the model, it was found that the protein requirements for Jing Tint 6 chicks during the 6-week period were 21.15, 20.54, 18.26, 18.77, 17.79, and 16.51, respectively. The model-predicted amino acid requirements for Jing Tint 6 chicks during the 6-week period were as follows: Aspartic acid (0.992-1.284), Threonine (0.601-0.750), Serine (0.984-1.542), Glutamic acid (1.661-1.925), Glycine (0.992-1.227), Alanine (0.909-0.961), Valine (0.773-1.121), Cystine (0.843-1.347), Methionine (0.210-0.267), Isoleucine (0.590-0.715), Leucine (0.977-1.208), Tyrosine (0.362-0.504), Phenylalanine (0.584-0.786), Histidine (0.169-0.250), Lysine (0.3999-0.500), Arginine (0.824-1.147), Proline (1.114-1.684), and Tryptophan (0.063-0.098). In conclusion, this study constructed a dynamic model for the protein and amino acid requirements of Jing Tint 6 chicks during the brooding period, providing an important insight to improve precise feeding for layer chicks through this dynamic model calculation.
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[This corrects the article DOI: 10.3892/ol.2019.10694.].
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This meta-analysis aims to systemically evaluate the efficacy of vacuum sealing drainage (VSD) combined with autologous platelet-rich plasma (PRP) in the treatment of diabetic foot ulcers (DFU). The China HowNet, China Biomedical Literature, VIP periodical resource integration service platform, Wanfang, Embase, Cochrane Central, and PubMed databases were retrieved using the computer. The retrieval period was up to July 2021. Randomised controlled trials on VSD combined with PRP in the treatment of DFU were collected. Those trials that met the inclusion criteria were included for meta-analysis using RevMan 5.3 software. A total of 13 articles were included. In the trial group, 477 patients with DFU were treated with VSD combined with PRP, while in the control group, 482 patients with DFU were treated with conventional dressings and/or VSD. The meta-analysis showed that, compared with the control group, VSD combined with PRP has significant advantages in shortening healing time (standardised mean difference [SMD] = -0.87, 95% confidence interval [CI]: -1.07 to -0.67, P < .00001), improving ulcer healing rates (odds ratio = 4.01, 95% CI: 2.95 ~ 5.46, P < .00001), and reducing hospital stays (mean difference = -15.29, 95% CI: -16.05 to -14.54, P < .00001), but the differences in dressing change times (SMD = -1.27, 95% CI: -2.71 to 0.17, P = .08) and hospitalisation expenses (SMD = -0.16, 95% CI: -13.40 to 13.07, P = .98) were not statistically significant. VSD combined with autologous PRP has good curative efficacy in the treatment of DFU and is a better treatment option. However, this treatment is limited in patients with platelet dysfunction, thrombocytopenia, leukaemia, and poor general condition.
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Diabetes Mellitus , Pé Diabético , Tratamento de Ferimentos com Pressão Negativa , Plasma Rico em Plaquetas , Humanos , Pé Diabético/terapia , Drenagem , CicatrizaçãoRESUMO
Objective: The aim was to systematically compare the drug compatibility with various closed intravenous (i.v.) infusion containers, to provide a reference for selecting a relatively superior infusion container and improve the medication safety for patients in clinical practice. Methods: The compatibility of four commonly used clinical injections (ceftazidime, pantoprazole sodium, ambroxol hydrochloride, edaravone) with three representative closed i. v. infusion containers (non-PVC infusion bags, upright polypropylene infusion bags, inner sealed polypropylene infusion bags) prefilled with infusion fluids (0.9% sodium chloride or 5% dextrose) in the Chinese market were investigated in this study. The particle counts of both infusion fluids and diluted chemical injections by infusion fluids in various infusion containers were determined by the light obscuration method. At 0, 2 and 6 h after four injections following dilution with infusion fluids in each container, the pH of the solutions was detected, and the physical properties were examined by visual inspection. Meanwhile, the drug concentrations were assessed by high performance liquid chromatography (HPLC). Results: As for either infusion fluids or diluted injections by infusion fluids, the particle counts in non-PVC infusion bags were significantly greater than those in the other two bags under some circumstances. The particle counts in diluted injections by infusion fluids increased dramatically compared with those in infusion fluids in all infusion containers, especially for the small-size particles. But pH, physical properties and drug concentrations of diluted infusion solutions in all infusion containers remained nearly unchanged over the test period. Conclusion: Closed i. v. infusion containers included in this study are all well-compatible with four injections. Moreover, the closed infusion containers produced by Chinese manufacturers have met the international quality standard. Particularly, the intravenous admixture preparation process needs to be optimized to reduce the overall particulate contaminants.
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The proliferation of the biomarker Ki67 has been extensively studied in colorectal cancer (CRC). Although numerous Ki67 cut-off values have previously been reported, the optimal cut-off value remains unclear with previous studies providing contrasting results. The present retrospective cohort study aimed to determine the optimal cut-off value for CRC. Ki67 levels and the prognosis of patients with non-metastatic CRC were obtained from the Electronic Health Information System of a tertiary hospital in Kunming City. The Restricted Cubic Spline (RCS) model was used to analyze the non-linear association between Ki67 levels and the risk of patient death and metastasis. Moreover, the RCS model was used to determine the optimal cut-off value of Ki67. Cox proportional hazards models were used to verify the effects of the cut-off value. In total, 210 patients with CRC and a median age of 62.5 years (age range, 23.0-88.0 years) were studied. Results of the present study demonstrated a non-linear association between Ki67 levels and the risk of patient death based on the RCS model, and at Ki67 levels ≥60%, the hazard ratio (HR) of patient death gradually increased. Using multivariate-adjusted Cox proportional hazards models, the results of the present study demonstrated that Ki67 ≥60% indicated a high-risk ratio for both distant metastasis and death [HR, 2.640; 95% confidence interval (CI), 1.066-6.539], compared with Ki67 <60% (HR, 2.558; 95% CI, 1.079-6.064). Therefore, Ki67 ≥60% may be the optimal cut-off value for the prediction of death and metastasis in patients with CRC. Thus, Ki67 may act as a biomarker for predicting the prognosis of patients with CRC, and the optimal cut-off value for the prediction of both death and metastasis of patients with CRC is 60%.
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Peripheral odontogenic keratocysts are rarely observed, and cases of odontogenic keratocysts of buccal soft tissues are even rarer. This study was performed to present two rare cases of odontogenic keratocysts in buccal soft tissues and review related literature.
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Cistos Odontogênicos , Tumores Odontogênicos , Humanos , Cistos Odontogênicos/diagnósticoRESUMO
Bruton's tyrosine kinase (BTK) is a key component of the B cell receptor (BCR) signaling pathway and plays a crucial role in B cell malignancies and autoimmune disorders; thus, it is an attractive target for the treatment of B cell related diseases. Here, we evaluated the BTK inhibitory activity of a series of pyrimido[4,5-d][1,3]oxazin-2-one derivatives. Combining this evaluation with structure-activity relationship (SAR) analysis, we found that compound 2 exhibited potent BTK kinase inhibitory activity, with an IC50 of 7 nM. This derivative markedly inhibited BTK activation in TMD8 B cell lymphoma cells and thus inhibited the in vitro growth of the cells. Further studies revealed that compound 2 dose dependently arrested TMD8 cells at G1 phase, accompanied by decreased levels of Rb, phosphorylated Rb, and cyclin D1. Moreover, following treatment with compound 2, TMD8 cells underwent apoptosis associated with PARP and caspase 3 cleavage. Interestingly, the results of the kinase activity assay on a small panel of 35 kinases showed that the kinase selectivity of compound 2 was superior to that of the first-generation inhibitor ibrutinib, suggesting that compound 2 could be a second-generation inhibitor of BTK. In conclusion, we identified a potent and highly selective BTK inhibitor worthy of further development.
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Tirosina Quinase da Agamaglobulinemia/antagonistas & inibidores , Antineoplásicos/farmacologia , Descoberta de Drogas , Inibidores de Proteínas Quinases/farmacologia , Pirimidinas/farmacologia , Tirosina Quinase da Agamaglobulinemia/metabolismo , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Pirimidinas/síntese química , Pirimidinas/química , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
A novel Vogesella strain, YM-1T, was recovered from human urine in PR China in 2017. Cells of strain YM-1T were Gram-stain-negative, rod-shaped, aerobic, motile, non-spore-forming and poly-ß-hydroxybutyrate-accumulating. The strain contained C16:1ω6c/C 16:1ω7c, C16:0 and C18:0ω7c as major fatty acids; phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified phospholipid as major polar lipids; and ubiquinone-8 as the predominant respiratory quinone. Comparison of 16S rRNA gene sequences indicated that this strain had highest similarities to Vogesella perlucida DS-28T (98.8â%) and Vogesella mureinivorans 389T (98.1â%). The results of phylogenetic analysis based on the 16S rRNA gene sequences revealed that the novel strain was clustered and well separated with V. perlucida DS-28T and V. mureinivorans 389T within the genus Vogesella. The average nucleotide identity (ANI) and amino acid identity (AAI) analyses showed that this strain was not identified as V. perlucida DS-28T or V. mureinivorans 389T, with values well below the threshold limit for species demarcation (ANI <88.1â%, AAI <88.6â%). Based on the above results, strain YM-1T is proposed to be a novel species of the genus Vogesella with the name Vogesella urethralis sp. nov. (YM-1T=NBRC 113779=CGMCC 1.17135).
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Betaproteobacteria/classificação , Filogenia , Urina/microbiologia , Bactérias Aeróbias/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , Betaproteobacteria/isolamento & purificação , China , DNA Bacteriano/genética , Ácidos Graxos/química , Humanos , Hidroxibutiratos/metabolismo , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Poliésteres/metabolismo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/químicaRESUMO
Ovarian cancer (OC) is a type of gynaecological malignancy with high mortality in females. Serous ovarian cancer (SOC) is a distinct subtype of OC with poor early diagnosis. Given the limitations of traditional therapies, such as chemotherapy, targeted treatment is therefore a promising therapy to improve the survival rate of SOC patients. Cyclophilin A (CYPA) is a member of Cyclophilin family and thought to participates in multiple cellular processes such as cell transduction and immune modulation. Recently, various of studies indicated that CYPA has critical impact on cancer progression. CYPA could regulate cell proliferation, invasion, and chemoresistance of multiple types of cancers. However, it is still unclear whether it could affect ovarian cancer. In this study, we demonstrated that CYPA was highly expressed in SOC tissues compared with adjacent tissues. Further, CYPA was significantly associated with clinical stage and lymphnode metastasis of SOC patients. Additionally, data indicated that knockdown of CYPA by its shRNA dramatically reduces migration and invasion capacity of SOC cells in vitro and blocks tumor metastasis in vivo. Our study investigates the involvement of CYPA in the progression and metastasis of SOC, and therefore provides CYPA as a promising therapeutic target for SOC treatment.
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Ciclofilina A/metabolismo , Cistadenocarcinoma Seroso/metabolismo , Neoplasias Ovarianas/metabolismo , Animais , Linhagem Celular Tumoral , Movimento Celular , Ciclofilina A/genética , Cistadenocarcinoma Seroso/patologia , Progressão da Doença , Feminino , Humanos , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos Nus , Pessoa de Meia-Idade , Neoplasias Ovarianas/patologia , RNA Interferente Pequeno/genéticaRESUMO
The present study focused on exploring the inhibitory mechanism of microRNA (miR)-23a in endometrial cancer. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to investigate miR-23a expression in endometrial tissues and endometrial cancer cells. A colony formation assay using crystal violet staining was performed to compare cell proliferation, while wound-healing and Transwell assays were performed to compare cell migration and invasion. Subsequently, bioinformatics and a luciferase reporter gene assay were used to investigate the effect of miR-23a on sine oculis homeobox homolog 1 (SIX1) expression, and the biological function of SIX1 was analyzed. Additionally, a nude mouse tumorigenicity assay was performed to test the inhibitory effect of miR-23a and Taxol® therapy in endometrial cancer. Finally, immunohistochemistry and RT-qPCR were used to explore the association between miR-23a and SIX1 expression in endometrial cancer tissues. miR-23a was underexpressed in endometrial cancer tissues compared with in para-carcinoma tissues, and the overexpression of miR-23a inhibited proliferation and invasion of endometrial cancer cells. Furthermore, SIX1 was demonstrated to be a downstream target of miR-23a, and miR-23a reduced SIX1 expression. Additionally, SIX1 inversely promoted cell proliferation, migration and invasion. In addition, the effects of reduced cell proliferation and increased cell invasion following miR-23a overexpression could be reversed by adding SIX1 to in vitro culture. Furthermore, the inhibitory effect of miR-23a and Taxol therapy, which reduced SIX1 expression in endometrial cancer, was demonstrated in vivo. Finally, a negative association between miR-23a and SIX1 expression was demonstrated in endometrial cancer tissues. The results of the present study revealed that miR-23a may inhibit endometrial cancer development by targeting SIX1.
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BACKGROUND: Cervical cancer is the most common gynecological malignancy with low terminal cure rate, and therefore new therapeutic targets are urgently needed to combat this disease. SMYD2, as an oncogene, is abnormal highly expressed in multiple types of tumors and further affects the occurrence and development, but the potential correlations between SMYD2 expression and cervical cancer progression is still unclear. METHODS: We first used the bioinformatics website to screen the data of cervical cancer in (The Cancer Genome Atlas) TCGA and survival analysis was used to find the different survival rates in the SMYD2 high expression group and low expression group. Through immunohistochemistry, the association between SMYD2 expression and clinical-pathological features of cervical cancer patients was further evaluated. Quantitative PCR and Immunoblot were applied to investigate the relative mRNA and protein expression levels, respectively. In vivo and in vitro experiments were performed to explore the function of SMYD2 in cancer progression. RESULTS: We first found a high expression of SMYD2 in cervical cancer, and survival analysis found that the poorer survival rate in the SMYD2 high expression group than that in the low expression group. Herein, our study demonstrated that the expression of SMYD2 in patients with cervical cancer was associated with FIGO stage, tumor size and further correlated with poor prognosis. Our data further showed that the inhibition of SMYD2 expression in cervical cancer cell line Caski and Siha could dramatically block the proliferation of cervical cancer cells. Additionally, SMYD2-shRNA lentivirus infected remarkably inhibited tumorigenesis in mice compared with the scramble group. CONCLUSIONS: Taken together, this study provides strong evidence of the involvement of SMYD2 in cervical cancer growth and indicates that it could have high potential as a therapeutic target of cervical cancer.
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Cervical cancer is one of the most common malignant neoplasms in gynecology. Protein tyrosine kinase 7 (PTK7) with an inactive kinase domain is an important regulator of multiple Wnt pathways under normal and various pathological conditions and overexpressed in various tumors; however, the clinical and biological significance of PTK7 in cervical cancer is still unknown. In the present study, the protein expression level of PTK7 was detected in clinical cervical cancer patient samples, and the relationship between PTK7 expression and clinicopathological features was analyzed. In addition, the Kaplan-Meier method was performed to estimate the overall survival (OS) and progression-free survival (PFS) of patients to investigate the clinicopathological significance of PTK7 expression. Functional assays demonstrated that knocking down PTK7 might inhibit the ability of cancer cells to proliferate and invade or migrate, both in vivo and in vitro. Thus, PTK7 might serve as a potential target for cervical cancer.
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Biomarcadores Tumorais/genética , Moléculas de Adesão Celular/genética , Receptores Proteína Tirosina Quinases/genética , Neoplasias do Colo do Útero/genética , Animais , Biomarcadores Tumorais/metabolismo , Moléculas de Adesão Celular/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Feminino , Humanos , Camundongos , Receptores Proteína Tirosina Quinases/metabolismo , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
The fibroblast growth factor receptors (FGFRs) are increasingly considered attractive targets for therapeutic cancer intervention due to their roles in tumor metastasis and angiogenesis. Here, we identified a new selective FGFR inhibitor, C11, and assessed its antitumor activities. C11 was a selective FGFR1 inhibitor with an IC50 of 19 nM among a panel of 20 tyrosine kinases. C11 inhibited cell proliferation in various tumors, particularly bladder cancer and breast cancer. C11 also inhibited breast cancer MDA-MB-231 cell migration and invasion via suppression of FGFR1 phosphorylation and its downstream signaling pathway. Suppression of matrix metalloproteinases 2/9 (MMP2/9) was associated with the anti-motility activity of C11. Furthermore, the anti-angiogenesis activity of C11 was verified in endothelial cells and chicken chorioallantoic membranes (CAMs). C11 inhibited the migration and tube formation of HMEC-1 endothelial cells and inhibited angiogenesis in a CAM assay. In sum, C11 is a novel selective FGFR1 inhibitor that exhibits potent activity against breast cancer metastasis and angiogenesis.
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Inibidores da Angiogênese/farmacologia , Antineoplásicos/farmacologia , Imidas/farmacologia , Perileno/análogos & derivados , Perileno/farmacologia , Receptores de Fatores de Crescimento de Fibroblastos/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Galinhas , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/fisiologia , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Metástase Neoplásica , Neovascularização Fisiológica/efeitos dos fármacos , Transdução de SinaisRESUMO
Pre-clinical diagnosis of many diseases required quantitative detection of Human Serum Albumin (HSA). Herein a high-selective HSA sensor RhHSA was picked through a two-round selectivity evolution from the typical "Effector-π-Trigger" style. RhHSA suggested advantages including high selective (â¼6 fold for HSA:BSAâ¯=â¯1:10), sensitive (LODâ¯â¼â¯5â¯nM, over 700-fold enhancement), steady (over 24â¯h) and wide linear range (0-0.5â¯mg/mL, applicative for conventional HSA measurement). The detecting system was free from media polarity or viscosity. HSA destruction, site competition and molecular docking provided reliable evidence for the fact that RhHSA could be embedded into both ibuprofen and phenylbutazone sites of HSA. These hints also supported the discrimination of HSA from BSA. Stepwisely fluid replacement in living cells and measuring in urine system both inferred the potential of RhHSA in biological applications.
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Corantes Fluorescentes/química , Soroalbumina Bovina/análise , Albumina Sérica Humana/análise , Espectrometria de Fluorescência , Animais , Sítios de Ligação , Bovinos , Corantes Fluorescentes/metabolismo , Humanos , Limite de Detecção , Células MCF-7 , Microscopia de Fluorescência , Simulação de Acoplamento Molecular , Ligação Proteica , Teoria Quântica , Rodaminas/química , Soroalbumina Bovina/metabolismo , Albumina Sérica Humana/isolamento & purificação , Albumina Sérica Humana/metabolismoRESUMO
A series of novel selective BRAFV600E inhibitory agents (Compound 1-16) 5-(2,3-dihydrobenzo[b][1,4]dioxane-6-yl)-N,3-diaryl-4,5-dihydro-1H-pyrazole-1-carbothioamides have been designed and synthesized. Their anti-proliferation and BRAF inhibitory activities were evaluated. Though 15, 4 and 12 all displayed comparable activity with the positive control Vemurafenib, only 12 indicated fine selectivity on BRAFV600E (IC50â¯=â¯0.06⯵M for BRAFV600E; GI50â¯=â¯0.52⯵M for A375) over BRAFWT at both kinase and cell levels. This result satisfied the designing concept of improving activity and introducing selectivity. Flow cytometry analysis and western blot convinced the apoptosis induction and kinase inhibitory activity. Docking simulation inferred the differences in binding patterns of BRAFV600E and BRAFWT, pointing out that the future orientation might be seeking for outer space binding of BRAFV600E and avoiding interactions with HIS573 of BRAFWT. These results brought potent BRAF inhibitors one step further to selective agents, enhancing the potential for safe medication.
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Antineoplásicos/farmacologia , Dioxanos/farmacologia , Desenho de Fármacos , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas B-raf/antagonistas & inibidores , Pirazóis/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Dioxanos/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Pirazóis/síntese química , Pirazóis/química , Relação Estrutura-AtividadeRESUMO
Four new dolabellane-type diterpene alkaloids, glandulamines Aâ-âD (1: â-â4: ), together with twelve known compounds (5: â-â16: ), were isolated from the seeds of Nigella glandulifera using repeated column chromatography and semipreparative HPLC. The structures of 1: â-â16: were elucidated based on NMR data analysis, HRMS experiments and other spectroscopic interpretations. The absolute configuration of 5: was determined by single-crystal X-ray diffraction data for the first time. Compounds 10: and 12: showed human dihydroorotate dehydrogenase inhibitory activity with IC50 values of 61.1 ± 5.3 and 45.9 ± 3.0 µM, respectively. Molecular docking of the active compound 12: and positive control teriflunomide on the inhibitor-binding site of human dihydroorotate dehydrogenase was subsequently performed to visualize the interaction pattern. In addition, compounds 8: and 10: exhibited inhibitory effects against lipopolysaccharide-induced nitric oxide production with inhibition rates of 61 and 41%, respectively, at the concentration of 10 µM. Compounds 9: and 12: showed cytotoxic activities with cell viability varying from 29 ~ 57% at 100 µM against T98G, U87, U251, and GL261 glioma cancer cell lines. These data provide new insights on the pharmacologically active compounds of this plant widely used in folk medicine.
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Alcaloides/farmacologia , Anti-Inflamatórios/farmacologia , Citotoxinas/farmacologia , Diterpenos/farmacologia , Nigella/química , Oxirredutases atuantes sobre Doadores de Grupo CH-CH/antagonistas & inibidores , Sementes/química , Alcaloides/química , Alcaloides/isolamento & purificação , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Di-Hidro-Orotato Desidrogenase , Diterpenos/química , Diterpenos/isolamento & purificação , Humanos , Técnicas In Vitro , Óxido Nítrico/antagonistas & inibidores , Difração de Raios XRESUMO
OBJECTIVE: To investigate the role of LncRNA MALAT-1 in hypoxia-induced cell injury. METHODS: Pheochromocytoma-12 (PC12) cells were divided into seven groups: Control group, Hypoxia group (Cells treated with CoCl2), MALAT-1 group (Hypoxic cells treated with MALAT-1), NC group (Hypoxic cells treated with empty plasmid), MALAT-1 siRNA group (Hypoxic cells treated with siRNA MALAT-1), SB203580 group (Hypoxic cells treated with p38MAPK inhibitor), and MALAT-1â¯+â¯SB20358 group. The content of reactive oxygen species (ROS), malondialdehyde (MDA), super oxide dismutase (SOD) and lactate dehydrogenase (LDH) was determined. Cell viability was detected by MTT assay. Apoptotic cells were observed by Hoechst 33258 and TUNEL staining assay. Mitochondrial membrane potential (MMP) was measured using JC1 vital dye. RESULTS: The decreased cell viability and increased expressions of MALAT-1 and p-p38 were observed in hypoxic PC12 cells time-dependently (Pâ¯<â¯0.05). Besides, hypoxic PC12 cells had an elevation in p-p38, ROS, MDA and LDH with the increased apoptotic cells, but a reduction in SOD and MMP, and these similar changes were more obvious in those hypoxic cells treated with MALAT-1 when compared with Controls (all Pâ¯<â¯0.05). However, the hypoxic PC12 cells treated with SB203580 and MALAT-1 siRNA led to opposite results compared with MALAT-1 group (all Pâ¯<â¯0.05). Importantly, SB203580 could reverse the function of MALAT-1 in aggravating the hypoxia injury of PC12 cells. CONCLUSION: MALAT-1 can promote the apoptosis and oxidative stress of PC12 cells by activating p38MAPK pathway, thus aggravating the damage of PC12 cells induced by chemical hypoxia.
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Hipóxia Celular/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , RNA Longo não Codificante/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cobalto/farmacologia , Malondialdeído/metabolismo , Células PC12 , Ratos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Oxidative stress and neuroinflammation are highly relevant to the pathological processes of various neurodegenerative diseases including Alzheimer's disease (AD). (+)-2-(1-hydroxyl-4-oxocyclohexyl) ethyl caffeate (HOEC), a novel 5-lipoxygenase inhibitor, was isolated from the whole plant of Incarvillea mairei var granditlora (Wehrhahn) Grierson. In this study, we investigated the protective effect of HOEC on hydrogen peroxide (H2O2) and lipopolysaccharide (LPS) -induced cytotoxicity and neuroinflammation in vitro and in vivo. MTT assay, LDH release assay, morphological observation and Hoechst 33342/PI dual staining followed by EIA, immunofluorescence staining and Western Blotting analysis were performed to elucidate the neuroprotective effect of HOEC. Treatment with HOEC at various concentrations prior to H2O2 exposure significantly enhanced cell viability, decreased LDH release, prevented cell morphologic changes and apoptosis. Instead of PGE2 reduction, HOEC markedly inhibited the production of LTB4 and suppressed the macrophage-mediated neurotoxicity. Western blotting and immunofluorescence staining showed that HOEC inhibited H2O2-induced p38 phosphorylation and NF-κB activation. Neuroprotective effect of HOEC was abolished by a p38 inhibitor. Further in vivo studies of LPS-induced neuroinflammation confirmed the anti-inflammatory effects of HOEC. These findings that HOEC protects SH-SY5Y cells from H2O2 and LPS-induced injury via arachidonic acid network modulation followed by p38 MAPK and NF-κB signaling, might make HOEC be considered as a therapeutic candidate for prevention and treatment of neurodegenerative diseases involving oxidative stress or/and inflammation.
Assuntos
Ácido Araquidônico/metabolismo , Ácidos Cafeicos/farmacologia , Cicloexanonas/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Ácidos Cafeicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cicloexanonas/química , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Humanos , Peróxido de Hidrogênio/toxicidade , Lipopolissacarídeos/toxicidade , Masculino , Camundongos Endogâmicos ICR , Neuroimunomodulação/efeitos dos fármacos , Neuroimunomodulação/fisiologia , Fármacos Neuroprotetores/química , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Distribuição AleatóriaRESUMO
The biflavonoid isochamaejasmin is mainly distributed in the root of Stellera chamaejasme L. (Thymelaeaceae) that is used in traditional Chinese medicine (TCM) to treat tumors, tuberculosis, and psoriasis. Herein, isochamaejasmin was found to show similar bioactivity against Bcl-2 family proteins to the reference Bcl-2 ligand (-)-gossypol through 3D similarity search. It selectively bound to Bcl-xl and Mcl-1 with Ki values being 1.93 ± 0.13 µmol·L(-1) and 9.98 ± 0.21 µmol·L(-1), respectively. In addition, isochamaejasmin showed slight growth inhibitory activity against HL-60 with IC50 value being 50.40 ± 1.21 µmol·L(-1) and moderate growth inhibitory activity against K562 cells with IC50 value being 24.51 ± 1.62 µmol·L(-1). Furthermore, isochamaejasmin induced apoptosis of K562 cells by increasing the intracellular expression levels of proteins of the cleavage of caspase-9, caspase-3, and PARP which involved in the Bcl-2-induced apoptosis pathway. These results indicated that isochamaejasmin induces apoptosis in leukemia cells by inhibiting the activity of Bcl-2 family proteins, providing evidence for further studying the underlying anti-cancer mechanism of S. chamaejasme L.