RESUMEN
The folate metabolism enzyme ALDH1L1 catalyzed 10-formyltetrahydrofolate to tetrahydrofolate and CO2. Non-small cell lung cancer cells (NSCLC) strongly express ALDH1L1. Gossypol binds to an allosteric site and disrupts the folate metabolism by preventing NADP+ binding. The Cryo-EM structures of tetrameric C-terminal aldehyde dehydrogenase human ALDH1L1 complex with gossypol were examined. Gossypol-bound ALDH1L1 interfered with NADP+ by shifting the allosteric site of the structural conformation, producing a closed-form NADP+ binding site. In addition, the inhibition activity of ALDH1L1 was targeted with gossypol in NSCLC. The gossypol treatment had anti-cancer effects on NSCLC by blocking NADPH and ATP production. These findings emphasize the structure characterizing ALDH1L1 with gossypol.
Asunto(s)
Gosipol , Humanos , Gosipol/química , Gosipol/farmacología , Gosipol/metabolismo , NADP/metabolismo , NADP/química , Modelos Moleculares , Microscopía por Crioelectrón , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/patología , Aldehído Oxidorreductasas/metabolismo , Aldehído Oxidorreductasas/química , Unión Proteica , Sitios de Unión , Sitio Alostérico , Conformación Proteica , Línea Celular Tumoral , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NHRESUMEN
ATP2B1 is a known regulator of calcium (Ca2+) cellular export and homeostasis. Diminished levels of intracellular Ca2+ content have been suggested to impair SARS-CoV-2 replication. Here, we demonstrate that a nontoxic caloxin-derivative compound (PI-7) reduces intracellular Ca2+ levels and impairs SARS-CoV-2 infection. Furthermore, a rare homozygous intronic variant of ATP2B1 is shown to be associated with the severity of COVID-19. The mechanism of action during SARS-CoV-2 infection involves the PI3K/Akt signaling pathway activation, inactivation of FOXO3 transcription factor function, and subsequent transcriptional inhibition of the membrane and reticulum Ca2+ pumps ATP2B1 and ATP2A1, respectively. The pharmacological action of compound PI-7 on sustaining both ATP2B1 and ATP2A1 expression reduces the intracellular cytoplasmic Ca2+ pool and thus negatively influences SARS-CoV-2 replication and propagation. As compound PI-7 lacks toxicity in vitro, its prophylactic use as a therapeutic agent against COVID-19 is envisioned here.
Asunto(s)
COVID-19 , Calcio , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , SARS-CoV-2 , Transducción de Señal , Replicación Viral , Humanos , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/fisiología , Replicación Viral/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , COVID-19/virología , COVID-19/metabolismo , Transducción de Señal/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Calcio/metabolismo , Animales , Proteína Forkhead Box O3/metabolismo , Proteína Forkhead Box O3/genética , Chlorocebus aethiops , Tratamiento Farmacológico de COVID-19 , Células Vero , Femenino , ATPasas Transportadoras de Calcio/metabolismo , ATPasas Transportadoras de Calcio/genética , MasculinoRESUMEN
Inorganic polyphosphates (polyPs) are linear polymers composed of repeated phosphate (PO4 3-) units linked together by multiple high-energy phosphoanhydride bonds. In addition to being a source of energy, polyPs have cytoprotective and antiviral activities. Here, we investigated the antiviral activities of long-chain polyPs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In molecular docking analyses, polyPs interacted with several conserved amino acid residues in angiotensin-converting enzyme 2 (ACE2), the host receptor that facilitates virus entry, and in viral RNA-dependent RNA polymerase (RdRp). ELISA and limited proteolysis assays using nano- LC-MS/MS mapped polyP120 binding to ACE2, and site-directed mutagenesis confirmed interactions between ACE2 and SARS-CoV-2 RdRp and identified the specific amino acid residues involved. PolyP120 enhanced the proteasomal degradation of both ACE2 and RdRp, thus impairing replication of the British B.1.1.7 SARS-CoV-2 variant. We thus tested polyPs for functional interactions with the virus in SARS-CoV-2-infected Vero E6 and Caco2 cells and in primary human nasal epithelial cells. Delivery of a nebulized form of polyP120 reduced the amounts of viral positive-sense genomic and subgenomic RNAs, of RNA transcripts encoding proinflammatory cytokines, and of viral structural proteins, thereby presenting SARS-CoV-2 infection in cells in vitro.
Asunto(s)
Antivirales/farmacología , Tratamiento Farmacológico de COVID-19 , Polifosfatos/farmacología , SARS-CoV-2/efectos de los fármacos , Administración por Inhalación , Secuencia de Aminoácidos , Enzima Convertidora de Angiotensina 2/química , Enzima Convertidora de Angiotensina 2/metabolismo , Animales , Antivirales/administración & dosificación , Antivirales/química , COVID-19/metabolismo , COVID-19/virología , Células CACO-2 , Chlorocebus aethiops , ARN Polimerasa Dependiente de ARN de Coronavirus/química , ARN Polimerasa Dependiente de ARN de Coronavirus/genética , ARN Polimerasa Dependiente de ARN de Coronavirus/metabolismo , Citocinas/metabolismo , Células HEK293 , Interacciones Microbiota-Huesped/efectos de los fármacos , Interacciones Microbiota-Huesped/genética , Interacciones Microbiota-Huesped/fisiología , Humanos , Técnicas In Vitro , Modelos Biológicos , Simulación del Acoplamiento Molecular , Nebulizadores y Vaporizadores , Polifosfatos/administración & dosificación , Polifosfatos/química , Complejo de la Endopetidasa Proteasomal/metabolismo , Dominios y Motivos de Interacción de Proteínas , Proteolisis/efectos de los fármacos , ARN Viral/genética , ARN Viral/metabolismo , SARS-CoV-2/genética , SARS-CoV-2/fisiología , Homología de Secuencia de Aminoácido , Transducción de Señal/efectos de los fármacos , Células Vero , Replicación Viral/efectos de los fármacosRESUMEN
Red ginseng is one of the most popular traditional medicines in Korea because its soluble hot-water extract is known to be very effective on enhancing immunity as well as inhibiting inflammation. Recently, we developed a new technique, called the HAC-gearshift system, which can pulverize red ginseng into the ultra-fine granules ranging from 0.2 to 7.0 microm in size. In this study, the soluble hot-water extract of those ultra-fine granules of red ginseng (URG) was investigated and compared to that of the normal-sized granules of red ginseng (RG). The high pressure liquid chromatographic analyses of the soluble hot-water extracts of both URG and RG revealed that URG had about 2-fold higher amounts of the ginsenosides, the biologically active components in red ginseng, than RG did. Using quantitative RT-PCR, cytokine profiling against the Escherichia coli lipopolysaccharide (LPS) in the monocyte-derived macrophage THP-1 cells demonstrated that the URG-treated cells showed a significant reduction in cytokine expression than the RG-treated ones. Transcription expression of the LPS-induced cytokines such as TNF-alpha, IL-1beta, IL-6, IL-8, IL-10, and TGF-beta was significantly inhibited by URG compared to RG. These results suggest that some biologically active and soluble components in red ginseng can be more effectively extracted from URG than RG by standard hot-water extraction.
RESUMEN
BR003 is a multi-herbal formula that contains twelve medicinal herbs. We investigated the effects of oral administration of BR003 to Wistar rats on (a) learning and memory using a passive avoidance test and (b) cell proliferation in the dentate gyrus (DG) of the hippocampus using immunohistochemical analysis of 5-bromo-2-deoxyuridine (BrdU) expression. In the passive avoidance test, the retention time of the BR003-treated group was significantly longer than that of the control group (182.64+/-39.88 vs. 73.08+/-29.30 s, respectively; n=11; p<0.05). There were significantly more BrdU-immunoreactive cells in the DG in the BR003-treated group than in the control group (1281.07+/-151.16 vs. 818.01+/-132.98 cells per DG, respectively; n=11; p<0.05). These results suggest that the administration of BR003 not only improves learning and memory but also increases cell proliferation in the DG of the rat hippocampus.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Giro Dentado/citología , Giro Dentado/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Memoria/efectos de los fármacos , Nootrópicos/farmacología , Animales , Antimetabolitos/farmacología , Reacción de Prevención/efectos de los fármacos , Bromodesoxiuridina/farmacología , Inmunohistoquímica , Masculino , Ratas , Ratas Wistar , Estimulación QuímicaRESUMEN
Moutan Cortex Radicis (MCR) is one of the most widely used Oriental medicines. In this study, we assessed the reducing effect of ethanol extract of MCR on hydrogen peroxide-induced reactive oxygen production, the main cause of cell damage or death in PC12 cells. The viability of cells treated with 1 mg/ml of MCR was significantly restored from that of oxidative-stressed PC12 cells. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H(2)DCFDA assay. MCR at 1-0.01 mg/ml concentration inhibited ROS production in oxidative-stressed cells. To identify candidate genes responsible for the anti-oxidative effects of MCR on PC12 cells, an oligonucleotide microarray analysis was performed. The result of gene expression profiles showed that 10 genes were up-regulated and 7 were down-regulated in MCR plus hydrogen peroxide treated cells compared with hydrogen peroxide treated cells. Among them, heme oxygenase (HO) and cathechol-O-methyltransferase (COMT) are related to regulation of ROS generation and the others are known to regulate cell survival and progression. Subsequently, we performed real-time RT-PCR to quantify the ROS related gene. MCR treatment increased the expression of HO by 370% and COMT by 280% at the concentration of 1 mg/ml. These findings suggest that MCR inhibits the production of ROS and cytotoxicity by oxidative-stressed PC12 cells through over-expression of HO and COMT.
Asunto(s)
Perfilación de la Expresión Génica , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/metabolismo , Animales , Regulación hacia Abajo , Sistemas Neurosecretores/efectos de los fármacos , Estrés Oxidativo , Células PC12 , Paeonia , Ratas , Regulación hacia ArribaRESUMEN
AIM: Taraxacum officinale (TO) has been frequently used as a remedy for inflammatory diseases. The aim of this study was to investigate the effect of TO on cholecystokinin (CCK)-octapeptide-induced acute pancreatitis in rats. METHODS: TO at 10 mg/kg was orally administered, followed by 75 microg/kg CCK octapeptide injected subcutaneously three times after 1, 3 and 5 h. This whole procedure was repeated for 5 d. We determined the pancreatic weight/body weight ratio, the levels of pancreatic HSP60 and HSP72, and the secretion of pro-inflammatory cytokines. Repeated CCK octapeptide treatment resulted in typical laboratory and morphological changes of experimentally-induced pancreatitis. RESULTS: TO significantly decreased the pancreatic weight/body weight ratio in CCK octapeptide-induced acute pancreatitis. TO also increased the pancreatic levels of HSP60 and HSP72. Additionally, the secretion of IL-6 and TNF-alpha decreased in the animals treated with TO. CONCLUSION: TO may have a protective effect against CCK octapeptide-induced acute pancreatitis.