RESUMEN
Ulcerative colitis (UC), a pathological condition of inflammatory bowel disease, is a chronic inflammatory disorder that involves an abnormal immune response and epithelial barrier dysfunction. Although we have previously reported the anti-inflammatory effects of 7-hydroxyl-1-methylindole-3-acetonitrile (7-HMIA), a synthesized analog of arvelexin on macrophages and paw edema, its anti-colitis effect and its mechanism are not known. In this study, colitis was induced in mice model by 4% (w/v) dextran sodium sulfate (DSS) solution in drinking water for 9 days. At the same time, from the first day of administering drinking water containing DSS, the animals were treated with 5-aminosalicylic acid (5-ASA), 75 mg/kg/day, orally) or 7-HMIA (10 or 20 mg/kg/day, intraperitoneally), depending on the experimental group, respectively. The studies were terminated on the tenth day of the experiment. Our data showed that 7-HMIA reduced the disease activity index and spleen/body weight (S/B) ratio, and improved the shortened colon length comparable to the effects of 5-ASA observed in the DSS-exposed mice. 7-HMIA, like 5-ASA, inhibited the histological damage, such as a thickened colonic muscle layer and shortened crypt length in the colon of the mice with DSS-induced colitis. 7-HMIA restored the tight junction-related proteins (occludin, claudin-1, and claudin-2) and epithelial-mesenchymal transition-mediated proteins (E-cadherin, N-cadherin, and vimentin) in the colon tissue of mice with DSS-induced colitis. Additionally, 7-HMIA (20 mg/kg/day) showed the inhibitory effects similar to that of 5-ASA on the myeloperoxidase activity, interleukin (IL)-6 production, and expression levels of inducible nitric oxide synthase (iNOS), and even showed greater inhibition of IL-1ß production in the DSS-induced mice. Furthermore, the DSS-induced activation of nuclear factor-kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3) were effectively suppressed by 7-HMIA treatment like the effects of 5-ASA. Overall, our findings revealed that 7-HMIA decreased the severity of colitis by protecting the inflamed mucosal barrier by interfering with NF-κB and STAT3 activation.
Asunto(s)
Colitis Ulcerosa , Colitis , Agua Potable , Ratones , Animales , FN-kappa B/metabolismo , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Inflamación/metabolismo , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/metabolismo , Colon/patología , Mesalamina/farmacología , Mesalamina/uso terapéutico , Sulfato de Dextran/toxicidad , Modelos Animales de Enfermedad , Ratones Endogámicos C57BLRESUMEN
Due to the low bioavailability and severe toxic side effects caused by the lack of selectivity of traditional chemotherapy drugs, the targeted delivery of chemotherapy drugs has become the key to tumor treatment. The activity and transmembrane potential of mitochondria in cancer cells were significantly higher than that of normal cells, making them a potential target for chemotherapeutic drug delivery. In this study, triphenylphosphine (TPP) based mitochondria targeting polylactic acid (PLLA) nanoparticles (TPP-PLLA NPs) were synthesized to improve the delivery efficiency of anticancer drugs. The carrier material was characterized by 1H NMR and FT-IR and 7-hydroxyl coumarin (7-HC) was successfully loaded into TPP-PLLA to form 7-HC/TPP-PLLA NPs. Further studies showed that TPP-PLLA NPs were primarily accumulated in the mitochondrial and 7-HC/TPP-PLLA NPs had higher antitumor activity. Taken together, our results indicated that TPP-PLLA NPs could be a promising mitochondria-targeted drug delivery system for cancer therapy.
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Antineoplásicos , Nanopartículas , Antineoplásicos/química , Línea Celular Tumoral , Cumarinas , Portadores de Fármacos/farmacología , Sistemas de Liberación de Medicamentos , Mitocondrias , Nanopartículas/química , Compuestos Organofosforados , Poliésteres , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Cytosolic sulfotransferases (SULTs) catalyze the transfer of a sulfonate group from the cofactor 3'-phosphoadenosine 5'-phosphosulfate to a hydroxyl (OH) containing substrate and play a critical role in the homeostasis of endogenous compounds, including hormones, neurotransmitters, and bile acids. In human, SULT2A1 sulfonates the 3-OH of bile acids; however, bile acid metabolism in mouse is dependent on a 7α-OH sulfonating SULT2A8 via unknown molecular mechanisms. In this study, the crystal structure of SULT2A8 in complex with adenosine 3',5'-diphosphate and cholic acid was resolved at a resolution of 2.5 Å. Structural comparison with human SULT2A1 reveals different conformations of substrate binding loops. In addition, SULT2A8 possesses a unique substrate binding mode that positions the target 7α-OH of the bile acid close to the catalytic site. Furthermore, mapping of the critical residues by mutagenesis and enzyme activity assays further highlighted the importance of Lys44 and His48 for enzyme catalysis and Glu237 in loop 3 on substrate binding and stabilization. In addition, limited proteolysis and thermal shift assays suggested that the cofactor and substrates have protective roles in stabilizing SULT2A8 protein. Together, the findings unveil the structural basis of bile acid sulfonation targeting 7α-OH and shed light on the functional diversity of bile acid metabolism across species.
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Ácidos y Sales BiliaresRESUMEN
Veratramine, a major alkaloid from Veratrum nigrum L., has distinct anti-tumor and anti-hypertension effects. Our previous study indicated that veratramine had severe toxicity toward male rats. In order to elucidate the underling mechanism, in vivo pharmacokinetic experiments and in vitro mechanistic studies have been conducted. Veratramine was administrated to male and female rats intravenously via the jugular vein at a dose of 50 µg/kg or orally via gavage at 20 mg/kg. As a result, significant pharmacokinetic differences were observed between male and female rats after oral administration with much lower concentrations of veratramine and 7-hydroxyl-veratramine and higher concentrations of veratramine-3-O-sulfate found in the plasma and urine of female rats. The absolute bioavailability of veratramine was 0.9% in female rats and 22.5% in male rats. Further experiments of veratramine on Caco-2 cell monolayer model and in vitro incubation with GI content or rat intestinal subcellular fractions demonstrated that its efficient passive diffusion mediated absorption with minimal intestinal metabolism, suggesting no gender-related difference during its absorption process. When veratramine was incubated with male or female rat liver microsomes/cytosols, significant male-predominant formation of 7-hydroxyl-veratramine and female-predominant formation of veratramine-3-O-sulfate were observed. In conclusion, the significant gender-dependent hepatic metabolism of veratramine could be the major contributor to its gender-dependent pharmacokinetics.
Asunto(s)
Antihipertensivos/farmacocinética , Caracteres Sexuales , Alcaloides de Veratrum/farmacocinética , Administración Intravenosa , Administración Oral , Animales , Antihipertensivos/administración & dosificación , Células CACO-2 , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Ratas , Ratas Sprague-Dawley , Distribución Tisular/efectos de los fármacos , Distribución Tisular/fisiología , Alcaloides de Veratrum/administración & dosificaciónRESUMEN
Objective: To study the flavonoids constituents from the thorns of Gleditsia sinensis and their cytotoxicity against tumor cells. Methods: The compounds were isolated by silica gel, Sephadex LH-20 column chromatography, and HPLC techniques. Their structures were elucidated on the basis of spectroscopic analyses. Results: Twelve flavonoids were obtained and identified as (2R,3R)-5,3',4'-trimethoxyl-7-hydroxyl-flavanonol (1), 5,7,3',4'-tetrahydroxyl-flavanonol (2), 5-methoxyl-3',4',7-trihydroxyl-flavanonol (3), dihydrokaempferol (4), epicatechin (5), 5,7,3',5'-tetrahydroxyl-flavanonol (6), fustin (7), (2R,3R)-7,3',5'-trihydroxyl-flavanonol (8), (2R,3R)-5,7,3'-trihydroxyl-4'-methoxyl-flavanonol (9), quercetin (10), 5,7,4'-trihydroxylflavone-8-C-glucopyranose (11), and 2,7-dimethyl-xanthone (12), respectively. Conclusion: Compound 1 is a new component named G. spina flavanonol A, and compounds 3,8,9, and 12 are isolated from the thorns of G. sinensis for the first time. The results of cytotoxicity test show that the dihydroflavonol compound 7 displays the stronger cytotoxicity against HepG2, A549, and EC109 cell strains, while compounds 1 and 3 have the effects on HepG2 and EC109, and compound 2 has the effect on EC109 cancer cells, respectively.
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Aim To investigate the effects of 1 7-me-thoxyl-7-hydroxyl-benzofuran chalcone(YLSC)on my-ocardial ischemia /reperfusion injury(MI /RI)by mod-ulating PI3K/Akt signaling pathway and the possible mechanisms.Methods Male SD rats were randomly divided into sham group,model group,YLSC group, wortmannin(WM)group and YLSC +WM group(n =8).The rat model of MI /RI was established by ligation of the left anterior descending artery for 30 min fol-lowed by loosening the ligature for 2 h.After reperfu- sion,blood samples were obtained to determine serum contents of CK-MB,LDH,NO and TNF-α.The pro-tein levels of total (t)-Akt,phosphorylated (p)-Akt and LC3-Ⅱ were detected by Western blot.Caspase-3,Beclin1 and eNOS mRNA expression was evaluated by FQ-PCR.Results YLSC pretreatment greatly re-duced serum levels of CK-MB,LDH and TNF-α,and elevated NO content.It also inhibited the expression of caspase-3,Beclin1 and LC3-Ⅱ,while enhanced p-Akt and eNOS expression.Conclusion YLSC protects the heart against MI /RI via inhibition of apoptosis and excessive autophagy,in which protective effect is regu-lated by activation of the PI3K/Akt pathway.
RESUMEN
We previously demonstrated that 7-hydroxyl-1-methylindole-3-acetonitrile (7-HMIA), a synthesized analog of arvelexin, showed the strong inhibitory effects on LPS-induced NO and PGE2 production in macrophages. In this study, we focused on elucidating the anti-inflammatory properties of 7-HMIA and the mechanisms involved using in vitro and in vivo experimental models. In LPS-induced RAW 264.7 macrophages, 7-HMIA significantly inhibited the release of proinflammatory mediators such as prostaglandin E2 (PGE2), nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). We also found that 7-HMIA suppressed PGE2 production not by inhibiting cyclooxygenase-2 (COX-2) expression or activity, but rather by suppressing the mRNA stability of microsomal prostaglandin E synthase (mPGES-1). Furthermore, 7-HMIA mediated attenuation of inducible NO synthase (iNOS), TNF-α, and IL-6 was closely associated with suppression of transcriptional activities of nuclear factor-kappa B (NF-κB), by decreasing p65 nuclear translocation and Akt phosphorylation. Animal studies revealed that 7-HMIA potently suppressed the carrageenan-induced paw edema and myeloperoxidase (MPO) activity in paw tissues. Taken together, our data indicated that the molecular basis for the anti-inflammatory properties of 7-HMIA involved the inhibition of mRNA stability of mPGES-1 and PI3K/Akt-mediated NF-κB pathways.