RESUMEN
BACKGROUND: Despite the notable pharmacological potential of natural ginsenosides, their industrial application is hindered by low oral bioavailability. Recent research centers on the production of less-glycosylated minor ginsenosides. PURPOSE: This study aimed to explore the effect of a biologically synthesized ginsenoside CK-rich minor ginsenoside complex (AceCK40), on ameliorating colitis using DSS-induced colitis models in vitro and in vivo. METHODS: The ginsenoside composition of AceCK40 was determined by HPLC-ELSD and UHPLC-MS/MS analyses. In vitro colitis model was established using dextran sodium sulfate (DSS)-induced Caco-2 intestinal epithelial model. For in vivo experiments, DSS-induced severe colitis mouse model was established. RESULTS: In DSS-stimulated Caco-2 cells, AceCK40 downregulated mitogen-activated protein kinase (MAPK) activation (p < 0.05), inhibited monocyte chemoattractant protein-1 (MCP-1) production (p < 0.05), and enhanced MUC2 expression (p < 0.05), mediated via signaling pathway regulation. Daily AceCK40 administration at doses of 10 and 30 mg/kg/day was well tolerated by DSS-induced severe colitis mice. These doses led to significant alleviation of disease activity index score (> 36.0% decrease, p < 0.05), increased luminal immunoglobulin (Ig)G (> 37.6% increase, p < 0.001) and IgA (> 33.8% increase, p < 0.001), lowered interleukin (IL)-6 (> 65.7% decrease, p < 0.01) and MCP-1 (> 116.2% decrease, p < 0.05), as well as elevated serum IgA (> 51.4% increase, p < 0.001) and lowered serum IL-6 (112.3% decrease at 30 mg/kg, p < 0.001). Hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining revealed that DSS-mediated thickening of the muscular externa, extensive submucosal edema, crypt distortion, and decreased mucin droplets were significantly alleviated by AceCK40 administration. Additionally, daily administration of AceCK40 led to significant recovery of colonic tight junctions damaged by DSS through the elevation in the expression of adhesion molecules, including occludin, E-cadherin, and N-cadherin. CONCLUSION: This study presents the initial evidence elucidating the anti-colitis effects of AceCK40 and its underlying mechanism of action through sequential in vitro and in vivo systems employing DSS stimulation. Our findings provide valuable fundamental data for the utilization of AceCK40 in the development of novel anti-colitis candidates.
Asunto(s)
Colitis , Ginsenósidos , Humanos , Ratones , Animales , Ginsenósidos/metabolismo , Células CACO-2 , Ratones Endogámicos C57BL , Espectrometría de Masas en Tándem , Colitis/inducido químicamente , Colitis/tratamiento farmacológico , Colitis/metabolismo , Colon , Inmunoglobulina A/metabolismo , Inmunoglobulina A/farmacología , Inmunoglobulina A/uso terapéutico , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Mucosa Intestinal/metabolismoRESUMEN
A novel, strictly anaerobic, moderately thermophilic, endospore-forming, sulfate-reducing bacterium, designated TGB60-1T, was isolated from a hydrothermal sediment vent collected from the Tofua Arc in the Tonga Trench. The strain was characterized phenotypically and phylogenetically. The isolated strain was observed to be Gram-positive, with slightly curved rod-shaped cells and a polar flagellum. Strain TGB60-1T was found to grow anaerobically at 3760 °C (optimum, 50 °C), at pH 6.08.5 (optimum, pH 7.0) and with 1.04.0 % (w/v) NaCl (optimum, 3.0 %). The electron acceptors utilised were determined to be sulfate, sulfite, and thiosulfate. Strain TGB60-1T was found to utilise pyruvate and H2 as electron donors. Strain TGB60-1T was determined to be related to representatives of the genus Desulfotomaculum and the closest relatives within this genus were identified as Desulfotomaculum halophilum SEBR 3139T, Desulfotomaculum alkaliphilum S1T and Desulfotomaculum peckii LINDBHT1T (92.7, 92.1, and 91.8 % 16S rRNA gene sequence similarity, respectively). The major fatty acids (>20 %) were identified as C16:0 and C18:1 ω7c. The G+C content of the genomic DNA of this novel bacterium was determined to be 53.9 mol%. Based on this polyphasic taxonomic study, strain TGB60-1T is considered to represent a novel species in the genus Desulfotomaculum, for which the name Desulfotomaculum tongense sp. nov. is proposed. The type strain of D. tongense is strain TGB60-1T (= KTCT 4534T = JCM 18733T).
Asunto(s)
Desulfotomaculum/clasificación , Desulfotomaculum/aislamiento & purificación , Sedimentos Geológicos/microbiología , Aerobiosis , Técnicas de Tipificación Bacteriana , Composición de Base , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Desulfotomaculum/genética , Desulfotomaculum/fisiología , Ácidos Grasos/análisis , Flagelos/fisiología , Hidrógeno/metabolismo , Concentración de Iones de Hidrógeno , Respiraderos Hidrotermales , Microscopía Electrónica , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , Ácido Pirúvico/metabolismo , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Cloruro de Sodio/metabolismo , Compuestos de Azufre/metabolismo , Temperatura , TongaRESUMEN
A frequently used fermentation manner in lactic acid (LA) production, batch fermentation by pure cultures, has a limited practicability: low volumetric productivity and high energy consumption. In this study, continuous LA fermentation was performed in a completely stirred tank reactor at 12h HRT, inoculated with anaerobic digester sludge. Glucose (25 g COD/L) was used as a feedstock and temperature was increased from 35 to 60°C. LA production significantly increased from 50°C, which was negligible up to 45°C, with obvious bacterial community change. At 50 and 55°C, LA production was maximized, reaching 23 g COD/L, corresponding to 92% LA conversion efficiency. Pyrosequencing analysis showed that microbial diversity was simplified at 50-60°C, and the sequences closely related with Bacillus coagulans became predominant, followed by Lactobacillus fermentum. An LA-producing upflow ananerobic sludge blanket reactor was successfully developed, which enhanced the productivity up to 4.8 gLA/L/h by shortening HRT to 4h.