RESUMEN
One of the important monitoring indicators of the air pollution is atmospheric fine particulate matter (PM2.5 ), which can induce lung inflammation after inhalation. Coelonin can alleviate PM2.5 -induced macrophage damage through anti-inflammation. However, its molecular mechanism remains unclear. We hypothesized that macrophage damage may involve the release of inflammatory cytokines, activation of inflammatory pathways, and pyrosis induced by inflammasome. In this study, we evaluated the anti-inflammation activity of coelonin in PM2.5 -induced macrophage and its mechanism of action. Nitric oxide (NO) and reactive oxygen species (ROS) production were measured by NO Assay kit and dichlorofluorescein-diacetate (DCFH-DA), and apoptosis were measured by Flow cytometry and TUNEL staining. The concentration of inflammatory cytokines production was measured with cytometric bead arrays and ELISA kits. The activation of NF-κB signaling pathway and NLRP3 inflammasome were measured by immunofluorescence, quantitative reverse transcription-polymerase chain reaction and western blot. As expected, coelonin pretreatment reduced NO production significantly as well as alleviated cell damage by decreasing ROS and apoptosis. It decreased generation of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in PM2.5 -induced RAW264.7 and J774A.1 cells. Moreover, coelonin markedly inhibited upregulating the expression of toll-like receptor (TLR)4 and cyclo-oxygenase (COX)-2, blocked activation of p-nuclear factor-kappa B (NF-κB) signaling pathway, and suppressed expression of NLRP3 inflammasome, ASC, GSDMD, IL-18 and IL-1ß. In conclusion, the results showed that coelonin could protect against PM2.5 -induced macrophage damage via suppressing TLR4/NF-κB/COX-2 signaling pathway and NLRP3 inflammasome activation in vitro.
Asunto(s)
Inflamasomas , FN-kappa B , FN-kappa B/metabolismo , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ciclooxigenasa 2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptor Toll-Like 4/metabolismo , Transducción de Señal , Macrófagos/metabolismo , Citocinas/metabolismo , Interleucina-6 , Antiinflamatorios/farmacología , Material Particulado/toxicidadRESUMEN
Biocontrol fungi are widely used to promote plant growth and pest control. Four fungi were isolated from Cremastra appendiculata tubers and screened for plant growth-promoting and antagonistic effects. Based on the morphological characterization and ITS, 18S rRNA and 28S rRNA gene sequencing analysis, the fungi were identified to be related to Colletotrichum gloeosporioides (DJL-6), Trichoderma tomentosum (DJL-9), Colletotrichum godetiae (DJL-10) and Talaromyces amestolkiae (DJL-15). The growth inhibition tests showed that the four isolates had different inhibitory effects on Colletotrichum fructicola, Alternaria alternata and Alternaria longipes, among which DJL-9 showed the highest inhibitory activity. Their culture filtrates (especially that of DJL-15) can also inhibit pathogens. Four isolates were positive for the production of indole-3-acid (IAA) and ß-1,3-glucanase and possessed proteolytic activity but were negative for the production of iron siderophore complexes. The four fungi showed strong nitrogen fixation and potassium dissolution abilities. In addition to DJL-9 being able to solubilize phosphate, DJL-10 was able to produce chitinase and cellulase. Pot experiments indicated that the four fungi increased the germination rate of C. appendiculata and soybean seeds and increased soybean radicle growth and plant biomass. Among them, DJL-6 had a better growth-promoting effect. Therefore, we successfully screened the biocontrol potential of endophytes from C. appendiculata, with a focus on preventing fungal diseases and promoting plant growth, and selected strains that could provide nutrients and hormones for plant growth.
Asunto(s)
Hongos , Desarrollo de la Planta , Endófitos , Fosfatos , Semillas , Enfermedades de las Plantas/microbiologíaRESUMEN
Tetrastigma hemsleyanum Diels et Gilg is a traditional Chinese herbal medicine with high medicinal value, and antitumor, antioxidant and anti-inflammatory biological activities. However, while several studies have focused on flavonoids in Tetrastigma hemsleyanum tubers, there are few studies on the enhanced immune effect of Tetrastigma hemsleyanum polysaccharides (THP). In this study, we evaluated the antitumor effect of THP in a lung tumor model and explored the mechanism of antitumor activity through intestinal flora. In addition, a cyclophosphamide (CTX)-induced immunosuppression model was used to declare the immunomodulatory effect of THP in the immunosuppressive state induced by antitumor drugs. The results showed that THP increased the content of ileum secreted immunoglobulin A (SIgA) and cecum short-chain fatty acids (SCFAs) and improved microbial community diversity, regulating the relative abundance of dominant microbiota flora from the phylum level to the genus level, and recovering the intestinal microflora disorder caused by tumors. Additionally, THP can increase the organ indices and improve immune organ atrophy. THP can upregulate routine blood counts and stimulate the production of the serum cytokines. THP also promoted the macrophage phagocytic index, NK-cell activation, and complement and immunoglobulin (IgG, IgA, IgM) levels. The detection of Splenic lymphocyte proliferation and T lymphocyte subsets also sideways reflects that THP can restore CTX-induced immune inhibition in mice. In conclusion, this study suggests that THP can effectively achieve the enhanced antitumor effects, regulate gut microbiota and improve the immunosuppression induced by antitumor drugs. Therefore, THP can enhance the immune capacity and provide novel immunomodulatory and antineoplastic adjuvant agents.
Asunto(s)
Microbioma Gastrointestinal , Vitaceae , Ratones , Animales , Polisacáridos/farmacología , Polisacáridos/uso terapéutico , Inmunidad , InmunomodulaciónRESUMEN
Tetrastigma Hemsleyanum Diels et Gilg (Sanyeqing, SYQ), a traditional herb native to China, has been widely used in the treatment of inflammatory diseases, febrile convulsions and dysentery. Previous studies have demonstrated the hypoglycemic, anti-inflammatory and anti-pyretic effects of polysaccharides from SYQ (SYQP). However, the role of SYQP in acute lung injury (ALI) remained unknown. Our present study aimed to explore the protective effects of SYQP in ALI and explained its underlying mechanism. In vivo, Balb/c mice were intragastrically administered with SYQP and dexamethasone (DXMS) for 14 days which were intratracheally instilled (IT) with LPS at the last day. In vitro, A549 cells were pretreated with SYQP, DXMS or TAK-242, following by LPS stimulation for 24 h. The histopathological results showed that SYQP remarkably alleviated pulmonary macrophage infiltration and myeloperoxidase (MPO) levels. Additionally, SYQP evidently suppressed wet/dry (W/D) ratio of lung and white blood cells (WBC) in bronchoalveolar lavage fluid (BALF). Moreover, SYQP markedly reduced TNF-α, IL-6, IL-1ß and COX-2 levels both in vivo and in vitro. Meanwhile, SYQP significantly attenuated oxidative stress through upregulating the levels of SOD and GSH-Px and downregulating the expressions of MDA and LDH in vivo, which was confirmed by in vitro results that SYQP could increase SOD activity and decrease MDA and NO contents. Importantly, a decline in protein expressions of TLR4, COX-2, NF-κB p50, phospho-NF-κB p65 and phospho-IκB-α was detected in vivo and in vitro. Taken together, our results demonstrated that SYQP ameliorated LPS-induced ALI by alleviating inflammation and oxidative stress via attenuating TLR4/COX-2/NF-κB signaling pathway.
Asunto(s)
Lesión Pulmonar Aguda , FN-kappa B , Ratones , Animales , FN-kappa B/metabolismo , Receptor Toll-Like 4/metabolismo , Lipopolisacáridos/farmacología , Peroxidasa/metabolismo , Ciclooxigenasa 2/metabolismo , Inhibidor NF-kappaB alfa/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Interleucina-6/farmacología , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/tratamiento farmacológico , Lesión Pulmonar Aguda/metabolismo , Transducción de Señal , Antiinflamatorios/efectos adversos , Superóxido Dismutasa/metabolismo , Dexametasona/farmacología , Hipoglucemiantes/farmacologíaRESUMEN
Mounting evidence supports that long-term exposure to fine particle pollutants (PM2.5) is closely implicated in cardiovascular diseases, especially atherosclerosis. Amygdalin is reported to attenuate external stimuli-induced cardiovascular diseases. However, the underlying mechanisms are still not understood. In this study, we aim to explore the protective effects of amygdalin on PM2.5-induced human umbilical vein endothelial cell (HUVEC) injury and unravel the specific mechanisms by MTT, DCFH-DA, biochemical, immunofluorescence, ELISA, RT-qPCR, flow cytometry, TUNEL and western blot analysis. The results reveal that amygdalin reverses PM2.5-induced cytotoxicity and attenuates intracellular ROS production. Moreover, amygdalin increases the levels of SOD and GSH and alleviates the MDA content. Additionally, amygdalin causes a decline of IL-6, IL-1ß, TNF-α and COX-2 levels. Moreover, amygdalin inhibits NF-κB p50 and TLR4 protein expressions and NF-κB p65 nuclear translocation. Concomitantly, a decline of phospho-NF-κB p65/NF-κB p65 and phospho-IκB-α/IκB-α is detected. Meanwhile, amygdalin pretreatment reduces HUVEC apoptosis. In addition, amygdalin triggers an upregulation of Bcl-2 and a downregulation of Bax after stimulation with PM2.5. Collectively, these results suggest that amygdalin suppresses PM2.5-induced HUVEC injury by regulating the TLR4/NF-κB and Bcl-2/Bax signaling pathways, indicating that amygdalin may be a novel target for atherosclerosis treatments.
Asunto(s)
Amigdalina , Enfermedades Cardiovasculares , Humanos , FN-kappa B/metabolismo , Amigdalina/farmacología , Proteína X Asociada a bcl-2 , Inhibidor NF-kappaB alfa/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Receptor Toll-Like 4 , Transducción de Señal , Material Particulado/toxicidadRESUMEN
BACKGROUND: Inflammation is the pathogenesis of various chronic diseases plaguing clinic for years.Fallopia multiflora (Thunb.) is a traditional Chinese herbal medicine with a long history of application in detoxification and anti-inflammation. 2,3,4',5-Tetrahydroxystilbene 2-o-D-glucoside (TSG) is a main active compound of F. multiflora. However, the mechanism of TSG in the treatment of inflammation remains unknown. METHODS: Network pharmacology and molecular docking were employed to explore the mechanism of anti-inflammatory effect of TSG. Potential targets of TSG and inflammation were obtained from Swiss Target Prediction, Pharm Mapper, and GeneCards database. Protein-protein interaction (PPI) networks, GO and KEGG pathway enrichment analysis were performed to elucidate the interaction of targets. Moreover, the anti-inflammatory effect of TSG was validated by in vitro experiments using flow cytometry, RT-qPCR, Western blot, and immunocytochemistry assays. RESULTS: PPI network and gene enrichment analysis showed that TSG may exert a protein kinase binding activity, and IKBKB, MAPK1, NFKBIA, and RELA were predicted as the targets of anti-inflammation. Verified by molecular docking and Western blot, TSG may target NF-κB and ERK2 related signals to alleviate inflammatory damage. Furthermore, TSG effectively downregulated the expression of inflammatory cytokine, the nuclear translocation of NF-κB p65, and the production of reactive oxygen species (ROS). CONCLUSION: TSG possesses significant anti-inflammatory effect. TSG may display a protein kinase binding activity and target NF-κB and ERK2 related signals to treat the inflammation. This work may enlighten the potential application of TSG in anti-inflammation and indicate network pharmacology was an effective tool for the further study of TCM.
Asunto(s)
Medicamentos Herbarios Chinos , FN-kappa B , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Medicamentos Herbarios Chinos/farmacología , Glucósidos/farmacología , Glucósidos/uso terapéutico , Humanos , Inflamación/tratamiento farmacológico , Simulación del Acoplamiento Molecular , FN-kappa B/metabolismo , Farmacología en Red , Proteínas Quinasas/uso terapéuticoRESUMEN
The optimization of extraction of Tetrastigma hemsleyanum Diels et Gilg polysaccharides (THP) using ultrasonic with enzyme method and its monosaccharide compositions and antioxidant activity were investigated in this work. Single-factor experiments and response surface methodology (RSM) were performed to optimize conditions for extraction, and the independent variables were (XA) dosage of cellulase, (XB) extraction time, (XC) ultrasonic power, and (XD) ratio of water to the material. The extraction rate of THP was increased effectively under the optimum conditions, and the maximum (4.692 ± 0.059%) was well-matched the predicted value from RSM. THP was consisted of mannose, glucuronic acid, rhamnose, galacturonic acid, glucose, galactose, and arabinose, while glucose was the dominant (26.749 ± 0.634%). According to the total antioxidant capacity assay with the FRAP method, DPPH, and hydroxyl radical scavenging assay, THP showed strong antioxidant activity with a dose-dependent behavior. The results indicated that THP has the potential to be a novel antioxidant and could expand its application in food and medicine.
Asunto(s)
Antioxidantes , Vitaceae , Antioxidantes/química , Glucosa , Monosacáridos , Polisacáridos/química , Vitaceae/químicaRESUMEN
Accumulating studies suggest that fine particulate matter (PM2.5) pollutants in the air are easily enter into alveoli and even the bloodstream, resulting in an inflammatory response that not only triggers respiratory disorders but also causes permanent damage to various organs. Recent findings suggest that coelonin and militarine enriched in orchids can inhibit inflammation-induced injury against respiratory diseases. Here, we evaluated the anti-inflammatory properties of coelonin and militarine and examined their underlying molecular mechanisms in A549 cells exposed to PM2.5. PM2.5 induced significant intracellular reactive oxidative stress accumulation at a concentration of 250 µg/ml, as determined using the dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence assay. Cell viability was assessed via the MTS assay to determine the concentrations of compounds appropriate for use in subsequent experiments. Data from the enzyme-linked immunosorbent assay (ELISA) showed that both coelonin (10 and 20 µg/ml) and militarine (5 and 10 µg/ml) mitigated PM2.5-induced inflammation by reducing the generation of inflammatory factors, including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Quantitative real-time PCR (qRT-PCR) analysis revealed a remarkable decrease in IL-6, TNF-α, cyclooxygenase-2 (COX-2) and interleukin-1ß (IL-1ß) mRNA levels in the coelonin and militarine-pretreated groups. In Western blot analysis, expression of inhibitor of NF-κB (IκBα) protein in the coelonin and militarine pretreatment groups was significantly increased compared with the PM2.5 (only) treatment group (P < 0.05), concomitant with a significant decrease in phospho-IκB kinase ß/IκB kinase ß (p-IKKß/IKKß), phospho-nuclear factor of kappa B p65/nuclear factor of kappa B p65 (p-NF-κBp65/NF-κBp65) and COX-2 proteins (P < 0.05). Both coelonin and militarine inhibited migration and inflammation by suppressing PM2.5-induced IKK phosphorylation, and followed by IκBα protein degradation and NF-κB activation. Our collective data strongly supported the utility of coelonin and militarine as novel sources for development of treatments for PM2.5-induced lung diseases.