RESUMEN
Previous reports have shown that plant-derived microRNAs (miRNAs) regulate mammalian gene expression through dietary intake. Our prior study found that gma-miR159a, which is abundant in soybean, significantly inhibited the proliferation of colon cancer cells. In the current study, dietary gma-miR159a was utilized to study its anti-colon cancer function in azoxymethane (AOM)/dextran sodium sulfate (DSS)-induced colon cancer mice. Under processing conditions, gma-miR159a exhibited excellent stability in cooked soybean. In vitro, gma-miR159a suppressed the expression of the oncogene MYC downstream of the Wnt signaling pathway by targeting the TCF7 gene, significantly inhibiting the growth of colon cancer cells. The in vivo experiments showed that gma-miR159a and soybean RNA (total RNA extracted from soybean) significantly reduced tumor growth in AOM/DSS-induced colon cancer mice by gavage. This effect disappeared when anti-miR159a was present. In addition, gma-miR159a and soybean RNA significantly attenuated inflammation in colon cancer mice. These results showed that long-term dietary intake of soybean-derived gma-miR159a effectively prevented the occurrence of colon cancer and colitis, which provides novel evidence for the prevention function of soybean.
Asunto(s)
Neoplasias del Colon/terapia , Glycine max/genética , Factor Nuclear 1-alfa del Hepatocito/genética , MicroARNs/uso terapéutico , Proteínas Proto-Oncogénicas c-myc/genética , ARN de Planta/uso terapéutico , Animales , Carcinogénesis/genética , Carcinogénesis/patología , Línea Celular Tumoral , Neoplasias del Colon/genética , Neoplasias del Colon/patología , Regulación hacia Abajo , Terapia Genética , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , ARN de Planta/genéticaRESUMEN
Objective: Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) are essential for vascular remodeling. Natural compounds with diterpene chinone or phenolic acid structure from Salvia miltiorrhiza, an eminent medicinal herb widely used to treat cardiovascular diseases in China, can effectively attenuate vascular remodeling induced by vascular injury. However, it remains unknown whether Salvia miltiorrhiza-derived miRNAs can protect VSMCs from injury by environmental stimuli. Here, we explored the role and underlying mechanisms of Salvia miltiorrhiza-derived Sal-miR-1 and 3 in the regulation of VSMC migration and monocyte adhesion to VSMCs induced by thrombin. Methods: A mouse model for intimal hyperplasia was established by the ligation of carotid artery and the injured carotid arteries were in situ-transfected with Sal-miR-1 and 3 using F-127 pluronic gel. The vascular protective effects of Sal-miR-1 and 3 were assessed via analysis of intimal hyperplasia with pathological morphology. VSMC migration and adhesion were analyzed by the wound healing, transwell membrane assays, and time-lapse imaging experiment. Using loss- and gain-of-function approaches, Sal-miR-1 and 3 regulation of OTUD7B/KLF4/NMHC IIA axis was investigated by using luciferase assay, co-immunoprecipitation, chromatin immunoprecipitation, western blotting, etc. Results:Salvia miltiorrhiza-derived Sal-miR-1 and 3 can enter the mouse body after intragastric administration, and significantly suppress intimal hyperplasia induced by carotid artery ligation. In cultured VSMCs, these two miRNAs inhibit thrombin-induced the migration of VSMCs and monocyte adhesion to VSMCs. Mechanistically, Sal-miR-1 and 3 abrogate OTUD7B upregulation by thrombin via binding to the different sites of the OTUD7B 3'UTR. Most importantly, OTUD7B downregulation by Sal-miR-1 and 3 attenuates KLF4 protein levels via decreasing its deubiquitylation, whereas decreased KLF4 relieves its repression of transcription of NMHC IIA gene and thus increases NMHC IIA expression levels. Further, increased NMHC IIA represses VSMC migration and monocyte adhesion to VSMCs via maintaining the contractile phenotype of VSMCs. Conclusions: Our studies not only found the novel bioactive components from Salvia miltiorrhiza but also clarified the molecular mechanism underlying Sal-miR-1 and 3 inhibition of VSMC migration and monocyte adhesion to VSMCs. These results add important knowledge to the pharmacological actions and bioactive components of Salvia miltiorrhiza. Sal-miR-1 and 3-regulated OTUD7B/KLF4/NMHC IIA axis may represent a therapeutic target for vascular remodeling.
Asunto(s)
MicroARNs/farmacología , ARN de Planta/farmacología , Salvia miltiorrhiza/genética , Túnica Íntima/patología , Remodelación Vascular/efectos de los fármacos , Animales , Arterias Carótidas/citología , Arterias Carótidas/patología , Adhesión Celular/efectos de los fármacos , Línea Celular , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Regulación hacia Abajo , Endopeptidasas/metabolismo , Humanos , Hiperplasia/tratamiento farmacológico , Hiperplasia/patología , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/metabolismo , Masculino , Ratones , MicroARNs/uso terapéutico , Monocitos/efectos de los fármacos , Monocitos/fisiología , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/fisiología , Cadenas Pesadas de Miosina/metabolismo , ARN de Planta/uso terapéutico , Transducción de Señal/efectos de los fármacos , Túnica Íntima/efectos de los fármacosRESUMEN
An increasing body of literature is addressing the immuno-modulating functions of miRNAs which include paracrine signaling via exosome-mediated intercellular miRNA. In view of the recent evidence of intake and bioavailability of dietary miRNAs in humans and animals we explored the immuno-modulating capacity of plant derived miRNAs. Here we show that transfection of synthetic miRNAs or native miRNA-enriched fractions obtained from a wide range of plant species and organs modifies dendritic cells ability to respond to inflammatory agents by limiting T cell proliferation and consequently dampening inflammation. This immuno-modulatory effect appears associated with binding of plant miRNA on TLR3 with ensuing impairment of TRIF signaling. Similarly, in vivo, plant small RNAs reduce the onset of severity of Experimental Autoimmune Encephalomyelities by limiting dendritic cell migration and dampening Th1 and Th17 responses in a Treg-independent manner. Our results indicate a potential for therapeutic use of plant miRNAs in the prevention of chronic-inflammation related diseases.
Asunto(s)
Fragaria/genética , Factores Inmunológicos/uso terapéutico , MicroARNs/uso terapéutico , ARN de Planta/uso terapéutico , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Animales , Secuencia de Bases , Proliferación Celular/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Encefalomielitis Autoinmune Experimental/patología , Femenino , Humanos , Factores Inmunológicos/farmacología , Inflamación/patología , Metilación , Ratones Endogámicos C57BL , MicroARNs/genética , Transducción de Señal/efectos de los fármacos , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Receptor Toll-Like 3/metabolismoRESUMEN
Influenza A viruses (IAVs), particularly H1N1, H5N1 and H7N9, pose a substantial threat to public health worldwide. Here, we report that MIR2911, a honeysuckle (HS)-encoded atypical microRNA, directly targets IAVs with a broad spectrum. MIR2911 is highly stable in HS decoction, and continuous drinking or gavage feeding of HS decoction leads to a significant elevation of the MIR2911 level in mouse peripheral blood and lung. Bioinformatics prediction and a luciferase reporter assay showed that MIR2911 could target various IAVs, including H1N1, H5N1 and H7N9. Synthetic MIR2911 significantly inhibited H1N1-encoded PB2 and NS1 protein expression, but did not affect mutants in which the MIR2911-binding nucleotide sequences were altered. Synthetic MIR2911, extracted RNA from HS decoction and HS decoction all significantly inhibited H1N1 viral replication and rescued viral infection-induced mouse weight loss, but did not affect infection with a mutant virus in which the MIR2911-binding nucleotide sequences of PB2 and NS1 were altered. Importantly, the inhibitory effect of HS decoction on viral replication was abolished by an anti-MIR2911 antagomir, indicating that the physiological concentration of MIR2911 in HS decoction could directly and sufficiently suppress H1N1 viral replication. MIR2911 also inhibited H5N1 and H7N9 viral replication in vitro and in vivo. Strikingly, administration of MIR2911 or HS decoction dramatically reduced mouse mortality caused by H5N1 infection. Our results demonstrate that MIR2911 is the first active component identified in Traditional Chinese Medicine to directly target various IAVs and may represent a novel type of natural product that effectively suppresses viral infection.