RESUMO
Strain 39 is an endophytic fungus which was isolated from Dioscorea nipponica Makino (DNM). After Strain 39 co-cultured with ethanol extract of DNM rhizomes for several days, the content of saponins in this culture mixture would be obviously increased. To analyze the mechanism of this microbial transformation, we used the differential display reverse transcription polymerase chain reaction (DDRT-PCR) method to compare the transcriptomes between Strain 39 cultured in normal PD medium and in PD medium which added ethanol extract of DNM rhizomes. We amplified 29 DDRT-PCR bands using 12 primer combinations of three anchored primers and five random primers, and six bands were re-amplified. Analysis of real-time PCR and sequence alignment showed that three clones were up-regulated in sample group: squalene epoxidase, squalene synthase, and catalase, one clone was expressed only in sample group. The possible roles and origins of the above genes were discussed, and the molecular mechanism of Strain 39 biotransformation was speculated. This study is the first report of the molecular biotransformation mechanism of saponins production by endophytic fungus of DNM.
Assuntos
Dioscorea/microbiologia , Endófitos/genética , Proteínas Fúngicas/genética , Fungos/genética , Dioscorea/química , Endófitos/classificação , Endófitos/isolamento & purificação , Endófitos/metabolismo , Proteínas Fúngicas/metabolismo , Fungos/classificação , Fungos/isolamento & purificação , Fungos/metabolismo , Regulação Fúngica da Expressão Gênica , Extratos Vegetais/metabolismo , Saponinas/metabolismoRESUMO
Panax ginseng is one of the oldest and most generally prescribed herbs in Eastern traditional medicine to treat diseases. Several studies had documented that ginseng leaves have anti-oxidative, anti-inflammatory, and anticancer properties similar to those of ginseng root. The aim of this research was to forecast of the molecular mechanism of ginseng leaves on lung cancer by molecular docking and network pharmacology so as to decipher ginseng leaves' entire mechanism. The compounds associated with ginseng leaves were searched by TCMSP. TCMSP and Swiss Target Prediction databases were used to sort out the potential targets of the main chemical components. Targets were collected from OMIM, PharmGKB, TTD, DrugBank and GeneCards which related to immunity and lung cancer. Ginseng leaves exert its lung cancer suppressive function by regulating the several signaling proteins, such as JUN, STAT3, AKT1, TNF, MAPK1, TP53. GO and KEGG analyses indicated that the immunoreaction against lung cancer by ginseng leaves might be related to response to lipopolysaccharide, response to oxidative stress, PI3K-Akt, MAPK and TNF pathway. Molecular docking analysis demonstrated that hydrogen bonding was interaction's core forms. The results of CCK8 test and qRT-PCR showed that ginseng leaves inhibit cell proliferation and regulates AKT1 and P53 expression in A549. The present study clarifies the mechanism of Ginseng leaves against lung cancer and provides evidence to support its clinical use.
Assuntos
Antineoplásicos/farmacologia , Fatores Imunológicos/farmacologia , Neoplasias Pulmonares/metabolismo , Panax/química , Extratos Vegetais/farmacologia , Células A549 , Proliferação de Células/efeitos dos fármacos , Humanos , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Folhas de Planta/química , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
OBJECTIVE: To study the pharmacological effects of volatile oil of Valeriana amurensis on central nervous system. METHODS: The pharmacological effects of volatile oil of Valeriana amurensis on the autonomic activities of mice, the sleeping synergistic action of mice with pentoharbital sodium at subthreshold and hypnotic dosages, the sleep phases of rats, the writhing response of mice caused by acetic acid and the convulsion of mice induced by thio-semicarbazide were investigated. RESULTS: The autonomic activities of mice were significantly inhibited by the volatile oil of Valeriana amurensis. The rate of falling sleep and the extension of sleeping time of mice were significantly increased by the synergic action of pentobarbital sodium with the volatile oil. The sleep phases of SWS2 and REMS of rats were obviously extended by the volatile oil of Valeriana amurensis. In addition, the frequency of writhing response of mice caused by acetic acid was reduced, and the convulsion of mice induced by thio-semicarbazide was antagonized with the administration of the volatile oil. CONCLUSION: The volatile oil of Valeriana amurensis has significantly sedative analgesic and anti-hyperspasmia effects.