RESUMO
We examined the effects of methyl jasmonate on aphid resistance of Chrysanthemum 'Hangbai'. Leaves of chrysanthemum cuttings were sprayed by methyl jasmonic with different concentrations (0.01, 0.05, 0.1, 0.5 and 1 mmol·L-1) and then inoculated with the aphids Macrosiphoniella sanborni. The following parameters are measured, including protective enzymes activities, defense enzyme activities, osmotic substances, secondary metabolites, and key genes of the jasmonic signaling pathway. The relationship between aphid resistance and the jasmonic signaling pathway in chrysanthemum was probed. The results showed that, under the treatments of all the five concentrations of methyl jasmonic, the activities of leaf protective enzymes and defense enzymes as well as the contents of secondary metabolite were increased in Chrysanthemum, whereas the contents of malondialdehyde and soluble sugar were reduced. Moreover, exogenous methyl jasmonate treatment induced the expression of CmAOS and CmCOI1, enhanced the content of endogenous jasmonic acid and the aphid resistance of chrysanthemum.
Assuntos
Afídeos , Chrysanthemum , Acetatos/farmacologia , Animais , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Folhas de PlantaRESUMO
To study the effect of aqueous extracts of Yiqi Jiedu formula (YQ) on the proliferation of CNE2 cells in human nasopharyngeal carcinoma, and investigate its mechanism to provide a new theoretical basis for the clinical application of YQ. CNE2 cells were treated with different concentrations (0.125, 0.25, 0.5, 0.25 g·L⻹) of YQ, positive control medicine (cisplatin 4.0 mg·L⻹), inhibitor PD98059 (50 µmol·L⻹), activator isoproterenol hydrochloride (20 µmol·L⻹), activator isoproterenol hydrochloride (ISO)+YQ 0.5 g·L⻹. Then cell labeling by using real-time analyzer (RTCA) and CCK 8 method were used to detect cell proliferation activity, and the half inhibitory concentration (IC50) was calculated. The cell cycle distribution was detected by fluorescence double dye flow cytometry PI staining, and Western blot method was used to detect the expression levels of related protein and MAPK/ERK signaling pathway. The results of RTCA and CCK-8 test showed that as compared with the control group, YQ group could effectively inhibit the proliferation of CNE2 cells (P<0.01), with a dose and time dependence, and 48 h IC50 value was 0.5 g·L⻹. The results of cell cycle showed that after 48 h of water extract treatment, the cell cycle was significantly changed, the proportion of G0/G1 was reduced, the ratio of G2/M increased, and the cell cycle was in G2/M period (P<0.01). Western blot results showed that after 48 h treatment with different concentrations of aqueous extract, cell cycle-related proteins cyclinD1, cyclinD3 and CDK2 expression levels were down-regulated; MAPK/ERK signaling pathway related protein p-c-Raf, p-MEK, p-ERK1/2 expression level significantly lower as compared with the control group (P<0.05). After adding activator and inhibitor in MAPK/ERK signaling pathway on this basis, the results showed that after adding activator ISO, cell proliferation was significantly higher than that in the Control group; the cycle related proteins cyclinD1, cyclinD3, and CDK2 expression levels were increased; at the same time, key protein p-c-Raf, p-MEK, p-ERK1/2 expression levels in the signal pathways were relatively increased. While after the addition of inhibitor PD98059, the cell proliferation was significantly lower than that in the Control group, and the expression level of corresponding protein was decreased, which was significantly different from the Control group (P<0.05). So YQ could block cell cycle and inhibit the proliferation of CNE2 cells mainly by reducing the expression of MAPK/ERK signaling pathway key protein p-c-Raf, p-MEK and p-ERK1/2.