A Network Pharmacology-Based Study on Antidepressant Effect of Salicornia europaea L. Extract with Experimental Support in Chronic Unpredictable Mild Stress Model Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the pharmacodynamic material basis, mechanism of actions and targeted diseases of Salicornia europaea L. (SE) based on the network pharmacology method, and to verify the antidepressant-like effect of the SE extract by pharmacological experiments.@*METHODS@#Retrieval tools including Chinese medicine (CM), PubMed, PharmMapper, MAS 3.0 and Cytoscape were used to search the components of SE, predict its targets and related therapeutic diseases, and construct the "Component-Target-Pathway" network of SE for central nervous system (CNS) diseases. Further, protein-protein interaction (PPI) network, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) function annotation of depression-related targets were analyzed to predict the antidepressant mechanism of SE. Chronic unpredictable mild stress (CUMS) model was used to construct a mouse model with depression-like symptoms. And the animals were randomly divided into 6 groups (n=10) including the normal group (nonstressed mice administered with distilled water), the CUMS group (CUMS mice administered with distilled water), the venlafaxine group (CUMS mice administered with venlafaxine 9.38 mg/kg), SE high-, medium-, and low-dose groups (CUMS mice administered with SE 1.8, 1.35 and 0.9 g/kg, respectively). Then some relevant indicators were determined for experimental verification by the forced swim test (FST), the tail suspension test (TST) and open-field test (OFT). Dopamine (DA) concentration in hippocampus and cerebral cortex, IL-2 and corticosterone (CORT) levels in blood, and nuclear factor E2 related factor 2 (Nrf2), kelch-like epichlorohydrin related protein 1 (Keap1), NAD(P) H dehydrogenase [quinone] 1 (NQO1) and heme oxygenase-1 (HO-1) levels in mice were measured by enzyme linked immunosorbent assay (ELISA) and Western blot respectively to explore the possible mechanisms.@*RESULTS@#The "target-disease" network diagram predicted by network pharmacology, showed that the potential target of SE involves a variety of CNS diseases, among which depression accounts for the majority. The experimental results showed that SE (1.8, 1.35 g/kg) significantly decreased the immobility period, compared with the CUMS group in FST and TST in mice after 3-week treatment, while SE exhibited no significant effect on exploratory behavior in OFT in mice. Compared with CUMS group, the SE group (0.9 g/kg) showed significant differences (P<0.05) in DA levels in the hippocampus and cerebral cortex. In addition, compared with CUMS control group, SE (1.8 g/kg) group showed a significant effect on decreasing the activities of CORT (P<0.05), and serum IL-2 level with no statistical significance. Finally, Western blot results showed that compared with the model group, Nrf2, Keap1, NQO1 and HO-1 protein expressions in SE group (1.8 g/kg) were up-regulated (all P<0.01).@*CONCLUSION@#The SE extract may have an antidepressant effect, which appeared to regulate Nrf2-ARE pathway and increased levels of DA and CORT in the hippocampus and cortex.

The mechanism of active ingredients in Salicornia europaea in treatment of diabetes based on pharmacological network / 中成药

AIM To develop a pharmacological network screening method in predicting the potential target,active ingredients and pathway of Salicornia europaea L.for the treatment of diabetes,and to uncover its underlying multi-component,multi-target,multi-pathway mechanism.METHODS Information about fifteen kinds of bioactive chemical constituents of Salicornia europaea L.acquired from a large amount of literature were used to predict the targets according to PharmMapper Server,and such a prediction was also subjected to the screening of the antidiabetes drug targets approved by FDA in the DrugBank database.The relevant information of potential target and pathway was obtained by MAS 3.0 biomolecule function software.Cytoscape software was used to construct the Salicornia europaea L.ingredients-targets-pathways network.RESULTS Fifteen major active ingredients of Salicornia europaea L.affecting in a total of 86 pathways (VEGF signaling pathway,Fc epsilon RI signaling pathway,T cell receptor signaling pathway,etc),including the 30 particular diabetes-related pathways of MAP2K1,MAPK,GSK3B,AKT,etc.,fully demonstrated the multi-component,multi-target,multi-pathway mechanism of Salicornia europaea L.in the treatment of diabetes and its complications,through regulating immune,lipid metabolism,inflammation,apoptosis and other processes.CONCLUSION Given the new understanding in analyzing the scientific connotation of anti-diabetes effect,and the complex system of Salicornia europaea L.,this paper highlights the direction for the next step in the validation experiment of its target and mechanism.

A choline monooxygenase gene promoter from Salicornia europaea increases expression of the -glucuronidase gene under abiotic stresses in tobacco (Nicotiana tabacum L.).

A 1312 bp 5' flanking region of Salicornia europaea choline monooxygenase (SeCMO) gene was isolated using the anchored PCR. To investigate the mechanism of regulation for this stress-induced gene, the SeCMO promoter--glucuronidase (GUS) chimeric gene constructs containing five deletions F1, F2, F3, F4 and F5 were introduced into tobacco (Nicotiana tabacum L.) by Agrobacterium-mediated transformation. The functional properties of each promoter fragment were examined by assaying GUS activity in the leaves of transgenic tobacco treated with abiotic stresses (NaCl, PEG6000 and low temperature). The GUS activity in transgenic tobacco with F2 (-1056 to +8) construct showed highest increase under all the three abiotic stresses. Thus, the study provided a potential promoter induced by the salt, dehydration and cold for the plant genetic manipulation.

Isolation of choline monooxygenase (CMO) gene from Salicornia europaea and enhanced salt tolerance of transgenic tobacco with CMO genes.

Glycinebetaine (GB) is an osmoprotectant accumulated by certain plants in response to high salinity, drought, and cold stress. Plants synthesize GB via the pathway choline → betaine aldehyde → glycinebetaine, and the first step is catalyzed by choline monooxygenase (CMO). In the present study, by using RT-PCR and RLM-RACE, a full-length CMO cDNA (1844 bp) was cloned from a halophyte Salicornia europaea, which showed high homology to other known sequences. In order to identify its function, the ORF of CMO cDNA was inserted into binary vector PBI121 to construct the chimeric plant expression vector PBI121-CMO. Using Agrobacterium (LBA4404) mediation, the recombinant plasmid was transferred into tobacco (Nicotiana tabacum). The PCR, Southern blot and RT-PCR analysis indicated the CMO gene was integrated into the tobacco genome, as well as expressed on the level of transcription. The transgenic tobacco plants were able to survive on MS medium containing 300 mmol/L NaCl and more vigorous than those of wild type with the same concentration salt treatment. In salt-stress conditions, transgenic plants had distinctly higher chlorophyll content and betaine accumulation than that of the control, while relative electrical conductivity of transgenic plants was generally lower. The results suggested the CMO gene transformation could effectively contribute to improving tobacco salt-resistance.