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OBJECTIVE@#To study the in vitro and in vivo antitumor effects of the polysaccharide of Alocasia cucullata (PAC) and the underlying mechanism.@*METHODS@#B16F10 and 4T1 cells were cultured with PAC of 40 µg/mL, and PAC was withdrawn after 40 days of administration. The cell viability was detected by cell counting kit-8. The expression of Bcl-2 and Caspase-3 proteins were detected by Western blot and the expressions of ERK1/2 mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). A mouse melanoma model was established to study the effect of PAC during long-time administration. Mice were divided into 3 treatment groups: control group treated with saline water, positive control group (LNT group) treated with lentinan at 100 mg/(kg·d), and PAC group treated with PAC at 120 mg/(kg·d). The pathological changes of tumor tissues were observed by hematoxylin-eosin staining. The apoptosis of tumor tissues was detected by TUNEL staining. Bcl-2 and Caspase-3 protein expressions were detected by immunohistochemistry, and the expressions of ERK1/2, JNK1 and p38 mRNA were detected by qRT-PCR.@*RESULTS@#In vitro, no strong inhibitory effects of PAC were found in various tumor cells after 48 or 72 h of administration. Interestingly however, after 40 days of cultivation under PAC, an inhibitory effect on B16F10 cells was found. Correspondingly, the long-time administration of PAC led to downregulation of Bcl-2 protein (P<0.05), up-regulation of Caspase-3 protein (P<0.05) and ERK1 mRNA (P<0.05) in B16F10 cells. The above results were verified by in vivo experiments. In addition, viability of B16F10 cells under long-time administration culture in vitro decreased after drug withdrawal, and similar results were also observed in 4T1 cells.@*CONCLUSIONS@#Long-time administration of PAC can significantly inhibit viability and promote apoptosis of tumor cells, and had obvious antitumor effect in tumor-bearing mice.
Subject(s)
Mice , Animals , Alocasia/metabolism , MAP Kinase Signaling System , Caspase 3/metabolism , Apoptosis , RNA, Messenger/metabolismABSTRACT
Aim To study the effect of three kinds of active ingredients of traditional Chinese medicine (sinomenine, rhynchophylline and isorhynchophylline) on dre-miR-723-5p expression in morphine-induced zebrafish brain. Methods Morphine was injected intraperitoneally to zebrafish, conditional position preference (CPP) was trained and then the behavioral of animals were observed; the miRNA expression profiles of morphine-additive zebrafish were determined by small RNA sequencing; qRT-PCR was used to verify the expression of dre-miR-723-5p, three target gene databases (miRanda, miRDB, andRNAhybrid) were used to predict the target genes of dre-miR-723-5p; Kobas 3.0 was used to perform Gene Ontology (GO) and KEGG pathway analysis of these target genes. Results Morphine-induced CPP model was established successfully. Compared with control group, the resident time and movement map in drug-pair box of zebrafish in model group significantly increased. After drug administration, the resident time and movement map in drug-pair box of zebrafish decreased. The verification results of qRT-PCR were consistent with the results of small RNA sequencing. Ninety-nine putative target genes of dremiR-723-5p that were common to all three target gene databases, which were mainly enriched in biological process, cell composition and molecular function, involved in the positive regulation of MAPK signaling pathway, lysosome, cytokine-cytokine receptor interaction, and apoptosis. Conclusion Morphine can increase the expression of dre-miR-723-5p in the zebrafish brain, which can be reversed by sinomenine, isorhynchophylline, and rhynchophylline treatment, and dre-miR-723-5p may participate in the mechanism underlying morphine-induced damage of brain.
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Aim To explore the effect of rhynchophyl-line on the expression of tyrosine hydroxylase ( TH) in hippocampus of methamphetamine-induced condition place preference ( CPP) mice. Methods Metham- phetamine was injected intraperitoneally to mice, and the expression of TH was observed by immunohisto-chemistry and Western blot. Results The CPP mouse model was established successfully by methamphet-amine ( 4 mg·kg-1) . Ketamine ( 15 mg·kg-1) , rhynchophylline low dosage group (40 mg·kg-1) and rhynchophylline high dosage group ( 80 mg·kg-1) could remove the effect of methamphetamine on CPP mice. The result of immunohistochemistry showed that methamphetamine ( 4 mg·kg-1) could increase the number of TH positive cells in hippocampus while ket-amine (4 mg·kg-1), rhynchophylline (40, 80 mg· kg-1) group could attenuate the change. Western blot-ting indicated the expression of TH of model group in-creased significantly, whereas ketamine ( 15 mg· kg-1) , rhynchophylline ( 40, 80 mg·kg-1) group presented less expression. Conclusions The CPP in-duced by methamphetamine in mice may be inhibited to some extent by rhynchophylline, and its mechanism may be associated with the expression of TH.
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Exosomes are nanoscale vesicles produced and secre-ted into extracellular fluid by all cells. They mediate cell com-munication through carrying and transferring informational car-goes ( proteins, nucleic acids, lipids and so on ) to recipient cells. In central nervous system, exosomes can be released from all cell types including neurons, neural stem cells and neuroglia cells. These exosomes shuttle nucleic acids ( miRNAs, mRNAs and so on) and play an important role in nervous system devel-opment and function as well as diseases including Alzheimer's disease and drug addiction. Furthermore, the functional effects and targeting characteristics of exosomes-shuttle-RNAs suggest that exosomes-shuttle-RNAs can be diagnostic and therapeutic targets. In this review, we elaborate the effects, functions and mechanisms of exosomes-shuttle-RNAs in order to gain a new recognition of CNS development and diseases.