Sal B attenuates rat hepatocytes injury induced by hypoxia/reoxygenation via modulation of SIRT1/NF-κB/p53 pathway / 中国药理学通报

Aim To investigate the effect of salvianolic acid B(Sal B) on the attenuation of rat hepatocyte in-jury induced by hypoxia/reoxygenation(H/R) and its possible molecular mechanism. Methods Rat hepato-cytes BRL-3A were cultured in vitro. H/R injury mod-el was established and then BRL-3A cells were pretrea-ted with Sal B. The viability of cells was measured by CCK-8 assay;the expression of ALT and AST was de-tected by microplate assay; the levels of TNF-α and IL-1β were determined by ELISA; the apoptosis was detected by flow cytometry;the protein and mRNA lev-els of SIRT1, NF-κB p65, p53, Bax and Bcl-2 were measured by Western blot and qPCR. Results H/R intervention decreased the viability and increased the apoptosis of cells;the production of ALT, AST, TNF-α and IL-1β was elevated;the protein and mRNA lev-els of SIRT1, Bcl-2 were reduced, but the levels of NF-κB p65, p53 and Bax increased. After pretreated with Sal B, the viability of cells increased while the apoptosis decreased; the expression of ALT, AST, TNF-α and IL-1β was inhibited;moreover,the protein and mRNA levels of SIRT1,Bcl-2 were enhanced,and the levels of NF-κB p65, p53 and Bax decreased sig-nificantly. Conclusion Sal B may attenuate rat hepa-tocyte injury induced by H/R via the SIRT1/NF-κB/p53 pathway.

Effect of procyanidine on the function and expression of miR-221 in human umbilical vein endothelial cells / 解放军医学杂志

Objective To study the effect ofprocyanidine (PC) on the proliferation and migration of human umbilical vein endothelial cells (HUVECs),determine the expression changes of miR-221,and to investigate the mechanism involved.Methods HUVECs were cultured in vitro and treated with PC (5,25,50,75,100μg/ml) for 24 hours,and a PC concentration of 50μg/ml was screened by CCK-8 assay for the follow up experiment,then the cell proliferation activity was detected by the wound healing assay.The expression of miR-221 in HUVECs was detected by real-time quantitative PCR;MiR-221 target genes were predicted in miRWalk database,and the target genes were analyzed by GO and KEGG pathway.Results Compared with the control group,the HUVECs treated with PC for 24h,their proliferation activity in PC 5μg/ml group did not change obviously (P＞0.05),and in PC 25,50,75 and 100μg/ml groups decreased in a concentration dependent manner (P＜0.01).Compared with the control group,the migration ability of PC 50μg/ml group decreased markedly (P＜0.01).Compared with the control group,the expression of miR-221 increased after treatment with PC 50μg/ml (P＜0.01).Go analysis indicated that the target genes of miR-221 were mainly related to cell proliferation,migration,gene translation and so on.The target genes related to cell proliferation and migration were ADAM17,KIT,PDGFD and so on.KEGG pathway analysis showed that the target genes of miR-221 enriched 5 signal pathways,such as FoxO,PI3K-Akt and so on.Conclusions Low concentration of PC has no effect on the proliferation activity of HUVECs.A certain concentration of PC can inhibit the proliferation and migration of HUVECs,of which the mechanism may be involved with the up-regulation of miR-221 and FoxO,PI3K-Akt and other signaling pathways.

Intervention effects of Jiaotai pills on PCPA-induced insomnia in rats / 中国中药杂志

To elucidate the intervention effects of Jiaotai pills(JTP) on p-chlorophenylalanine (PCPA)-induced insomnia in rats and its underlying mechanism, the insomnia model was established by single intraperitoneal injection with PCPA in rats. The locomotor activity of rats was observed, and the levels of nerve growth factor(NGF) in hypothalamus, hippocampus, prefrontal cortex and serum of rats were determined by using ELISA. Moreover, a proton nuclear magnetic resonance(¹H-NMR)-based metabonomic approach was developed to profile insomnia-related metabolites in rat serum and hippocampus and analyze the intervention effects of JTP on changes in underlying biomarkers related to locomotor activity, NGF and insomnia. According to the results, JTP could significantly suppress the locomotor activity of insomnia rats, and increase the NGF levels in hypothalamus, hippocampus, prefrontal cortex and serum of rats with insomnia. The disturbed metabolic state associated with PCPA-induced insomnia in rat serum and hippocampus could be intervened by JTP. Meanwhile, six and five potential biomarkers related to insomnia in rat serum and hippocampus were reversed by administration of JTP. In conclusion, the current study demonstrated that JTP had protective effects against PCPA-induced insomnia in rats, which was probably correlated with regulation of NGF level and metabolism of amino acids, lipids and choline.