Effect of Xinnao Shutong capsule and its active ingredients on damage of cerebral microvascular endothelial cell barrier integrity caused by oxygen glucose deprivation/reoxygenation and its mechanism / 中国药理学通报

Aim To investigate the effect of XNST and its monomeric components on the barrier structure and tight junction protein expression of brain microvascular endothelial cells damaged by oxygen glucose deprivation/reoxygenation (OGD/R) and the possible mechanism. Methods The mouse brain microvascular endothelial cell line bEnd. 3 was inoculated in the upper layer of the Transwell chamber to establish an OGD/R damage model, and the effect of the drug on the integrity of the endothelial cell barrier was investigated by the transmembrane resistance value and fluorescein-so-dium transmittance. Claudin-5 immunofluorescence staining was used to observe the changes of tight junction structure between endothelial cells. RT-PCR and Western blot were employed to detect mRNA and protein expression levels of tightly linked proteins Claudin-5 , Occludin, ZO-1. Western blot was applied to detect the expression levels of MAPKs (JNK, p38, ERK) , I kappa B a, I kappa B kinase phosphorylated protein expression, and Western blot and immunofluorescence were utilized to detect NF-K.B/p65 nucleation expression. Results XNST and its three monomers could significantly increase endothelial cell resistance and de- crease fluorescein-sodium transmittance. Claudin-5 fluorescence staining showed that the tight junction between cells in the model group was significantly damaged , while XNST and its monomer components could significantly improve its tight structure. RT-PCR and Western blot results showed that it could significantly upregulate the expression of mRNA and protein of Claudin-5, Occludin and ZO-1, and further study on the mechanism showed that XNST and its monomer components could significantly inhibit the phosphoryla-tion of JNK, p38 and ERK, inhibit the phosphorylation of I kappa B a and I kappa B kinases, and significantly inhibit the nuclear translocation of NF-KB/p65. Conclusion Both XNST and its monomeric components can exert cerebroprotective effects by increasing the tight junction structure between cells to promote barrier integrity, and the mechanism may be related to inhibition of NF-kB and MAPKs signaling pathway activation.

3,6-dihydroxyflavone decreases anti-oxidative enzyme activities in human leukemia HL-60 cells via MAPKs signaling pathway / 第三军医大学学报

Objective To investigate the intracellular mechanisms by which 3,6-dihydroxyflavone inducing apoptosis of HL-60 cells.Methods After the HL-60 cells were treated with 3,6-dihydroxyflavone at the concentration of 5.10,20 or 40 mol/L,the cells then were inoculated under the selected optimal concentration.The survival rate of HL-60 cells was analyzed using a haemocytometer with standard trypan blue dye exclusion and a CASY cell counter and analyzer.The contents of malondialdehyde(MDA)and the activities of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)in the cells were measured by chemochromatometry.The expression of phosphorylation of ERK,JNK and p38MAPK were examined by Western blotting.Results The survival rate of HL-60 cells was significantly reduced in a dose-and time-dependent manner after 3,6-dihydroxyflavone treatment.When the dose was over 10 mol/L or the time was longer than 12 h under the concentration of 20 mol/L,significantly decreased survival rate was observed(P