ABSTRACT
This study aimed to investigate the molecular mechanism and protective effect of total saponins of Panax japonicas (TSPJ) on HepG2 cells apoptosis induced by palmitic acid (PA).The HepG2 cells were cultured , and divided into five groups: the control group, the model group, the high-dose group (50 mg·L⁻¹), the middle-dose group (25 mg·L⁻¹) and the low-dose group (12.5 mg·L⁻¹).The cells of the five groups were cultured continuously for 24 hours. The cell viability was measured with MTT. HepG2 cells apoptosis was detected by Hoechest staining and Annexin V-FITC/PI staining. The protein expressions of BCL-2, CHOP and TLR4 were measured with western blotting and flow cytometry analysis. The mRNA expressions of TNF-α, IL-1β, BCL-2, CHOP and GAPDH were measured with RT-PCR. The results suggested that compared with the control group, the number of HepG2 cells of the model group were reduced significantly (<0.01), while the number of apoptotic HepG2 cells were increased. Compared with the model group, the number of HepG2 cells of the high-dose group and the middle-dose group were increased significantly (<0.01), whereas the number of apoptotic HepG2 cells were reduced. Compared with the control group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the model group were significantly up-regulated (<0.01), while BCL-2 protein and mRNA expressions in the model group were significantly decreased (<0.01). Compared with the model group, TNF-α, IL-1β and CHOP mRNA expressions and CHOP and TLR4 protein expressions in the high-dose group were significantly decreased (<0.01), while BCL-2 protein and mRNA expressions in the high-dose group were significantly up-regulated (<0.01).In conclusion, TSPJ can reduce inflammation and apoptosis induced by palmitic acid, with a certain protective effect on liver cells.
Subject(s)
Humans , Apoptosis , Hep G2 Cells , Palmitic Acids , Panax , Chemistry , Phytochemicals , Pharmacology , Saponins , PharmacologyABSTRACT
Objective To detect the damage of hippocampal neurons and the changes in inflammatory cytokines in rats after cerebral ischemia-reperfusion(I/R)and compare the expressions of IL-1β,IL-6 and TNFαin hippocampal DG,CA1 and CA3 subregions.Methods The focal cerebral I/R model was induced by an intraluminal filament embolism.The SD rats were randomly divided into the sham-operated group(SHAM group)and the middle cerebral artery occlusion-reperfusion group(MCAO group).HE staining was employed to detect the damage to hippocampal DG, CA1 and CA3 subregions.The expression levels of IL-1β, IL-6 and TNFα were detected by immunofluorescence assay.Results Compared with SHAM group,hippocampal DG,CA1 and CA3 subregion neurons in MCAO group were severely damaged, with occurred inflammatory cell infiltration,and a large amount of neurons apoptosis, and the expressions of IL-1β, IL-6 and TNFαin each subregion increased significantly.At the same time, in MCAO group, the expression of inflammatory cytokines in CA1 subregion was more significant than that in DG and CA 3 subregions(P<0.05).Conclusion Cerebral I/R could cause neuronal damage, inflammatory cell infiltration, and neuronal apoptosis in the DG, CA1 and CA3 subregions of the hippocampus and increase the release of inflammatory cytokines.In MCAO group, the expression of inflammatory cytokines in CA1 subregion of hippocampus is significantly higher than that in DG and CA 3 subregions, suggesting that CA1 region is more sensitive to I/R injury.
ABSTRACT
Activated protein C (APC) is known to be beneficial on ischemia reperfusion injury in myocardium. However, the protection mechanism of APC is not fully understood. The purpose of this study was to investigate the effects and possible mechanisms of APC on myocardial ischemic damage. Artificially ventilated anaesthetized Sprague-Dawley rats were subjected to a 30 min of left anterior descending coronary artery occlusion followed by 2 hr of reperfusion. Rats were randomly divided into four groups; Sham, I/R, APC preconditioning and postconditioning group. Myocardial infarct size, apoptosis index, the phosphorylation of ERK1/2, Bcl-2, Bax and cytochrome c genes and proteins were assessed. In APC-administrated rat hearts, regardless of the timing of administration, infarct size was consistently reduced compared to ischemia/reperfusion (I/R) rats. APC improved the expression of ERK1/2 and anti-apoptotic protein Bcl-2 which were significantly reduced in the I/R rats. APC reduced the expression of pro-apoptotic genes, Bax and cytochrome c. These findings suggest that APC produces cardioprotective effect by preserving the expression of proteins and genes involved in anti-apoptotic pathways, regardless of the timing of administration.