论文部分内容阅读
目的:探讨在脂多糖(LPS)诱导的条件下中国蜂胶对血管内皮细胞(VECs)磷脂酰胆碱特异性磷脂酶C(PC-PLC)活性和TLR4表达的影响。方法:将100μg/L LPS加入到含0.5%血清的培养液中培养VECs,经12.5 mg/L的中国蜂胶分别处理12 h和24 h后,SRB法测定细胞存活率;化学法测定NO含量;以L-α-卵磷脂为底物测定PC-PLC的活性;Western blotting检测TLR4、核因子κB p65(NF-κB p65)和p53的表达;细胞内活性氧(ROS)和线粒体膜电位分别通过荧光探针DCHF和JC-1检测。结果:中国蜂胶处理LPS诱导的血管内皮细胞24h并不影响细胞存活率,但降低NO的含量和ROS的水平;处理12 h后降低PC-PLC活性和NF-κB p65表达;处理12 h和24 h后降低TLR4和p53的表达。此外,中国蜂胶不影响细胞内线粒体膜电位的水平。结论:中国蜂胶通过降低PC-PLC活性和TLR4表达,抑制其下游信号分子NF-κB p65、p53的表达和ROS的水平,以及抑制NO的释放,从而发挥抗炎功效。
Objective: To investigate the effect of propolis on the activity of phosphatidylcholine-specific phospholipase C (PC-PLC) and the expression of TLR4 in vascular endothelial cells (VECs) induced by lipopolysaccharide (LPS). Methods: VECs were cultured in culture medium containing 0.5% serum with 100μg / L LPS. After treated with 12.5mg / L Chinese propolis for 12 and 24 hours respectively, the cell survival rate was determined by SRB method. The content of NO was determined by chemical method. The activity of PC-PLC was determined by L-α-lecithin. The expression of TLR4, NF-κB p65 and p53 were detected by Western blotting. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential Fluorescent probes DCHF and JC-1 detection. Results: LPS-treated vascular endothelial cells treated with propolis for 24 h in China did not affect cell viability, but decreased NO and ROS levels. PC-PLC activity and expression of NF-κB p65 decreased after 12 h of treatment; h to reduce the expression of TLR4 and p53. In addition, Chinese propolis does not affect the level of mitochondrial membrane potential in cells. CONCLUSION: Chinese propolis exerts anti-inflammatory effects by decreasing PC-PLC activity and TLR4 expression, inhibiting the expression of downstream signaling molecules NF-κB p65, p53 and ROS, and inhibiting the release of NO.