摘要
目的研究环木菠萝甾醇阿魏酸(cycloartenyl ferulate, CF)对脂多糖(lipopolysaccharide, LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)凋亡的影响及机制。方法体外培养HUVECs,分为正常组、CF组、LPS组、LPS+CF不同剂量组。各组细胞经过相应处理后,通过采用CCK-8法检测细胞活力,采用TUNEL法观察细胞凋亡情况。同时采用Western blot检测细胞Bax、Bcl-2和Caspase-3蛋白的表达情况以及CF对Nrf2/HO-1信号通路蛋白表达水平的影响。使用SPSS 23.0统计软件,多组间的比较采用单因素方差分析,各组均数间两两比较采用LSD-t检验。结果与LPS组相比,不同剂量CF组HUVECs细胞存活率明显增加,且与CF的剂量呈正向依赖性,其中320 μmol/L CF组增加显著[(69.85±1.2)% vs ( 100.2±1.824)%, P<0.01]。TUNEL染色法显示,与LPS组相比,320 μmol/L CF组凋亡阳性细胞数明显减少[(27.33±3.40) vs (11.67±2.04),P<0.01]。与LPS组相比,不同剂量CF组中Bcl-2的蛋白表达均显著升高(P<0.01),Caspase-3和Bax蛋白表达均明显下调(均P<0.01), Nrf2蛋白表达水平升高明显(P<0.01),HO-1蛋白水平升高(P<0.01)。结论CF可减轻LPS诱导的HUVECs凋亡,其作用机制可能是通过增加Bcl-2表达,减少Caspase-3和Bax蛋白表达,并激活Nrf2/HO-1信号通路有关。
Objective To investigate the effect of cycloartenyl ferulate (CF) on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVEC), and to explore its relative mechanism. Methods Human umbilical vein endothelial cells were cultured in vitro, and the experiment was divided into the normal group, CF group, LPS group, and LPS+CF groups at different concentrations. After the corresponding treatment of cells in each group, the cell viability and apoptosis of each group were tested by the cell counter Kit-8 (CCK-8) assay and TdT-mediated dUTP nick end labeling (TUNEL) assay. The protein expression of Bax, Bcl-2, caspase-3 and the effect of CF on the protein expression of Nrf2/HO-1 pathway were determined by Western blot. One-way ANOVA were performed in multigroup mean comparison, LSD-t test was used to compare the mean values of two samples, and P<0.05 was considered statistically significant. Results Compared with the LPS group, the survival rate of HUVECs cells in the CF group was significantly increased with different doses, and the survival rate increased significantly in the 320 μmol/L CF group [(69.85±1.2)% vs ( 100.2±1.824)%, P< 0.01]. TUNEL staining showed that compared with the LPS group, the number of apoptotic positive cells in the 320 μmol/L CF group was significantly reduced [(27.33 ± 3.40) vs (11.67 ± 2.04), P<0.01]. Compared with the LPS group, Bcl-2 protein expression level was significantly increased in the CF group at different doses (P<0.01), caspase-3 and Bax protein expression were significantly decreased (P<0.01). Nrf2 protein expression level increased significantly (P<0.01), and HO-1 protein level increased significantly (P<0.01). Conclusion CF can alleviate LPS-induced apoptosis of HUVEC, which may be related to the increase of bcl-2 expression, the decrease of caspase-3 and Bax protein expression and the activation of Nrf2/ HO-1 signaling pathway.
作者
倪菁晶
徐荣校
杨玺渝
陈辉
陈隆望
连洁
赵光举
卢中秋
Ni Jingjing;Xu Rongxiao;Yang Xiyu;Chen Hui;Chen Longwang;Lian Jie;Zhao Guangju;Lu Zhongqiu(Emergency Department, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China)
出处
《中华急诊医学杂志》
CAS
CSCD
北大核心
2019年第9期1106-1111,共6页
Chinese Journal of Emergency Medicine
基金
浙江省医药卫生重大科技计划(WKJ-ZJ-1724)
浙江省中医药管理局项目(2015ZZ015).
