摘要
目的探讨二苯乙烯苷（TSG）对血清诱导的血管平滑肌细胞（VSMCs）增殖的影响,并研究其相关机制。方法组织贴块法原代培养大鼠主动脉VSMCs,采用5-溴脱氧尿嘧啶核苷（Brdu）参入法检测VSMCs增殖情况,乳酸脱氢酶（LDH）法检测药物对细胞毒性作用,流式细胞仪测定细胞周期,Western blot法检测增殖细胞核抗原（PCNA）的蛋白表达,荧光法测定胞内活性氧（ROS）水平,酶生化法测定细胞内超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-px）活性。结果 0.1-100μmol.L^-1实验浓度的TSG无细胞毒性作用;10μmol.L^-1和100μmol.L^-1TSG可明显抑制血清诱导的VSMCs增殖,抑制细胞由G0/G1期向S期的转变,抑制血清诱导的VSMCs核内PCNA的蛋白表达,降低细胞内ROS含量;100μmol.L^-1TSG能明显提高SOD、GSH-px活性。结论一定浓度的TSG能抑制血清诱导的VSMCs增殖,其机制可能与TSG影响细胞增殖周期、抑制PCNA的表达、提高VSMCs抗氧化能力有关。
Aim To study the effect of 2,3,4′,5-tetrahydroxystilbene-2-O-β-D glucoside（TSG） on vascular smooth muscle cells（VSMCs） proliferation induced by fetal bovine serum and explore the underlying mechanisms.Methods VSMCs were cultured from rat aorta using explant technique.VSMCs proliferation was analyzed by 5-bromo-2-deoxyuridine（Brdu） incorporation assay.Cell cycle phase distributions were determined by flow cytometry.The expression of proliferating cell nuclear antigen（PCNA） was evaluated by Western blot analysis.The level of intracellular reactive oxygen species（ROS） was estimated through fluorescence assay.The intracellular activities of superoxide dismutase（SOD）,catalase（CAT）,and glutathi one peroxidase（GSH-px） were measured using biochemical method.Results TSG in the experimental concentration 0.1～100 μmol·L-1 exhibited no apparent cytotoxocity.10 μmol·L-1 and 100 μmol·L-1 TSG significantly inhibited the proliferation of VSMCs induced by serum,cell cycle transition from G0/G1phase to S phase,PCNA expression in nucleus of VSMCs and level of intracellular ROS.100 μmol·L-1 TSG markedly increased the activities of SOD and GSH-px.Conclusions Certain concentration of TSG can prevent VSMCs proliferation effect mediated by serum;the mechanism about TSG effect may be associated with arresting G0/G1 to S progression,decreasing PCNA expression and improving the antioxidation of VSMCs.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第7期934-939,共6页
Chinese Pharmacological Bulletin
基金
江苏省自然科学基金资助项目(NoBK2009162)
江苏省高校自然科学研究项目(No09KJB310010)
南通大学引进人才启动费项目(No09R15)
关键词
二苯乙烯苷
血管平滑肌细胞
增殖
细胞周期
增殖细胞核抗原
氧化应激
2
3
4′
5-tetrahydroxystilbene-2-O-β-D glucoside
vascular smooth muscle cells
proliferation
cell cycle
proliferating cell nuclear antigen
oxidative stress
