摘要
目的明确灯盏花素(scutellarin,Scu)对缺血/再灌注损伤(I/R)微血管内皮细胞(MEC)氧化应激、自噬及凋亡的作用并研究可能的机制。方法MEC制作I/R模型,MTT法测细胞活力;生化法测定SOD、MDA及LDH;RT-PCR测定ICAM-1、VCAM-1、caspase-3及LC3的mRNA表达;Western blot测定LC3、CREB及TLR4表达。结果50μmol/L灯盏花素使I/R损伤MEC的细胞活力增加(P<0.01),SOD表达增加(P<0.01),MDA表达减少(P<0.05),LDH减少(P<0.05),ICAM-1(P<0.01)及VCAM-1(P<0.05)的mRNA表达减少,LC3 mRNA表达增加(P<0.05),caspase-3 mRNA表达减少(P<0.01),LC3和CREB蛋白表达增加(P<0.05),TLR4蛋白表达减少(P<0.05)。抑制CREB表达使灯盏花素治疗的I/R细胞LC3表达降低(P<0.05)。激动TLR4表达使灯盏花素治疗的I/R细胞caspase-3 mRNA表达增加(P<0.05)。结论灯盏花素可能是通过促进CREB表达促进自噬,通过抑制TLR4表达对抗氧化损伤、抑制凋亡、保护内皮细胞I/R损伤。
Aim To investigate the effects of Scutellarin(Scu)on microvascular endothelial cells(MEC)with ischemia/reperfusion injury(I/R)and its mechanisms involved.Methods Microvascular endothelial cells were used to build model of I/R,and the protective effects of Scu were observed.Cell viability was determined by MTT assay;SOD,MDA and LDH levels were determined by biochemical method;ICAM-1,VCAM-1,caspase-3 and LC3 mRNA expressions were detected by RT-PCR;LC3,CREB and TLR4 expressions were detected by Western blot.Results MTT assay showed that the cell viability was rescued by 50μmol·L-1 Scu(P<0.01).Compared with I/R group,SOD expression increased(P<0.01),while MDA and LDH expression decreased in Scu treatment group(P<0.05),decreased mRNA expression of ICAM-1(P<0.01),VCAM-1(P<0.05)and caspase-3 mRNA(P<0.01),meanwhile increased mRNA expression of LC3(P<0.05)was observed in Scu treatment group;the protein expressions of CREB and LC3 increased(P<0.05),the protein expression of TLR4 decreased(P<0.05)in Scu treatment group.Using CREB inhibitor(KG-501)in Scu+I/R group,LC3 expression decreased(P<0.05).Agonist of TLR4(LPS)increased the caspase-3 mRNA expression in Scu+I/R group(P<0.05).Conclusions Scutellarin might be against oxidative damage and anti-apoptosis by inhibiting TLR4 expression,and promote autophagy by promoting CREB expression in endothelial cells.
作者
王娓娓
叶宏
孙林
鲍天昊
WANG Wei-wei;YE Hong;SUN Lin;BAO Tian-hao(Cardiology Department, the Second Affiliated Hospital of Kunming Medical University,Kunming 650101,China;Pathology Department, Kunming Medical University,Kunming 650101,China;Geriatrics Department, Yunan Psychiatric Hospital,Kunming 650101,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第7期946-951,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金地区项目(No 81860236)
云南省科技厅-昆明医科大学联合专项基金[No 2018FE001(-148),2018FE001(-223)]。
关键词
灯盏花素
缺血/再灌注损伤
自噬
凋亡
内皮细胞
氧化损伤
Scutellarin
ischemia-reperfusion injury
autophagy
apoptosis
endothelial cells
oxidative injury
