阿魏酸对LPS诱导的肺微血管内皮细胞凋亡的保护作用及可能机制  

Protective effect of ferulic acid on IPS-induced apoptosis of pulmonary microvascular endothelial cells and corresponding possible mechanism

作　　者：张勤芹[1] 全超[1] 刘亚军[1] 朱金凤[2] 金秋芬[1] 王智兰[1] Zhang Qinqin;Quan Chao;Liu Yajun;Zhu Jinfeng;Jin Qiufen;Wang Zhilan(Department of Critical Care Medicine,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China;Department of Respiratory and Critical Care Medicine,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China)

机构地区：[1]南通市中医院重症医学科,南通226001 [2]南通市中医院呼吸与危重症医学科,南通226001

出　　处：《国际呼吸杂志》2022年第19期1446-1451,共6页International Journal of Respiration

基　　金：南通市中医医疗联盟课题(TZYK202005);南通市卫生健康委员会科研课题(QA2021024)。

摘　　要：目的探讨miR-17在肺微血管内皮细胞(PMVECs)的凋亡中的表达及阿魏酸(FA)对细菌脂多糖(LPS)诱导的PMVECs凋亡的作用及机制。方法本研究为实验研究。用不同剂量(0、0.1、1、10、100 mg/L)的FA刺激PMVECs 24 h,采用MTT法检测细胞活性,选择合适的FA浓度进行后续试验。将PMVECs分为对照组、LPS组、FA低剂量组(LPS+0.1 mg/L FA)、FA中剂量组(LPS+1 mg/L FA)、FA高剂量组(LPS+10 mg/L FA),流式细胞仪分析检测各种细胞凋亡率,蛋白免疫印迹法检测Caspase-3、Bax蛋白、Bcl-2蛋白,实时荧光定量聚合酶链反应(qRT-PCR)检测miR-17确定FA对PMVECs凋亡的影响。将PMVECs分为对照组、LPS组、LPS+FA组(LPS+10 mg/L FA)、LPS+FA+inhibitor NC组、LPS+FA+miR-17 inhibitor组,检测各组细胞的凋亡率及凋亡蛋白、miR-17的表达以确定FA的作用靶点。结果100 mg/L FA不适合。LPS组较对照组细胞凋亡率、Caspase-3、Bax增加,Bcl-2蛋白、miR-17减少(P值均<0.05),FA低、中、高剂量组细胞较LPS组凋亡率、Caspase-3、Bax减少,Bcl-2蛋白、miR-17增加(P值均<0.05),且FA高剂量组凋亡率较低FA剂量组减少(P<0.05)。LPS+FA+miR-17 inhibitor组较LPS+FA+inhibitor NC组的miR-17表达低,且凋亡率、Caspase-3、Bax高,Bcl-2蛋白低(P值均<0.05)。结论LPS导致的炎性状态能抑制miR-17的表达,从而导致PMVECs凋亡,其机制可能与miR-17表达减少从而介导Bax、Caspase-3的表达增加、Bcl-2的表达减少有关,FA可以直接上调miR-17的表达,减少Bax、Caspase-3,增加Bcl-2的表达,最终减少LPS诱发的PMVECs凋亡,提示一定浓度的FA可能是急性呼吸窘迫综合征治疗的有效药物。Objective To explore the expression of miR-17 in the apoptosis of pulmonary microvascular endothelial cells(PMVECs)as well as effect and mechanism of ferulic acid(FA)on lipopolysaccharide(LPS)-induced apoptosis of PMVECs.Methods This study was an experimental study,PMVECs were stimulated with FA at different doses(O mg/L,O.1 mg/L,1 mg/L,10 mg/L,100 mg/L)for 24h,the methyl tetrazolium(MTT)was used to detect the cell activity,and appropriate FA concentration was selected for subsequent experiments.PMVECs were divided into the control group,LPS group,low-dose FA group(LPS+0.1 mg/L FA),medium-dose FA group(LPS+1 mg/L FA)and high-dose FA group(LPS+10 mg/LFA),and the flow cytometry was used to analyze the apoptosis rate of cells.Caspase-3,Bax protein and Bcl-2 protein were detected by western blotting technique,and miR-17 was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PC)to determine the impacts of FA on the apoptosis of PMVECs.PMVECs were divided into the control group,LPS group,LPS+FA(LPS+10mg/LFA),LPS+FA+inhibitor NC group,LPS+FA+miR-17 inhibitor group.The apoptosis rate and expression of apoptotic protein and miR-17 of groups were detected to determine the effect targets of FA.Results The 1oo mg/L FA was inapplicable.Compared with the control group,the apoptosis rate,Caspase-3 and Bax of LPS group increased,and Bcl-2 protein and miR-17 decreased(P<0.05 at all points).Compared with LPS group,the apoptosis rate,Caspase-3 and Bax of low-dose FA group,medium-dose FA group and high-dose FA group decreased,Bcl-2 protein and miR-17 increased(P<0.05 at all points),and the apoptosis rate of high-dose FA group decreased compared with that of low-dose FA group(P<0.05).The expression of miR-17in the LPS+FA+miR-17 inhibitor group was lower than that in the LPS+FA+inhibitor NC group,the apoptosis rate,Caspase-3 and Bax were higher,while Bcl-2 protein was lower(P<0.05 at all points).Conclusions The inflammatory state induced by LPS can inhibit the expression of miR-17,thereby cause the apoptosis

关 键 词：细胞凋亡 肺微血管内皮细胞 微小RNA-17 阿魏酸 

分 类 号：R563.8[医药卫生—呼吸系统]