摘要
目的初步探索自组装短肽Sciobio-Ⅱ抑制氧化低密度脂蛋白(ox-LDL)诱导的脐静脉内皮细胞(HUVEC)凋亡的作用机制。方法构建自组装短肽体系,体外培养HUVEC,分为5组并给予含不同试剂的DMEM完全培养基培养:control组(PBS)、ox-LDL组(200 mg/L ox-LDL)、Sciobio-Ⅱ组(150 mg/L Sciobio-Ⅱ)及ALL组(200 mg/L ox-LDL+150 mg/L Sciobio-Ⅱ)、LY组[200 mg/L ox-LDL+150 mg/L Sciobio-Ⅱ+25μmol/L血管内皮生长因子(VEGF)/Notch信号通路抑制剂LY450139]。刚果红染色宏观定性观察短肽成胶能力;CCK-8试验检测各组细胞增殖;末端DNA转移酶dUTP缺口末端标记(TUNEL)、Annexin V/PI试验法检测细胞凋亡;实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blot)试验检测Bax、B淋巴细胞瘤-2(Bcl-2)、Notch1、血管内皮生长因子受体2(VEGFR-2)基因及蛋白的表达情况。结果刚果红染色结果表明Sciobio-Ⅱ可以自组装形成三维体系;细胞增殖结果显示:与control组比较,ox-LDL组的HUVEC增殖能力受抑制(P<0.05);与ox-LDL组比较,Sciobio-Ⅱ组、ALL组的HUVEC增殖能力提高,Sciobio-Ⅱ组高于ALL组(P<0.05)。细胞凋亡实验结果:与control组比较,ox-LDL组细胞凋亡率增加(P<0.05);与ox-LDL组比较,Sciobio-Ⅱ组和ALL组细胞凋亡率均降低,Sciobio-Ⅱ组低于ALL组(P<0.05);qRT-PCR及Western blot实验(SYBR法)结果表明:ox-LDL组Bcl-2、VEGFR-2基因与蛋白表达水平低于control组,ALL组高于ox-LDL组,LY组低于ALL组;ox-LDL组Bax、Notch1基因与蛋白表达水平高于control组,ALL组低于ox-LDL组,LY组高于ALL组(均P<0.05)。结论自组装短肽Sciobio-Ⅱ可通过激活VEGF/Notch信号通路减轻ox-LDL引起的HUVEC凋亡。
Objective To preliminarily investigate the action mechanism of self-assembling short peptide Sciobio-Ⅱin inhibiting the apoptosis of human umbilical vein endothelial cells(HUVEC)induced by oxidized low density lipoprotein(ox-LDL).Methods The self-assembling short peptides system was constructed.HUVEC were cultured in vitro,and were divided into five groups and given different reagents of DMEM complete medium:the control group(PBS),the ox-LDL group(200 mg/L of ox-LDL),Sciobio-Ⅱgroup(150 mg/L of Sciobio-Ⅱ)and ALL group(200 mg/L of ox-LDL+150 mg/L of Sciobio-Ⅱ),LY group(200 mg/L of ox-LDL+150 mg/L of Sciobio-Ⅱ+25μmol/L of signal pathway inhibitor LY450139).The Congo red staining was used to observe the gelatinizing ability of short peptides.The cell proliferation in each group was detected by the CCK-8 experiment.The cellular apoptosis was detected by TUNEL and Annexin V/PI experiment.The expression levels of Bax,Bcl-2,Notch1 and VEGFR-2 gene and protein were detected by qRT-PCR and Western blot.Results The Congo red staining results showed that Sciobio-Ⅱcould be self-assembled to form a three-dimensional system.The cell proliferation results showed that compared with the control group,the proliferation of HUVEC in the ox-LDL group was inhibited(P<0.05).Compared with the ox-LDL group,the HUVEC proliferation in the Sciobio-Ⅱgroup and the ALL group was increased,and the Sciobio-Ⅱgroup was higher than the ALL group(P<0.05).The cell apoptosis experiment results showed that compared with the control group,the apoptosis in the ox-LDL group was increased(P<0.05),compared with the ox-LDL group,the apoptosis in the Sciobio-Ⅱgroup and ALL group was decreased,the Sciobio-Ⅱgroup was lower than ALL group(P<0.05);the qRT-PCR and Western blot experiment results showed that the gene and protein expression levels of Bcl-2 and VEGFR-2 genes and proteins in the ox-LDL group were lower than those in the control group,the ALL group was higher than the ox-LDL group,the LY group was lower than the ALL group.The gene a
作者
龚心琰
朱凌波
郑娟庆
龚剑萍
GONG Xinyan;ZHU Lingbo;ZHENG Juanqing;GONG Jianping(Department of Cardiology,Yiwu Central Hospital,Yiwu,Zhejiang 322000,China)
出处
《重庆医学》
CAS
2022年第22期3786-3791,共6页
Chongqing medicine
基金
浙江省义乌市科技计划项目(19-3-05)
浙江省金华市科技计划项目(2019-4-136)。
关键词
自组装短肽
氧化低密度脂蛋白
人脐静脉内皮细胞
增殖
细胞凋亡
self-assembling peptides
oxidized low density lipoprotein
human umbilical vein endothelial cells
proliferation
apoptosis
