摘要
目的 本研究拟通过体外实验研究槲皮素(Quercetin,QCT)与AC16心肌细胞增殖的关系,探析QCT阻止AC16心肌细胞增殖作用与Wnt/β-catenin信号通路的关系。方法 选用不同浓度QCT对AC16细胞进行干预,使用倒置显微镜拍照观察法、台盼蓝计数法和CCK-8法检测QCT对AC16细胞增殖的影响。Western blot检测QCT对磷酸化-β-连环蛋白(Phosphorylated β-catenin,p-β-catenin)、细胞性骨髓瘤样癌基因(c-Myc)、β-连环蛋白(β-catenin)、低密度脂蛋白受体相关蛋白5(Low density lipoprotein receptor-related protein 5,LRP5)和低密度脂蛋白受体相关蛋白6(Low density lipoprotein receptor-related protein 6,LRP6)表达的影响。台盼蓝计数法检测过表达β-catenin ΔN、LRP5和LRP6基因后研究QCT对细胞增殖的影响以及沉默LRP5和LRP6基因后研究QCT对细胞增殖的影响。结果 与对照组比,QCT使得AC16细胞活细胞数下降,抑制增殖率,且具有浓度依赖性。条件摸索显示10和20μmol/L的QCT会导致AC16细胞中p-β-catenin蛋白修饰水平显著增加(P<0.05),c-Myc、β-catenin、LRP5和LRP6蛋白表达水平显著下调(P<0.05),因此后续实验选择10μmol/L的QCT作为干预浓度。而AC16细胞中过表达β-catenin ΔN、LRP5和LRP6基因,与单独给药组相比,过表达相关基因后10μmol/L的QCT引起的细胞增殖抑制现象被有效逆转(P<0.05)。另外,沉默LRP5和LRP6再联合10μmol/L的QCT干预,AC16细胞的增殖抑制并未产生协同现象(P>0.05)。结论 QCT可以通过下调LRP5/6表达以抑制Wnt/β-catenin信号通路从而发挥其阻止AC16心肌细胞增殖的作用。
Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylatedβ-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressingβ-cateninΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20µmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10µmol/L QCT was chosen as the intervention concentration.Overexpression ofβ-cateninΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10µmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10µmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.
作者
钟小妹
沈芳
沃达
彭军
朱伟东
任丹妮
ZHONG Xiaomei;SHEN Fang;WO Da;PENG Jun;ZHU Weidong;REN Danni(College of Integrative Medicine,Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fujian Key Laboratory of Integrative Medicine on Geriatric,Fuzhou,350122,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第3期643-651,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
福建中医药大学青年科技创新培育计划项目(XQB202201),负责人:任丹妮
福建中医药大学高层次人才科研启动资金项目(X2021001-人才):活心丸通过Wnt信号通路对心脏发挥保护作用的机制研究,负责人:任丹妮
福建中医药大学高层次人才科研启动资金项目(X2019001-人才):LRP5/6通过维持GPCR的稳态对心脏发挥保护作用的机制研,负责人:任丹妮。
