摘要
目的 观察桑葚花色苷对血管内皮细胞(VEC)形态、功能的影响,并在自噬水平探讨其可能的机制。方法 体外培养VEC,分为5.5/0 mmol/L(对照组)及11/0.125、22/0.250、33/0.500、44/1.000、55/2.000 mmol/L的高糖/高脂组,测定各组细胞功能水平和自噬水平,选择最佳高糖/高脂浓度作为VEC功能受损的模型组。在模型组中加入10、25、50μmol/L桑葚花色苷进行处理,分为低、中、高浓度桑葚花色苷组,观察桑葚花色苷对高糖/高脂应激下的VEC形态、功能和自噬的影响,接着加入自噬调节剂(雷帕霉素)研究其作用机制。HE染色观察细胞数量和形态,CCK-8法检测VEC活力,流式细胞仪检测VEC活性氧(ROS)水平,Western blot法检测VEC内Beclin-1和p62蛋白表达水平,试剂盒法检测VEC内一氧化氮(NO)和内皮素-1(ET-1)水平。结果 与对照组比较,中、高浓度的高糖/高脂组细胞数量和形态发生改变,细胞活力、p62、NO水平明显降低,ROS、Beclin-1、ET-1水平明显升高(P<0.05)。与模型组比较,桑葚花色苷组细胞数量增加、形态得到改善,细胞活力、p62、NO水平明显升高,ROS、Beclin-1、ET-1水平明显降低(P<0.05)。与高浓度桑葚花色苷组比较,加入雷帕霉素后细胞数量减少,形态变得不规则,NO水平降低,ET-1水平升高(P<0.05)。与对照组比较,模型组p-磷脂酰肌醇-3-激酶(PI3K)、p-蛋白激酶B(Akt)相对表达水平明显降低(P<0.05);与模型组比较,高浓度桑葚花色苷组p-PI3K、p-Akt相对表达水平明显升高(P<0.05)。结论 桑葚花色苷通过激活PI3K/Akt/哺乳动物雷帕霉素靶蛋白(mTOR)通路抑制自噬,改善高糖/高脂应激下VEC的形态和功能。
Objective To investigate the effect of mulberry anthocyanin on the morphology and function of vascular endothelial cells(VEC)and its possible mechanism at the level of autophagy.Methods VEC were cultured in vitro and divided into 5.5/0 mmol/L control group,11/0.125,22/0.250,33/0.500,44/1.000,55/2.000 mmol/L high glucose/high fat groups.The cell functional levels and autophage levels in each group were detected.The optimal high glucose/high lipid concentration was selected as the VEC function injury model group.10,25,50μmol/L mulberry anthocyanins were added into the model group for processing and divided into the low,middle and high concentrations of mulberry anthocyanin groups.The effects of mulberry anthocyanin on the morphology,function and autophagy of VEC in high sugar and high fat stress were observed.Then the autophagy regulator(rapamycin)was added to study its action mechanism.HE staining was used to observe the number and morphology of cells,the CCK-8 assay was used to detect the viability of VEC,the flow cytometry was used to detect the level of reactive oxygen species(ROS)in VEC,the Western blot was used to detect the expression level of Beclin-1 and p62 proteins in VEC,and the kit method was used to detect the level of nitric oxide(NO)and endothelin-1(ET-1)in VEC.Results Compared with the control group,the number and morphology of cells in the middle and high concentrations of high glucose/high fat groups were changed,the cell viability,p62,NO levels were significantly decreased(P<0.05),while the ROS,ET-1,and Beclin-1 expression levels were increased significantly(P<0.05).Compared with the model group,the number of cells in the mulberry anthocyanin groups was increased,morphology was improved,the cellular viability,p62 and No levels were significantly increased,and ROS,Beclin-1 and ET-1 levels were significantly decreased(P<0.05).Compared with the high concentration mulberry anthocyanin group,the cells number was decreased after adding rapamycin,the morphology changed to be irregular,the NO level was decreased,and the ET level was increased(P<0.05).Compared with the control group,the p-phosphatidylinositol 3-kinase(PI3K)and p-protein kinase B(Akt)relative expression levels in the model group were significantly decreased(P<0.05);compared with the model group,the p-PI3K and p-Akt relative expression levels in the high concentration mulberry anthocyanin group were significantly increased(P<0.05).Conclusion Mulberry anthocyanin could inhibit autophagy by activating the PI3K/Akt/mammalian target of rapamycin(mTOR)signaling pathway and improve the morphology and function of VEC under high glucose and lipid stress.
作者
王昆
但传才
胡翔
皮朝阳
王浣璐
朱丹丹
周予梦
熊绍风
WANG Kun;DAN Chuancai;HU Xiang;PI Zhaoyang;WANG Huanlu;ZHU Dandan;ZHOU Yumeng;XIONG Shaofeng(Department of Pharmacy,First Affiliated Hospital of Nanchang University,Nanchang,Jiangxi 330000,China;Department of General Medicine,First Affiliated Hospital of Nanchang University,Nanchang,Jiangxi 330000,China;Jiangxi Provincial Tumor Hospital,Nanchang,Jiangxi 330029,China)
出处
《重庆医学》
2025年第2期289-296,共8页
Chongqing Medical Journal
基金
江西省中医药管理局科技计划项目(SZYYB20216461)。
作者简介
通信作者:熊绍风,E-mail:ndyfy07255@ncu.edu.cn。
