1.Effect of mulberry anthocyanin suppressing autophagy on morphology and function of vascular endothelial cells through activating PI3K/Akt/mTOR pathway
Kun WANG ; Chuancai DAN ; Xiang HU ; Zhaoyang PI ; Huanlu WANG ; Dandan ZHU ; Yumeng ZHOU ; Shaofeng XIONG
Chongqing Medicine 2025;54(2):289-296
Objective To investigate the effect of mulberry anthocyanin on the morphology and func-tion 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 inju-ry model group.10,25,50 μmol/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 ob-served.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 af-ter 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 in-hibit 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.

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