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1.400038 重庆,陆军军医大学(第三军医大学)军事预防医学系: 军队流行病学教研室
2.400038 重庆,陆军军医大学(第三军医大学)军事预防医学系: 营养与食品卫生学教研室
[李亚斐,E-mail: liyafei2008@hotmail.com LI Yafei, E-mail: liyafei2008@hotmail.com
收稿:2021-07-26,
修回:2021-11-04,
纸质出版:2022-02-28
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胡琴, 申卉, 张婷, 等. 二氢杨梅素通过Prohibitin 2增强血管内皮细胞线粒体自噬[J]. 陆军军医大学学报, 2022,44(4):356-362.
Qin HU, Hui SHEN, Ting ZHANG, et al. Dihydromyricetin induces mitophagy by regulating Prohibitin 2 pathway in vascular endothelial cells[J]. Journal of Army Medical University, 2022, 44(4): 356-362.
胡琴, 申卉, 张婷, 等. 二氢杨梅素通过Prohibitin 2增强血管内皮细胞线粒体自噬[J]. 陆军军医大学学报, 2022,44(4):356-362. DOI: 10.16016/j.2097-0927.202107159.
Qin HU, Hui SHEN, Ting ZHANG, et al. Dihydromyricetin induces mitophagy by regulating Prohibitin 2 pathway in vascular endothelial cells[J]. Journal of Army Medical University, 2022, 44(4): 356-362. DOI: 10.16016/j.2097-0927.202107159.
目的
2
观察二氢杨梅素对血管内皮细胞线粒体自噬的影响,并探讨Prohibitin 2在其中的作用。
方法
2
用不同浓度(0、0.1、1、10、100 μmol/L)二氢杨梅素处理人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)12 h后,流式细胞术和荧光显微镜观测MitoTracker
®
Deep Red (MTDR)荧光强度,RT-PCR和Western blot分别检测Prohibitin 2 mRNA及其蛋白表达。用氯喹预处理HUVECs 1 h后,再用不同浓度二氢杨梅素处理12 h或10 μmol/L羰基氰化物间氯苯腙(线粒体自噬激动剂)处理6 h,流式细胞术检测线粒体自噬。另用control siRNA或Prohibitin 2 siRNA转染HUVECs后,再用二氢杨梅素处理12 h,流式细胞术检测细胞线粒体自噬。
结果
2
与无二氢杨梅素(0 μmol/L)处理相比,≥0.1 μmol/L二氢杨梅素处理显著降低血管内皮细胞MTDR荧光强度;同时,二氢杨梅素处理可明显增加Prohibitin 2 mRNA和蛋白表达水平(
P
<0.05);当以氯喹预处理抑制溶酶体功能后,与无二氢杨梅素处理相比,≥10 μmol/L二氢杨梅素处理可以显著增加血管内皮细胞线粒体自噬,与羰基氰化物间氯苯腙效应相一致(
P
<0.05)。与control siRNA处理组相比,以siRNA转染沉默Prohibitin 2后二氢杨梅素对线粒体自噬的诱导作用被明显抑制(
P
<0.05)。
结论
2
二氢杨梅素可通过调控Prohibitin 2增强血管内皮细胞线粒体自噬。
Objective
2
To observe the effects of dihydromyricetin on mitophagy in vascular endothelial cells
and to clarify the underlying role of Prohibitin 2 in the process.
Methods
2
After human umbilical vein endothelial cells (HUVECs) were treated with different concentrations of dihydromyricetin (0
0.1
1
10
100 μmol/L) for 12 h
intensity of MitoTracker
®
Deep Red (MTDR
a widely used mitochondria-selective probe) was determined for mitochondrial staining by fluorescence microscopy and flow cytometry
and the expression of Prohibitin 2 at mRNA and protein levels as determined by qRT-PCR
and Western blotting
respectively. Furthermore
after treatment with of without control/Prohibitin 2 siRNA
HUVECs were incubated with dihydromyricetin for 12 h or carbonyl cyanide 3-chlorophenyl hydrazone (CCCP) for 6 h in the presence of the lysosomal blocker chloroquine
and then subjected to flow cytometry analysis to determine mitophagy.
Results
2
As compared with the non-dihydromyricetin group
dihydromyricetin (≥ 0.1μmol/L) treatment markedly decreased MTDR fluorescence levels (
P
<
0.05)
and meanwhile
the treatment significantly increased the mRNA and protein levels of Prohibitin 2 (
P
<
0.05). After pretreated with chloroquine
dihydromyricetin (≥10 μmol/L) significantly enhanced mitophagy in HUVECs
which was consistent with the effect of CCCP (
P
<
0.05). After Prohibitin 2 siRNA transfection
mitophagy induced by dihydromyricetin was significantly suppressed when compared with the control siRNA treatment group (
P
<
0.05).
Conclusion
2
Dihydromyricetin enhances mitophagy in vascular endothelial cells by regulating Prohibitin 2.
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