[1]石颖,陈子扬,陈凯铭,等.补阳还五汤对高糖环境下miR-21介导的人视网膜微血管内皮细胞自噬的影响[J].眼科新进展,2022,42(11):858-862.[doi:10.13389/j.cnki.rao.2022.0177]
　SHI Ying,CHEN Ziyang,CHEN Kaiming,et al.Effect of Buyang Huanwu decoction on miR-21-mediated autophagy in human retinal capillary endothelial cells under high glucose[J].Recent Advances in Ophthalmology,2022,42(11):858-862.[doi:10.13389/j.cnki.rao.2022.0177]
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补阳还五汤对高糖环境下miR-21介导的人视网膜微血管内皮细胞自噬的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
42卷
期数:
2022年11期
页码:
858-862
栏目:
实验研究
出版日期:
2022-11-05

文章信息/Info

Title:
Effect of Buyang Huanwu decoction on miR-21-mediated autophagy in human retinal capillary endothelial cells under high glucose
作者:
石颖; 陈子扬; 陈凯铭; 胡艳红; 胡俊; 陈胜
350122　福建省福州市，福建中医药大学（石颖，陈凯铭）；350003　福建省福州市，福建省中医药科学院综合门诊部（陈子扬）；350003　福建省福州市，福建中医药大学附属第二人民医院眼科（胡艳红，胡俊，陈胜）
Author(s):
SHI Ying1; CHEN Ziyang2; CHEN Kaiming1; HU Yanhong3; HU Jun3; CHEN Sheng3
1.Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian Province,China
2.Comprehensive Outpatient Department of Fujian Academy of Chinese Medicine,Fuzhou 350003,Fujian Province,China
3.The Second Affiliated Hospital of Fujian Traditional Chinese Medical University,Fuzhou 350003,Fujian Province,China
关键词:
补阳还五汤; miR-21; 自噬; 高糖; 视网膜微血管内皮细胞
Keywords:
Buyang Huanwu decoction;  miR-21;  autophagy;  high glucose;  retinal capillary endothelial cells
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2022.0177
文献标志码:
A
摘要:
目的　观察补阳还五汤对高糖环境下miR-21介导的人视网膜微血管内皮细胞（HRCECs）自噬的影响，以期为DR自噬调控机制及药物治疗提供依据。方法　HRCECs随机分5组，即正常对照组、高糖组、补阳还五汤组、miR-21 mimic组、miR-21 mimic+补阳还五汤组。除正常对照组外，其余各组细胞于25.0 g·L-1葡萄糖环境中培养,建立高糖损伤模型，补阳还五汤组加用5 g·L-1补阳还五汤水提液干预；miR-21 mimic组孵育转染 miR-21 mimic的HRCECs；miR-21 mimic+补阳还五汤组用5 g·L-1补阳还五汤水提液干预转染 miR-21 mimic的HRCECs。荧光显微镜下观察转染效率，实时荧光定量PCR检测各组细胞中miR-21的表达，Western blot检测各组细胞中LC3蛋白表达。结果　经转染miR-21 mimic后，大部分HRCECs呈绿色荧光，荧光强度为32.708±1.001。实时荧光定量PCR检测结果显示，miR-21 mimic组HRCECs的miR-21相对表达量为106.677±5.787，明显高于高糖组（5.892±0.341）和正常对照组（0.993±0.015），差异均有统计学意义（均为P<0.05）。高糖组和miR-21 mimic组HRCECs中LC3蛋白相对表达量均较正常对照组升高，差异均有统计学意义（均为P<0.05），且miR-21 mimic组HRCECs中LC3蛋白相对表达量高于高糖组（P=0.001）。补阳还五汤组HRCECs中LC3蛋白相对表达量低于高糖组和miR-21 mimic组（均为P<0.05），与正常对照组比较差异无统计学意义（P=0.197）。miR-21 mimic+补阳还五汤组HRCECs中LC3蛋白相对表达量低于miR-21 mimic组，但高于补阳还五汤组，差异均有统计学意义（均为P<0.05），与高糖组比较差异无统计学意义（P=0.930）。结论　高糖环境下，miR-21介导HRCECs的自噬，补阳还五汤可以下调HRCECs的miR-21表达，进而减少细胞自噬。
Abstract:
Objective　To investigate the effect of Buyang Huanwu decoction (BHD) on miR-21-mediated autophagy in human retinal capillary endothelial cells (HRCECs) under high glucose (HG) to provide a basis for research on the DR autophagy regulation mechanism and drug therapy. Methods　HRCECs were randomly divided into five groups: normal control (NC) group, HG group, BHD group, miR-21 mimic group, and miR-21 mimic+BHD group. Cells were cultured in 25 g·L-1 glucose and HG damage models were established except the NC group. In addition, cells in the BHD group were intervened with 5 g·L-1 BHD water extract; cells in the miR-21 mimic group were transfected with miR-21 mimic; cells in the miR-21 mimic+BHD group were transfected with miR-21 mimic and then were intervened with 5 g·L-1 BHD water extract. Transfection efficiency was assessed under the fluorescence microscope. The miR-21 expression in cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The LC3 protein expression was measured by Western blot. Results　After transfection with miR-21 mimic, most HRCECs showed green fluorescence, with an average fluorescence intensity of 32.708±1.001. The qRT-PCR results showed that the relative expression of miR-21 in the miR-21 mimic group was 106.677±5.787, significantly higher than that in the HG group (5.892±0.341) and NC group (0.993±0.015) (both P<0.05). LC3 protein expression rose in both the HG and miR-21 mimic groups compared with the NC group (both P<0.05), and LC3 protein expression in the miR-21 mimic group was higher than that in the HG group (P=0.001). LC3 protein expression in the BHD group was lower than that in the HG and miR-21 mimic groups (both P<0.05), but showed no significant difference compared with the NC group (P=0.197). LC3 protein expression in the miR-21 mimic+BHD group was lower than that in the miR-21 mimic group, but higher than that in the BHD group (both P<0.05). There was no significant difference in LC3 protein expression between the miR-21 mimic+BHD group and the HG group (P=0.930). Conclusion　miR-21 mediates the autophagy of HRCECs under HG. BHD can downregulate the miR-21 expression in HRCECs and then reduce autophagy.

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备注/Memo

备注/Memo:
福建省卫健委科技计划项目（编号：2020CXB034）；福建省自然科学基金计划项目（编号：2022J01834）
更新日期/Last Update: 2022-11-05