Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe

doi:10.7150/thno.33867

	Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe
	Lei, Yu 1; Ren, Wenming 2; Wang, Cheng-Kun 1; Tao, Rong-Rong 1; Xiang, Huai-Jiang 1; Feng, Li-Li 3; Gao, Yin-Ping 1; Jiang, Quan 1; Li, Xin 1; Hu, Youhong 2
刊名	THERANOSTICS
	2019
卷号	9 期号:19 页码:5672-5680
关键词	Fluorescent imaging fluorescent probe autophagy endothelial injury
ISSN号	1838-7640
DOI	10.7150/thno.33867
通讯作者	Li, Xin(lixin81@zju.edu.cn) ; Hu, Youhong(yhhu@simm.ac.cn) ; Han, Feng(fenghan169@njmu.edu.cn)
英文摘要	Autophagy is a dynamic and complicated catabolic process. Imaging autophagic flux can clearly advance knowledge of its pathophysiology significance. While the most common way autophagy is imaged relies on fluorescent protein-based probes, this method requires substantial genetic manipulation that severely restricts the application. Small fluorescent probes capable of tracking autophagic flux with good spatiotemporal resolution are highly demanable. Methods: In this study, we developed a small-molecule fluorogenic probe (AFG-1) that facilitates real-time imaging of autophagic flux in both intact cells and live mice. AFG-1 is inspired by the cascading nitrosative and acidic microenvironments evolving during autophagy. It operates over two sequential steps. In the first step, AFG-1 responds to the up-regulated peroxynitrite at the initiation of autophagy by its diphenylamino group being oxidatively dearylated to yield a daughter probe. In the second step, the daughter probe responds to the acidic autolysosomes at the late stage of autophagy by being protonated. Results: This pathway-dependent mechanism has been confirmed first by sequentially sensing ONOO- and acid in aqueous solution, and then by imaging autophagic flux in live cells. Furthermore, AFG-1 has been successfully applied to visualize autophagic flux in real-time in live mice following brain ischemic injury, justifying its robustness. Conclusion: Due to the specificity, easy operation, and the dynamic information yielded, AFG-1 should serve as a potential tool to explore the roles of autophagy under various pathological settings.
资助项目	National Key Research and Development Program of China[2016YFE0125400] ; National Natural Science Foundations of China[81473202] ; National Natural Science Foundations of China[81573411] ; National Natural Science Foundations of China[81225022] ; National Natural Science Foundations of China[21642007] ; National Natural Science Foundations of China[21778048] ; Zhejiang Provincial Natural Science Foundation of China[Z16H310003] ; Zhejiang Provincial Natural Science Foundation of China[R18H300001] ; National Science & Technology Major Project Key New Drug Creation and Manufacturing Program[2018ZX09711002]
WOS关键词	PEROXYNITRITE ; FLUORESCENT ; DESIGN ; CELLS
WOS研究方向	Research & Experimental Medicine
语种	英语
出版者	IVYSPRING INT PUBL
WOS记录号	WOS:000481600100017
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/289008]
专题	中国科学院上海药物研究所
通讯作者	Li, Xin; Hu, Youhong; Han, Feng
作者单位	1.Zhejiang Univ, Coll Pharmaceut Sci, Hangzhou 310058, Zhejiang, Peoples R China 2.Chinese Acad Sci, State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 3.Nanjing Med Univ, Sch Pharm, Nanjing 211166, Jiangsu, Peoples R China
推荐引用方式 GB/T 7714	Lei, Yu,Ren, Wenming,Wang, Cheng-Kun,et al. Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe[J]. THERANOSTICS,2019,9(19):5672-5680.
APA	Lei, Yu.,Ren, Wenming.,Wang, Cheng-Kun.,Tao, Rong-Rong.,Xiang, Huai-Jiang.,...&Han, Feng.(2019).Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe.THERANOSTICS,9(19),5672-5680.
MLA	Lei, Yu,et al."Visualizing Autophagic Flux during Endothelial Injury with a Pathway-Inspired Tandem-Reaction Based Fluorogenic Probe".THERANOSTICS 9.19(2019):5672-5680.