Revelation of the dynamic progression of hypoxia-reoxygenation injury by visualization of the lysosomal hydrogen peroxide.
Hydrogen peroxide (H2O2) plays an important role in pathological conditions, such as cerebral ischemia-reperfusion (I-R) injury. Fluorescent probes may serve as valuable tools to detect the amount, temporal and spatial distribution of H2O2in living cells. To investigate the role of lysosomal H2O2involved in cerebral I-R injury, we designed and synthesized a lysosome-targetable two-photon fluorescent probe ztl-4, through expansion and substitution of the original pyridazinone scaffold, conjugation of electronic-donating aromatic ring and precise terminal modification of the alkyl linker. The probe ztl-4 exhibited fast, sensitive and highly selective response toward H2O2. ztl-4 could image exogenous H2O2in SH-SY5Y cells and brain slices. In addition, ztl-4 was located in lysosomes with high colocalization coefficient compared with LysoTracker. ztl-4 was further applied for detecting the endogenous generation of H2O2in SH-SY5Y cells subjected to oxygen and glucose deprivation (OGD) or OGD/reoxygenation (OGD/R) injury. Both OGD- and OGD/R-induced cell injury caused a time-dependent increase of H2O2production within lysosomes. Moreover, OGD/R-treated cells showed much more amount of H2O2than OGD-treated cells, indicating that reoxygenation will promote H2O2accumulation in lysosomes of post-hypoxia cells. Therefore, the probe is suitable for monitoring the dynamic changes of lysosomal H2O2in cells.
Department of Molecular and Cellular Pharmacology, Peking University School of Pharmaceutical Sciences, Beijing, China.
Yuanjun
Y
Zhu
Department of Medicinal Chemistry, Peking University School of Pharmaceutical Sciences, Beijing, China.
Tongliang
T
Zhou
Department of Medicinal Chemistry, Peking University School of Pharmaceutical Sciences, Beijing, China.
Lingfei
L
Yang
Medical and Healthy Analysis Center, Peking University, Beijing, China.
Lan
L
Yuan
Department of Medicinal Chemistry, Peking University School of Pharmaceutical Sciences, Beijing, China. Electronic address: leiliang@bjmu.edu.cn.
Lei
L
Liang
Department of Medicinal Chemistry, Peking University School of Pharmaceutical Sciences, Beijing, China. Electronic address: pingxu@bjmu.edu.cn.
Ping
P
Xu
D000818:Animals; D002478:Cells, Cultured; D003199:Computer Systems; D018450:Disease Progression; D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide; D007700:Kinetics; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D015427:Reperfusion Injury / Q000378:metabolism* / Q000473:pathology*
D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide
Fluorescent probe; Hydrogen peroxide; Lysosome; Oxygen and glucose deprivation/reoxygenation