PMID- 32514052
OWN - NLM
STAT- MEDLINE
DCOM- 20210119
LR  - 20211204
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 10
IP  - 1
DP  - 2020 Jun 8
TI  - Cilostazol restores autophagy flux in bafilomycin A1-treated, cultured cortical 
      astrocytes through lysosomal reacidification: roles of PKA, zinc and 
      metallothionein 3.
PG  - 9175
LID - 10.1038/s41598-020-66292-3 [doi]
LID - 9175
AB  - Cilostazol, a phosphodiesterase 3 inhibitor, reduces the amyloid-beta (Abeta) burden 
      in mouse models of Alzheimer disease by as yet unidentified mechanisms. In the 
      present study, we examined the possibility that cilostazol ameliorates lysosomal 
      dysfunction. Astrocytes treated with bafilomycin A1 (BafA1) exhibited markedly 
      reduced DND-189 and acridine orange (AO) fluorescence, indicating reduced 
      lysosomal acidity. In both cases, BafA1-induced alkalization was reversed by 
      addition of cilostazol, dibutyryl cAMP or forskolin. All three agents 
      significantly increased free zinc levels in lysosomes, and addition of the zinc 
      chelator TPEN abrogated lysosomal reacidification. These treatments did not raise 
      free zinc levels or reverse BafA1-mediated lysosomal alkalization in 
      metallothionein 3 (Mt3)-null astrocytes, indicating that the increases in zinc in 
      astrocytes were derived mainly from Mt3. Lastly, in FITC-Abeta-treated astrocytes, 
      cilostazol reversed lysosomal alkalization, increased cathepsin D activity, and 
      reduced Abeta accumulation in astrocytes. Cilostazol also reduced mHtt aggregate 
      formation in GFP-mHttQ74-expressing astrocytes. Collectively, our results present 
      the novel finding that cAMP/PKA can overcome the v-ATPase blocking effect of 
      BafA1 in a zinc- and Mt3-dependent manner.
FAU - Kim, Ha Na
AU  - Kim HN
AD  - Neural Injury Lab, Biomedical Research Center, Asan Institute for Life Sciences, 
      Asan Medical Center, Seoul, Korea.
FAU - Seo, Bo-Ra
AU  - Seo BR
AD  - Neural Injury Lab, Biomedical Research Center, Asan Institute for Life Sciences, 
      Asan Medical Center, Seoul, Korea.
FAU - Kim, Hyunjin
AU  - Kim H
AUID- ORCID: 0000-0003-0264-4531
AD  - Department of Neurology, University of Ulsan College of Medicine, Seoul, Korea; 
      Department of Neurology, Asan Medical Center, University of Ulsan College of 
      Medicine, Seoul, Korea.
FAU - Koh, Jae-Young
AU  - Koh JY
AD  - Department of Neurology, University of Ulsan College of Medicine, Seoul, Korea; 
      Department of Neurology, Asan Medical Center, University of Ulsan College of 
      Medicine, Seoul, Korea. jkko@amc.seoul.kr.
AD  - Neural Injury Lab, Biomedical Research Center, Asan Institute for Life Sciences, 
      Asan Medical Center, Seoul, Korea. jkko@amc.seoul.kr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200608
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Amyloid beta-Peptides)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Macrolides)
RN  - 0 (Metallothionein 3)
RN  - 0 (Mt3 protein, mouse)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Phosphodiesterase 3 Inhibitors)
RN  - 88899-55-2 (bafilomycin A1)
RN  - E0399OZS9N (Cyclic AMP)
RN  - EC 3.4.23.5 (Cathepsin D)
RN  - EC 3.6.1.- (Adenosine Triphosphatases)
RN  - J41CSQ7QDS (Zinc)
RN  - N7Z035406B (Cilostazol)
SB  - IM
MH  - Adenosine Triphosphatases/antagonists & inhibitors
MH  - Amyloid beta-Peptides/metabolism
MH  - Animals
MH  - Astrocytes/*cytology
MH  - Autophagy
MH  - Cathepsin D/metabolism
MH  - Cells, Cultured
MH  - Cilostazol/*pharmacology
MH  - Cyclic AMP/*metabolism
MH  - Enzyme Inhibitors
MH  - Hydrogen-Ion Concentration/drug effects
MH  - Lysosomes/*metabolism
MH  - Macrolides/*pharmacology
MH  - Metallothionein 3
MH  - Mice
MH  - Nerve Tissue Proteins/*metabolism
MH  - Phosphodiesterase 3 Inhibitors/*pharmacology
MH  - Zinc/*metabolism
PMC - PMC7280249
COIS- The authors declare no competing interests.
EDAT- 2020/06/10 06:00
MHDA- 2021/01/20 06:00
PMCR- 2020/06/08
CRDT- 2020/06/10 06:00
PHST- 2020/01/22 00:00 [received]
PHST- 2020/05/18 00:00 [accepted]
PHST- 2020/06/10 06:00 [entrez]
PHST- 2020/06/10 06:00 [pubmed]
PHST- 2021/01/20 06:00 [medline]
PHST- 2020/06/08 00:00 [pmc-release]
AID - 10.1038/s41598-020-66292-3 [pii]
AID - 66292 [pii]
AID - 10.1038/s41598-020-66292-3 [doi]
PST - epublish
SO  - Sci Rep. 2020 Jun 8;10(1):9175. doi: 10.1038/s41598-020-66292-3.