PMID- 31180558
OWN - NLM
STAT- MEDLINE
DCOM- 20191216
LR  - 20211204
IS  - 1791-3004 (Electronic)
IS  - 1791-2997 (Linking)
VI  - 20
IP  - 1
DP  - 2019 Jul
TI  - Bupivacaine at clinically relevant concentrations induces toxicity in human 
      intervertebral disc cells via the induction of autophagy in vitro.
PG  - 837-843
LID - 10.3892/mmr.2019.10279 [doi]
AB  - It has been reported that bupivacaine, the most widely used local anesthetic to 
      relieve discogenic back pain in clinical settings, is cytotoxic to intervertebral 
      disc (IVD) cells in vitro; however, the precise mechanisms of cytotoxicity 
      induced by bupivacaine remain unclear. Autophagy is an intracellular lysosomal 
      degradation process that is important for cellular survival. The present study 
      investigated the role of autophagy in the survival of IVD cells subjected to 
      bupivacaine treatment. Human nucleus pulposus (NP) cells isolated from IVD cells 
      were exposed to various concentrations of bupivacaine for 2, 6 and 12 h, and 
      analyzed for cellular viability using MTT assay and western blotting. 
      Additionally, autophagosome formation and autophagy‑associated biomarkers were 
      evaluated by electron microscopy and western blotting to determine the autophagic 
      activity and signaling alterations in NP cells under bupivacaine treatment. 
      Furthermore, autophagic activity was inhibited in vitro using 3‑methyladenine to 
      further analyze the association between autophagy and apoptosis in 
      bupivacaine‑treated NP cells. Bupivacaine exhibited time‑ and dose‑dependent 
      cytotoxic effects on human IVD cells at clinically relevant concentrations. 
      Bupivacaine increased autophagic activity by promoting autophagosome formation, 
      and LC3‑II and Beclin‑1 production. Additionally, bupivacaine inhibited protein 
      kinase B (Akt)/mammalian target of rapamycin (mTOR)/S6 kinase (S6K) signaling, 
      which is a negative regulator of autophagic activity. Of note, pharmacological 
      inhibition of autophagy alleviated bupivacaine‑induced cytotoxicity of IVD cells. 
      The findings indicated that application of clinically relevant concentrations of 
      bupivacaine upregulated autophagic activity via inhibition of Akt/mTOR/S6K 
      signaling. In addition, the inhibition of autophagic activation served as a 
      protective mechanism against bupivacaine‑induced cytotoxicity. Collectively, 
      these findings may provide novel insight into the mechanisms underlying 
      cytotoxicity induced by bupivacaine when controlling spine‑associated pain.
FAU - Yang, Ge
AU  - Yang G
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
FAU - Li, Zhuoyang
AU  - Li Z
AD  - Department of Orthopedics, First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China.
FAU - Mei, Haibo
AU  - Mei H
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
FAU - Ye, Weihua
AU  - Ye W
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
FAU - Huang, Shengxiang
AU  - Huang S
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
FAU - Liu, Kun
AU  - Liu K
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
FAU - Tan, Qian
AU  - Tan Q
AD  - Department of Orthopedics, Hunan Children's Hospital, The Pediatric Academy of 
      University of South China, Changsha, Hunan 410007, P.R. China.
LA  - eng
PT  - Journal Article
DEP - 20190522
PL  - Greece
TA  - Mol Med Rep
JT  - Molecular medicine reports
JID - 101475259
RN  - 0 (Anesthetics, Local)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.1 (Ribosomal Protein S6 Kinases)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - Y8335394RO (Bupivacaine)
SB  - IM
MH  - Adult
MH  - Anesthetics, Local/*toxicity
MH  - Autophagy/*drug effects
MH  - Bupivacaine/*toxicity
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Dose-Response Relationship, Drug
MH  - Female
MH  - Humans
MH  - Intervertebral Disc/cytology/*drug effects/pathology
MH  - Male
MH  - Middle Aged
MH  - Nucleus Pulposus/cytology/*drug effects/pathology
MH  - Proto-Oncogene Proteins c-akt/metabolism
MH  - Ribosomal Protein S6 Kinases/metabolism
MH  - Signal Transduction/drug effects
MH  - TOR Serine-Threonine Kinases/metabolism
EDAT- 2019/06/11 06:00
MHDA- 2019/12/18 06:00
CRDT- 2019/06/11 06:00
PHST- 2018/08/20 00:00 [received]
PHST- 2019/04/17 00:00 [accepted]
PHST- 2019/06/11 06:00 [pubmed]
PHST- 2019/12/18 06:00 [medline]
PHST- 2019/06/11 06:00 [entrez]
AID - 10.3892/mmr.2019.10279 [doi]
PST - ppublish
SO  - Mol Med Rep. 2019 Jul;20(1):837-843. doi: 10.3892/mmr.2019.10279. Epub 2019 May 
      22.