PMID- 34060443
OWN - NLM
STAT- MEDLINE
DCOM- 20210602
LR  - 20210602
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 37
IP  - 6
DP  - 2021 Jun
TI  - [Establishment and evaluation of hepatocyte injury model induced by 
      LPS/D-galactosamine in vitro].
PG  - 495-500
AB  - Objective To establish a novel hepatocyte injury model induced by 
      lipopolysaccharide/D-galactosamine (LPS/D-GalN) in vitro. Methods Freshly 
      isolated mouse primary hepatocytes were cultured in vitro and treated with 
      different doses of tumor necrosis factor-alpha (TNF-alpha) and 5 mg/mL of D-GalN. The 
      supernatants from hepatocyte culture were detected for alanine aminotransferase 
      (ALT) activity by chemiluminescence assay. Bone marrow-derived macrophages 
      (BMDMs) were stimulated with 1 mug/mL of LPS and the level of TNF-alpha in 
      supernatants were detected by ELISA. Primary hepatocytes were treated with the 
      BMDM supernatants combined with 5 mg/mL D-GalN or 50 ng/mL actinomycin D (ActD) 
      for 24 hours. The level of ALT from hepatocyte supernatant was detected and 
      morphology of hepatocytes was observed with microscopy. BMDMs and hepatocytes 
      were co-cultured and treated with 1 mug/mL of LPS combined with D-GalN or ActD for 
      24 hours. Hepatocyte injury was reflected by the ALT activity and hepatocyte 
      morphology. Results The ALT activity was significantly increased in the 
      supernatants of hepatocytes treated with TNF-alpha and D-GalN, indicating the obvious 
      hepatocyte injury. Co-treatment with LPS-primed BMDM supernatants and D-GalN or 
      ActD could cause hepatocyte injury, as reflected by markedly increased ALT 
      activity and the deformed and cracked hepatocytes. In the context of co-culture 
      of BMDM and hepatocytes, treatment with LPS and D-GalN led to obvious hepatocyte 
      injury as expected. LPS combined with ActD could not cause hepatocyte injury, 
      since the BMDMs started to die earlier than they could secret TNF-alpha to destruct 
      hepatocytes. Hepatocytes with normal morphology and deformed BMDMs were observed. 
      Conclusion LPS/D-GalN can be used to induce hepatocyte injury in vitro. D-GalN, 
      rather than ActD, should be used as a transcriptional inhibitor when the TNF-alpha 
      -induced hepatocyte injury is evaluated in a co-culture system of BMDMs and 
      hepatocytes.
FAU - He, Jia
AU  - He J
AD  - Henan University Joint National Laboratory for Antibody Drug Engineering, Kaifeng 
      475004, China.
FAU - Liu, Genyu
AU  - Liu G
AD  - Institute of Military Cognition and Brain Science, Academy of Military Medical 
      Sciences, Beijing 100850, China.
FAU - Zhang, Yaolin
AU  - Zhang Y
AD  - Institute of Military Cognition and Brain Science, Academy of Military Medical 
      Sciences, Beijing 100850, China.
FAU - Zhang, Jun
AU  - Zhang J
AD  - Henan University Joint National Laboratory for Antibody Drug Engineering, Kaifeng 
      475004, China.
FAU - Wang, Qingyang
AU  - Wang Q
AD  - Institute of Military Cognition and Brain Science, Academy of Military Medical 
      Sciences, Beijing 100850, China. *Corresponding author, E-mail: 
      tansun0532@163.com.
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 7535-00-4 (Galactosamine)
RN  - EC 2.6.1.2 (Alanine Transaminase)
SB  - IM
MH  - Alanine Transaminase
MH  - Animals
MH  - *Galactosamine/toxicity
MH  - Hepatocytes
MH  - *Lipopolysaccharides
MH  - Mice
MH  - Tumor Necrosis Factor-alpha
EDAT- 2021/06/02 06:00
MHDA- 2021/06/03 06:00
CRDT- 2021/06/01 08:43
PHST- 2021/06/01 08:43 [entrez]
PHST- 2021/06/02 06:00 [pubmed]
PHST- 2021/06/03 06:00 [medline]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Jun;37(6):495-500.