PMID- 24854919
OWN - NLM
STAT- MEDLINE
DCOM- 20150205
LR  - 20181202
IS  - 0376-2491 (Print)
IS  - 0376-2491 (Linking)
VI  - 94
IP  - 12
DP  - 2014 Apr 1
TI  - [Effects of lipopolysaccharide pretreatment on endotoxin tolerance of human 
      umbilical cord mesenchymal stem cells].
PG  - 948-51
AB  - OBJECTIVE: To explore the effects of lipopolysaccharide (LPS) pretreatment on 
      endotoxin tolerance of human umbilical cord mesenchymal stem cells (hUCMSCs) and 
      its possible mechanism. METHODS: hUCMSCs (1x10(4) cells/well) were exposed to 0, 
      0.1, 1.0, 10.0, 20.0, 30.0, 40.0, 50.0 microg/ml LPS for 24 h respectively. And the 
      cell viability of hUCMSCs was detected by 3-(4, 5-dimethylthiazol-2-yl)-2, 
      5-diphenyltetrazolium bromide (MTT). 1 microg/ml and 50.0 microg/ml LPS were used as 
      pretreatment and apoptosis induction concentrations respectively. Pyrrolidine 
      dithiocarbamate (PDTC) (20 micromol/L, pretreatment for 20 min) was used as a 
      specific inhibitor of nuclear transcription factor NF-kappaB. hUCMSCs were randomly 
      divided by Stata software into 7 groups: control (A), LPS induction (B), 
      pretreatment + LPS induction (C), PDTC (D), PDTC+ pretreatment + LPS induction 
      (E), pretreatment (F) and PDTC + pretreatment (G). The apoptosis of hUCMSCs was 
      measured by Hoechst 33258 staining and flow cytometry (FCM). The expressions of 
      NF-kappaB p65 and cellular FLICE-inhibitory protein (c-FLIP) were measured by Western 
      blot. RESULTS: The cell viability of 0, 0.1, 1.0, 10.0, 20.0, 30.0, 40.0, 50.0 
      microg/ml LPS groups were 100%, (117.0 +/- 8.8)%, (134.7 +/- 6.9)%, (105.3 +/- 8.3)%, (99.2 
      +/- 8.3)%, (84.2 +/- 9.3)%, (66.4 +/- 6.6)% and (59.2 +/- 8.0)% respectively. In 
      comparison with 0 microg/ml LPS group, the cell viability of 1.0 microg/ml LPS group 
      increased significantly (P = 0.004) while decreased in 40 and 50 microg/ml LPS groups 
      (P = 0.005, 0.002). Hoechst 33258 staining indicated that chromatin of hUCMSCs 
      was distributed evenly in group A; the apoptotic cell in group B dramatically 
      increased; and the apoptotic cell in group C significantly decreased in 
      comparison with that in group B. Apoptotic rates of groups A, B, C, D and E were 
      (2.8 +/- 0.8)%, (29.7 +/- 3.4)%, (17.8 +/- 3.0)%, (2.9 +/- 0.4)% and (23.2 +/- 2.6)% 
      respectively. Compared with group A, apoptosis rate significantly increased in 
      group B (P < 0.001). The apoptotic rate in group C significantly decreased than 
      that in group B (P < 0.001) while group E was higher than group C (P = 0.015). 
      The levels of NF-kappaB p65 and c-FLIP in group F (0.851 +/- 0.031, 0.534 +/- 0.053) was 
      higher than that in group A (0.220 +/- 0.021, 0.049 +/- 0.009) (both P < 0.001), G 
      (0.418 +/- 0.007, 0.299 +/- 0.061) (P < 0.001, P = 0.007). CONCLUSIONS: LPS 
      pretreatment can resist LPS-induced hUCMSCs apoptosis and enhance the ability of 
      endotoxin tolerance. And the mechanism may be related with activating the NF-kappaB 
      signaling pathway and up-regulating the expression of c-FLIP.
FAU - Hou, Yusen
AU  - Hou Y
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Chai, Jiake
AU  - Chai J
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China. Email: cjk304@126.com.
FAU - Liu, Lingying
AU  - Liu L
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Yu, Yonghui
AU  - Yu Y
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Duan, Hongjie
AU  - Duan H
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Hu, Quan
AU  - Hu Q
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Chu, Wanli
AU  - Chu W
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
FAU - Ma, Li
AU  - Ma L
AD  - Department of Burn & Plastic Surgery, First Affiliated Hospital, PLA General 
      Hospital, Beijing 100048, China.
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhonghua Yi Xue Za Zhi
JT  - Zhonghua yi xue za zhi
JID - 7511141
RN  - 0 (Endotoxins)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
SB  - IM
MH  - Apoptosis/*drug effects
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Endotoxins/*adverse effects
MH  - Humans
MH  - Lipopolysaccharides/*pharmacology
MH  - Mesenchymal Stem Cells/cytology/*drug effects
MH  - NF-kappa B/metabolism
MH  - Signal Transduction
MH  - Umbilical Cord/cytology
EDAT- 2014/05/24 06:00
MHDA- 2015/02/06 06:00
CRDT- 2014/05/24 06:00
PHST- 2014/05/24 06:00 [entrez]
PHST- 2014/05/24 06:00 [pubmed]
PHST- 2015/02/06 06:00 [medline]
PST - ppublish
SO  - Zhonghua Yi Xue Za Zhi. 2014 Apr 1;94(12):948-51.