Effect of coenzyme I on apoptosis of liver cell line L02 induced by ischemia-reperfusion injury.
OBJECTIVE
To study the protective effect of coenzyme I (NADH) on normal liver cell line L02 against ischemia-reperfusion injury.


METHODS
Cultured L02 cells was divided into 3 groups, namely ischemia-reperfusion group (I/R), ischemia-reperfusion group with NADH pretreatment (NADH+I/R) and control group. Assisted by flow cytometry (FCM), the apoptosis rate and the expression of apoptosis-related proteins (Bcl-2, p16, p21, p53) were detected in L02 cells in the 3 groups. Apoptotic L02 cells were also observed under transmission electron microscope.


RESULTS
I/R significantly inhibited L02 cell proliferation and increased their apoptosis rate up to (31.53+/-8.27)% 12 h after I/R. In the presence of NADH, however, the apoptotic rate of the cells was significantly decreased to (12.61+/-2.34)% (P<0.05). FCM indicated that I/R down-regulated the expression of Bcl-2 and up-regulated the expression of p16, p21 and p53 proteins as compared with control group, but NADH acted to the reverse effect. Typical apoptotic features were observed by transmission electron microscope in the cells 12 h after I/R.


CONCLUSIONS
I/R might induce apoptosis in L02 cells by up-regulating the expression of p16, p21, p53 and down-regulating the expression of Bcl-2, and the protective effect of NADH against I/R-induced apoptosis may lie in the expression regulation of the proteins concerned.
Xu
Xiao-Ping
XP
Department of Hepatobiliary Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. xuchopin@163.net
Li
Kai-Zong
KZ
Dou
Ke-Feng
KF
Liu
Fa-Quan
FQ
Zhang
Ji-Ren
JR
D017209:Apoptosis; D002460:Cell Line; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000096:biosynthesis; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008854:Microscopy, Electron; D009243:NAD / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D015427:Reperfusion Injury; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
C493956:CDKN1A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D009243:NAD