[The effect of IL-10 on proliferation of primary cultured hepatocytes].
OBJECTIVE
To investigate the effect of IL-10 on proliferation of rat primary cultured hepatocytes.


METHODS
Rat hepatocytes were isolated from rat liver by in situ digestion of collagenase IV and cryopreserved, resuscitated, cultured in vitro. Reverse-transcription polymerase chain reaction (RT-PCR) was used to characterize the purity of hepatocytes and analyze IL-10/IL10Ralpha mRNA from freshly isolated cells. The primary cultured hepatocytes were divided into 3 groups and treated with nothing (group N), Insulin (group C), and IL-10 in combination with Insulin (group I), respectively. Nuclear cell cycle analysis, MTT, and Trypan Blue cell count was assayed.


RESULTS
RT-PCR showed expression of characterization genes in primary hepatocytes group and liver tissue are different. RT-PCR showed an expression of IL-10/IL10Ralpha mRNA in rat primary hepatocytes. Trypan Blue cell count showed an depression of cell quantity in group I at 48h (71.96% contrast to group C, P<0.05). MTT analyse also showed absorbance of group I was declined contrast to group C at 24 h and 48 h (88.41% and 90.24%, P<0.05). Cell cycle analysis via FCM showed a decline at 24h in group I than group C and group N (59.06% and 70.18%, P<0.01).


CONCLUSIONS
The primary hepatocytes we isolated is quite purity. Rat primary hepatocytes express IL-10/IL10Ralpha mRNA. IL-10 has an suppression effect on proliferation of primary cultured hepatocytes.
Lin
Xian-Ping
XP
Department of Gastroenterology, Union Hospital, Fujian Medical University, Fuzhou 350001, China. linxiangping001@163.com
Huang
Yue-Hong
YH
Zheng
Wei-da
WD
Chen
Yun-Xin
YX
Chen
Zhi-Xin
ZX
Wang
Xiao-Zhong
XZ
D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D007328:Insulin / Q000494:pharmacology; D016753:Interleukin-10 / Q000235:genetics / Q000494:pharmacology*; D053709:Interleukin-10 Receptor alpha Subunit / Q000235:genetics; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
D007328:Insulin; D053709:Interleukin-10 Receptor alpha Subunit; D016753:Interleukin-10