[Interleukin-10 inhibits the activation of cultured rat hepatic stellate cells induced by Kupffer cells].
OBJECTIVE
To investigate the effect of interleukin-10 (IL-10) on the activation of cultured rat hepatic stellate cells (HSC) induced by Kupffer cells (KC) and relevant mechanisms.


METHODS
HSC and KC were isolated and purified from rat liver by collagenase IV perfusion and density gradient centrifugation with Nycodenz. After primary culture for 2 days, HSC and KC were divided randomly into three groups: HSC group, KC group and HSC + KC group (coculture group), then were stimulated by IL-10 of concentrations of 2 ng/ml or 20 ng/ml respectively. After 2 days, the proliferation of HSC were determined with (3)H-TdR incorporating test, the expression of alpha-smooth muscle actin (alpha-SMA) in HSC was detected by Western blotting, and tumor necrosis factor-alpha (TNF-alpha) protein concentration in the supernatant was determined by ELISA.


RESULTS
IL-10 showed no significant effect on HSC proliferation (P > 0.05) and alpha-SMA expression (P > 0.05) in HSC group. The levels of proliferation and alpha-SMA expression of HSC in coculture group were 2.4 times (P < 0.001) and 6 times (P < 0.001) higher respectively than that in HSC groups. In coculture group, HSC proliferation and alpha-SMA expression were decreased by 23% (P < 0.001) and 35% (P < 0.05) respectively after stimulation of 2 ng/ml IL-10, and were decreased by 33% (P < 0.001) and 49% (P < 0.05) respectively after stimulation of 20 ng/ml IL-10 in a dose dependent way. TNF-alpha was not detected in HSC group. The level of TNF-alpha in HSC + KC coculture group was 74% higher than that in KC group (P < 0.01). 2 ng/ml and 20 ng/ml IL-10 reduced the TNF-alpha level by 27% (P < 0.01) and 36% (P < 0.01) respectively in coculture group, and reduced the TNF-alpha level by 29% (P < 0.05) and 42% (P < 0.01) respectively in KC group in a dose dependent way.


CONCLUSIONS
IL-10 reduces the level of TNF-alpha secreted by KC dose-dependently. Through reducing cytokine production by KC, IL-10 inhibits the activation of cultured rat HSC induced by KC, which may play a protective role against liver fibrosis.
Weng
Shangeng
S
Department of Surgery, People's Hospital, Peking University, Beijing 100044, China.
Leng
Xisheng
X
Wei
Yuhua
Y
D000818:Animals; D002478:Cells, Cultured; D016753:Interleukin-10 / Q000494:pharmacology*; D007728:Kupffer Cells / Q000187:drug effects* / Q000502:physiology; D008099:Liver / Q000166:cytology*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology
D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10