摘要
AIM: To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells.METHODS: Rat hepatic stellate cells (HSCs) were isolated and purified from rat liver by in situ digestion of collagenase and pronase and single-step density Nycodenz gradient.After activated by culture in vitro, HSCs were divided into 4 groups and treated with nothing (group N), PDGF (group P),IL-10 (group I) and PDGF in combinalJon with IL-10 (group C),respectively. Semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was employed to compare the mRNA expression levels of Fas/FasL and Bcl-2/Bax in HSCs of each group.RESULTS: The expression levels of Fas between the 4 groups had no significant differences (P>0.05). FasL mRNA level in normal culture-activated HSCs (group N) was very low.It increased obviously after HSCs were treated with IL-10(group I) (0.091±0.007 vs 0.385±0.051, P<0.01), but remained the low level after treated with PDGF alone (group P)or PDGF in combination with IL-10 (group C). Contrast to the control group, after treated with PDGF and IL-10, either alone or in combination, Bcl-2 mRNA expression was downregulated and Bax mRNA expression was up-regulated, both following the turn from group P, group I to group C.Expression of Bcl-2 mRNA in group C was significantly lower than that in group P (0.126±0.008 vs0.210±0.024, P<0.01).But no significant difference was found between group C and group I, as well as between group I and group P (P>0.05).Similarly, the expression of Bax in group C was higher than that in group P (0.513±0.016 vs0.400±0.022, P<0.01).No significant difference was found between group I and group P (P>0.05). But compared with group C, Bax expressions in group I tended to decrease (0.449±0.028 vs 0.513±0.016,P<0.05).CONCLUSION: PDGF may promote proliferation of HSCs but is neutral with respect to HSC apoptosis. IL-10 may promote the apoptosis of HSCs by up-regulating the expressions of FasL and Bax and down-regulating the expression of Bcl-2, which may be involved in its antifibrosis mechanism.
AIM:To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells. METHODS:Rat hepatic stellate cells (HSCs) were isolated and purified from rat liver by in situ digestion of collagenase and pronase and single-step density Nycodenz gradient. After activated by culture in vitro,HSCs were divided into 4 groups and treated with nothing (group N),PDGF (group P), IL-10 (group I) and PDGF in cornbination with IL-10 (group C), respectively.Semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was employed to compare the mRNA expression levels of Fas/FasL and Bcl- 2/Bax in HSCs of each group. RESULTS:The expression levels of Fas between the 4 groups had no significant differences (P>0.05).FasL mRNA level in normal culture-activated HSCs (group N) was very low. It increased obviously after HSCs were treated with IL-10 (group I) (0.091±0.007 vs 0.385±0.051,P<0.01),but remained the low level after treated with PDGF alone (group P) or PDGF in combination with IL-10 (group C).Contrast to the control group,after treated with PDGF and IL-10,either alone or in combination,Bcl-2 mRNA expression was down- regulated and Bax mRNA expression was up-regulated,both following the turn from group P,group I to group C. Expression of Bcl-2 mRNA in group C was significantly lower than that in group P (0.126±0.008 vs 0.210±0.024,P<0.01). But no significant difference was found between group C and group I,as well as between group I and group P (P>0.05). Similarly,the expression of Bax in group C was higher than that in group P (0.513±0.016 vs 0.400±0.022,P<0.01). No significant difference was found between group I and group P (P>0.05).But compared with group C,Bax expressions in group I tended to decrease (0.449±0.028 vs 0.513±0.016, P<0.05). CONCLUSION:PDGF may promote proliferation of HSCs but is neutral with respect to HSC apoptosis.IL-10 may promote the apoptosis of HSCs by up-regulating the expressions of FasL and Bax and down-regulating the expression of Bcl-2,which may be involved in its antifibrosis mechanism.
基金
Supported by the Science and Technology Fundation of FujianProvince,No.2003D05
