血小板衍生生长因子和白细胞介素-10对体外培养的大鼠肝星状细胞中Fas/Fas配体及Bcl-2/Bax mRNA表达的影响
Effects of platelet-derived growth factor and interleukin-10 on Fas/Fas-ligand and Bcl-2/Bax mRNA expression in rat hepatic stellate cells in vitro.
作者信息
Wang Xiao-Zhong, Zhang Sheng-Jun, Chen Yun-Xin, Chen Zhi-Xin, Huang Yue-Hong, Zhang Li-Juan
机构信息
Department of Gastroenterology, Union Hospital of Fujian Medical University, Fuzhou 350001, Fujian Province, China.
出版信息
World J Gastroenterol. 2004 Sep 15;10(18):2706-10. doi: 10.3748/wjg.v10.i18.2706.
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 combination 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.
目的
探讨血小板衍生生长因子（PDGF）和白细胞介素-10（IL-10）对大鼠肝星状细胞中Fas/Fas配体及Bcl-2/Bax mRNA表达的影响。
方法
采用胶原酶和链霉蛋白酶原位消化及一步密度Nycodenz梯度法从大鼠肝脏分离纯化肝星状细胞（HSCs）。体外培养激活后，将HSCs分为4组，分别给予无处理（N组）、PDGF（P组）、IL-10（I组）及PDGF与IL-10联合处理（C组）。采用半定量逆转录聚合酶链反应（RT-PCR）分析比较各组HSCs中Fas/FasL及Bcl-2/Bax的mRNA表达水平。
结果
4组间Fas表达水平无显著差异（P>0.05）。正常培养激活的HSCs（N组）中FasL mRNA水平很低。HSCs经IL-10处理后（I组）明显升高（0.091±0.007对0.385±0.051，P<0.01），但单独用PDGF处理（P组）或PDGF与IL-10联合处理（C组）后仍维持低水平。与对照组相比，单独或联合应用PDGF和IL-10处理后，Bcl-2 mRNA表达下调，Bax mRNA表达上调，均呈现从P组、I组到C组的变化趋势。C组Bcl-2 mRNA表达显著低于P组（0.126±0.008对0.210±0.024，P<0.01）。但C组与I组、I组与P组之间差异无统计学意义（P>0.05）。同样，C组Bax表达高于P组（0.513±0.016对0.400±0.022，P<0.01）。I组与P组之间差异无统计学意义（P>0.05）。但与C组相比，I组Bax表达有降低趋势（0.449±0.028对0.513±0.016，P<0.05）。
结论
PDGF可能促进HSCs增殖，但对HSC凋亡呈中性作用。IL-10可能通过上调FasL和Bax表达、下调Bcl-2表达促进HSCs凋亡，这可能参与其抗纤维化机制。
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