摘要
目的探讨白细胞介素18(IL-18)及IL-18抗体抗免疫性肝纤维化药血清对离体小鼠肝星状细胞(mHSC)增殖、凋亡的影响。方法取12周末正常对照(A)组,兔IgG(B)组,刀豆蛋白A(C)组,IL-18(D)组,IL-18抗体(E)组小鼠注射药物24 h后血清。用MTT法观察药血清对小鼠mHSC增殖作用,荧光显微镜、DNA凝胶电泳、流式细胞仪分析凋亡作用,半定量RT-PCR检测药物血清处理的mHSC 72 h基质金属酶2(MMP-2)mRNA,基质金属酶抑制剂1(TIMP-1)mRNA表达。结果与A、B组相比,C、D、E组药血清可以促进mHSC的增殖(P<0.05),E组较C组抑制mHSC作用明显(P<0.05)。以0.5 pg/ml～50μg/ml浓度的IL-18和IL-18抗体观察对mHSC的增殖作用不明显。荧光显微镜观察到mHSC核的浓染致密、固缩、片段化以及新月型改变。A、B、C、D、E组mHSC的平均凋亡率为(12.55±2.31)%、(13.01±2.32)%(、6.59±1.06)%(、4.26±0.63)%、(9.04±1.62)%,组间具有统计学差异(P<0.05)。C、D、E组MMP-2 mRNA,TIMP-1 mRNA表达较A、B组明显(P<0.05)。结论 IL-18药血清可以促进mHSC的增殖,并增加MMP-2 mRNA和TIMP-1 mRNA的表达。
Objective To explore the effects of the serum with anti-immunohepatofibrotic drugs of interleukin（IL）-18 or IL-18 antibody on the apoptosis and proliferration of mice hepatic stellate cells（mHSC）.Methods The serum was obtained from 12 weeks mice at 24 hours after mice were repeatedly injected normal saline（group A）,rabbit IgG（group B）,concanavalin A（group C）,IL-18（group D） or anti-IL-18（group E）.The proliferration of mHSC was observed using MTT assay and cell apoptosis was analyzed using flow cytometry and fluorescent microscopy.MMP-2 mRNA and TIMP-1 mRNA were measured by quantitative reverse transcripation-PCR.Results The proliferation of mHSC in serum was significantly higher in groups of C,D and E than that in groups of A and B（P〈0.05）,which was lower in group E than that in group C（P〈0.05）.The increase in proliferation of mHSC was not significant in groups D and E.The dense and compact fluorescence particles and alteration nucleus of mHSC were observed under microscope.The apoptosis rates of groups of A,B,C,D and E were（12.55±2.31）%,（13.01±2.32）%,（4.26±0.63）%,（6.59±1.06）% and （9.04±1.62）%（P〈0.05）.The expressions of MMP-2 mRNA and TIMP-1 mRNA were higher in groups of C,D and E than those in groups of A and B（P〈0.05）.Conclusion The serum of IL-18 can promote the proliferation of mHSC and increase the expressions of MMP-2 mRNA and TIMP-1 mRNA.
出处
《江苏医药》
CAS
CSCD
北大核心
2011年第17期1991-1994,I0001,共5页
Jiangsu Medical Journal
基金
镇江市社会发展项目(SH20070074)
