西红花酸对乙醛诱导的肝星状细胞增殖和胶原合成的影响

目的: 探讨西红花酸(Crocetin)对乙醛刺激的大鼠肝星状细胞(Hepatic stellate cell, HSC)增殖和胶原合成的影响及其作用机制。方法: 培养大鼠肝星状细胞HSC-T6,建立乙醛诱导的HSC纤维化模型;用不同浓度的西红花酸(10^-6、10^-7、10^-8 mol/L)对乙醛刺激的HSC-T6进行处理,MTT法检测细胞增殖;羟脯氨酸测定检测HSC-T6胶原含量;流式细胞分析仪测定细胞凋亡;Western blot检测细胞ERK1/2、Bax、Bcl-2蛋白的表达;RT-PCR检测Ⅰ型、Ⅲ型胶原蛋白、间质胶原酶(MMP-2)、组织金属蛋白酶抑制因子-1(TIMP-1)的基因表达。结果: 在一定浓度范围里,西红花酸能抑制乙醛引起的HSC增殖和胶原合成; 西红花酸诱导乙醛刺激的HSC细胞凋亡;西红花酸能增加Bax蛋白表达,降低乙醛刺激升高的ERK1/2、Bcl-2蛋白表达;西红花酸能明显降低Ⅰ型、Ⅲ型胶原、TIMP-1的表达,提高MMP-2的表达。结论: 西红花酸通过抑制乙醛诱导的HSC增殖和胶原合成以及促进活化的HSC凋亡起到抗肝纤维化作用,其机制可能与ERK信号传导通路和对基质金属蛋白酶的调节有关。

Effects of crocetin on proliferation and collagen synthesis of hepatic stellate cell

AIM: To investigate the effects of crocetin on proliferation, collagen synthesis in hepatic stellate cell (HSC) stimulated by acetaldehyde and its partial mechanism.METHODS: HSC stimulated by acetaldehyde was incubated with different doses of crocetin(10^-8,10^-7,10^-6 mol/L), cell proliferation was analyzed by MTT assay, apoptosis ratio was analyzed by flow cytometry(FCM), synthesis of collagen was observed by hydroxyproline concentration measurement. The ERK1/2, Bax, Bcl-2 were tested by Western blotting. Matrix metalloproteinase(MMP-2),the tissue inhibitor metalloproteinase (TIMP-1) mRNA, collagen mRNA were examined by RT-PCR.RESULTS: Within a concentration coverage, crocetin inhibited proliferation of HSC induced by acetaldehyde. The apoptosis rate of cells in crocetin treated groups increased with concentration compared with acetaldehyde group. Crocetin decreased the levels of ERK1/2 and Bcl-2 expression, but increased Bax protein expression. Crocetin could markedly suppressed the up-regulation of CollagenⅠmRNA, Collagen ⅢmRNA, TIMP-1 mRNA, while increased MMP-2 mRNA expression.CONCLUSION: Crocetin inhibited HSC proliferation and collagen synthesis stimulated by acetaldehyde, which related to the expression of ERK1/2 and the regulation of extracellular matrix.