摘要
目的 :研究乙醛对新分离的肝星状细胞 (hepaticstellatecell,HSC)增殖、表达α1(Ⅰ )型胶原mRNA、IkB和NF -kB的影响 ,以探讨酒精性肝纤维化发病机理。方法 :用链霉蛋白酶、胶原酶以及密度梯度离心法分离培养大鼠肝星状细胞。采用四甲基偶氮唑蓝 (MTT)比色法、原位杂交法、免疫细胞化学法、凝胶电泳迁移率法 (EMSA)分别检测肝星状细胞的增殖、胶原合成、IkBα以及NF -kB的表达。结果 :(1)成功分离出大鼠肝星状细胞。 (2 )乙醛刺激新分离培养的肝星状细胞后细胞明显增殖 ;乙醛刺激传一代肝星状细胞后 ,α1(Ⅰ )型胶原mRNA表达显著增加 ;IkBα表达明显下降 ;乙醛作用肝星状细胞后 3 0min ,NF -kB的表达即有增加 ,到 12 0min时达最高值。结论 :乙醛可以使静止的肝星状细胞活化 ,表现为增殖、α1(Ⅰ )型胶原mRNA合成、IkBα表达下降、NF -kB活性增加。
Objective:To study the effects of acetaldehyde on the proliferation, synthesis of collagen gene mRNA, expression of IkB and NF-kB in ratshepatic stellate cells(HSC). Method:HSC were isolated by density gradient centrifugation after digested by pronase E and collagenase IV, and then the relationships between acetaldehyde and prolifeation, synthesis of collagen gene, expression of IkB and NF-kB were investigated by means of MTT, immunocytochemistry , in situ hybridization and EMSA. Result: The freshly isolated HSC proliferated significantly after 24 h treated with 200μM acetaldehyde. The expression of α1(I) collagen mRNA increased aparently after 12h treated with acetaldehyde. After 12h treated with acetaldehyde, the expression of IkB decreased, and the activity of NF-kB reached to the peak. Conclusion: Acetaldehyde could activate quiescent HSC, and induced proliferation of HSC, synthesis of collagen gene and expression of NF-kB, and reduced the activity of IkB.
出处
《西南国防医药》
CAS
2002年第2期97-100,共4页
Medical Journal of National Defending Forces in Southwest China
基金
全军"十五"医药卫生科研基金资助 NO :0 1MB0 37
