白细胞介素-10对大鼠肝脏星状细胞生物活性的影响

In current study, we modified the methods of isolating and identifying rat HSC. The yield rate and cell viability of HSC was improved. Furthermore, a rat HSC cell line which could maintain the characteristics of activation was established through successive passage. The influences of interleukin-10(IL-10) on the biological activity of isolated rat HSC were estimated using cell and molecular biological experiments. The results were as follow: IL-10 could inhibit the expression of intercellular adhesion molecule-1(ICAM-1) in HSC. It also could inhibit the activation of HSC induced by Kupffer cells(KC) through inhibiting of the cytokine secretion of KC. Moreover, under the successful construction of adeno-associated virus（AAV） vector and completion of its packaging and purification, we transduced AAV-mediated IL-10 gene to isolated rat HSC and observed its expression. After being transduced with IL-10, the expression of collagen type I and type III in HSC was decreased while the expression of inducible nitric oxide synthase (iNOS) was increased. We set up the silicone-rubber -membrane method, with which we found that histamine significantly stimulates HSC contraction in a dose dependent way. The effect was blocked by histamine 1-receptor(H1R) antagonist but not histamine 2-receptor antagonist. .Conclusion: 1. The modified isolating method of rat HSC was convenient, efficient and reliable. 2. HSC cell line we established can be served as a model of rat activated HSC and used for in vitro studies of liver fibrogenesis. 3. IL-10 can affect the biological activity of HSC through a direct or indirect pathway, which might play a protective role against liver inflammation and fibrosis. 4. IL-10 can effectively transduced IL-10 gene into HSC, so it can be used as a useful tool for in vivo and in vitro studies of IL-10. 5. Histamine can induce contraction of HSC via H1R, which might play a role in the pathogenesis of portal hypertension. 6. H1R antagonist could block the contraction effect of histamine on HSC, it might become a new agent in therapeutic of portal hypertension..

本研究用含人生野生型IL-10 cDNA腺相关病毒载体转染体外培养的正常细胞及肝硬化大鼠肝星状细胞(HSC)，观察内皮素-1及NO分泌活性，细胞收缩能力及增殖变化。并用α-SMA肌动鞍谆蜃銎舳犹匾煨砸糏L-10基因在肝硬化大鼠肝脏HSC中的表达，探讨HSC在门静脉高压症发病中的地位以及通过调控HSC生物学行为的途径预防和逆转门静脉高压症的可靠性。.