【摘要】 肝病是威胁人类健康的主要疾病之一,而肝具有强大的再生能力,因此开发肝再生的药物靶标对肝病防治具有重大意义。本室前期采用大鼠全基因组芯片检测发现,丝氨酸蛋白酶抑制剂Kazal型Ⅲ(serine protease inhibitor Kazal typeⅢ,SPINK3)在大鼠肝再生中表达显著改变。研究表明,SPINK3是一类结构与表皮生长因子(epidermal growth factor,EGF)相似的生长因子,能够与表皮生长因子受体(EGFR)结合促进细胞增殖。本文通过基因过表达和干涉的方法处理原代大鼠肝细胞,通过CCK8法、Ki67免疫荧光法、PI单染法和Annexin V/PI双染法检测SPINK3表达变化对原代大鼠肝细胞活力、增殖、周期和凋亡的影响。结果显示SPINK3过表达时能够显著提高原代大鼠肝细胞的细胞活力,促进其细胞周期和增殖,并抑制其细胞凋亡,而干涉SPINK3表达则显著降低原代大鼠肝细胞的细胞活力,抑制其细胞周期和增殖,并促进其细胞凋亡。以上结果表明,SPINK3能够促进体外培养的原代大鼠肝细胞的增殖,并抑制其凋亡。更多还原

【Abstract】 Liver disease is one of the major diseases threatening human health nowadays.The liver has a strong ability to regenerate.Therefore,the development of drug target in liver regeneration plays an important role on liver disease prevention and therapy.Previous studies found that serine protease inhibitor Kazal type Ⅲ(SPINK3) was significantly changed in rat liver regeneration by analysis of gene expression profiles.SPINK3 is not only a trypsin inhibitor,but also a growth factor similar to epidermal growth factor(EGF).SPINK3 can bind to epidermal growth factor receptor(EGFR) and promote cell proliferation.In this study,interference of SPINK3 expression was achieved by transfection of adenovirus vectors with either insertion or deletion of SPINK3 into rat primary hepatocytes.Cell viability was detected by CCK8 and cell proliferation was detected by Ki67 immunofluorescence.In addition,cell cycle was detected by PI single staining and cell apoptosis was detected by Annexin V/PI double staining.The results showed that SPINK3 overexpression increased the cell viability of primary rathepatocytes,promoted cell cycle and inhibited apoptosis of the cells,while knock down of SPINK3 expression reduced the cell viability,inhibited cell cycle and promoted apoptosis of the cells.更多还原