化学缺氧对小鼠胰腺星状细胞增殖活化的影响

摘要

目的 探讨化学缺氧对小鼠胰腺星状细胞(pancreatic stellate cells,PSCs)增殖活化的影响.方法 原代培养出小鼠PSCs并进行鉴定,于培养基中加入CoCl2(使其在培养基中的终浓度分别为0、50、100、200 μmol/L)培养18 h以诱导化学低氧状态,免疫组化检测低氧诱导因子-1α(HIF-1α)表达情况,免疫荧光测定PSCs活化情况,细胞计数明确细胞增殖情况.结果 随着培养基中CoCl2终浓度的增加,表达HIF-1α阳性的细胞百分比逐渐增加,由0增加至32.000％(31.325％,34.175％)(H=42.333,P=0.000).活化的PSCs比例逐渐增加,由(3.833±2.234)％上调至(21.950±1.446)％(F=258.774,P=0.000),PSCs计数逐渐增加,由(61.08±2.712)×104 ml-1增加至(71.67±2.309)×104 ml-1(F=28.967,P=0.000).结论 缺氧条件下通过HIF-1α引起PSCs增殖及活化程度的增加.%Objective To investigate the effects of hypoxia on the proliferation and activation of pancreatic stellate cells(PSCs).Methods The mouse PSCs were isalated and indentified,when reached confluence,the cells were incubated in the DMEM with cobalt chloride(to make the final concentration to 0,50,100,200 μmol/L)for 18 hours.The expression of HIF-1α was investigated by immunohistochemistry.Then the α-SMA expression was determined by immunofluorescence staining.And the cell proliferation was defined by the method of cell count.Results When the final concentration of CoCl2 solution were increased from 0 to 200 μmol/L,the expression of HIF-1α was increased from 0 to 32.000％(31.325％,34.175％).The difference among these four groups was significant(H=42.333,P=0.000);after 18 hours,the expressions of α-SMA were increased:(3.833±2.234)％for 0 μmol/L and(21.950±1.446)％for 200 μmol/L(F=258.774,P=0.000).The number of PSCs also showed an increasing trend,the final concentration of CoCl2 from 0 to 200 μmol/L,cell counts were increased from(61.08±2.712)×104 ml-1 to(71.67±2.309)×104 ml-1(F=28.967,P=0.000).Conclusion The proliferation and activation of PSCs increase significantly under hypoxia condition;HIF-1α is involved in the pathophysiological process of proliferation and activation of PSCs.