Caspase抑制剂Ⅰ对体外培养肝细胞的抗凋亡作用

The anti-apoptotic effects of Caspase inhibitor Ⅰ on cultured hepatocytes

目的探讨Caspase抑制剂Ⅰ(ZVADfmk)对肝细胞的抗凋亡作用。方法用不同浓度(0～1000ngml)的重组可溶性FasL或穿孔素(20ngml)加不同浓度(0～10μgml)的重组粒酶B诱导体外培养肝细胞的凋亡。检测肝细胞凋亡的百分率及肝细胞凋亡的效应酶Caspase3的活性。结果实验组肝细胞的凋亡百分率及Caspase3的活性稳定在低水平(P=007,P=092);而对照组肝细胞的凋亡百分率和Caspase3的活性明显升高(P<001),而且随着重组可溶性FasL和重组粒酶B浓度的升高而上升。在相同浓度的重组可溶性FasL或重组粒酶B诱导凋亡下,实验组的肝细胞发生凋亡的百分率和Caspase3活性明显低于对照组(P<001,P<0001)。结论Caspase抑制剂Ⅰ(ZVADfmk)对体外培养的肝细胞具有明显的抗凋亡作用。

Objective To investigate the anti apoptotic effects of Caspase inhibitor Ⅰ (ZVAD fmk) on cultured hepatocytes Methods Apoptosis of cultured hepatocytes was induced with different concentration of recombinant soluble FasL (0～1?000 ng/ml) or perforin (20 ng/ml) plus different concentration of recombinant granzyme B (0～1 0 μg/ml) Caspase inhibitor I was added to incubate with hepatocytes for 24 hrs or 1 hr in the experimental group The percentage of hepatocyte apoptosis and the activity of Caspase 3 were detected Results Hepatocytes in the control group showed a dose dependent increase of percentage in TUNEL positive staining and Caspase 3 activity ( P=0.07, P= 0 92), while those in the experimental group remained at a low level of TUNEL positive staining and Caspase 3 activity ( P >0 05) Under the same treatment concentration of rsFasL or granzyme B, percentage in TUNEL positive staining and Caspase 3 activity in the experimental group were much lower than those in the control group ( P <0 01, P <0 001) Conclusion Caspase inhibitor I (ZVAD fmk) can prevent the in vivo cultured hepatocytes from apoptosis induced by rsFasL or granzyme B