Effect of tumor necrosis factor-α and interleukin-1β on bovine pu lmonary arterial endothelial cell apoptosis

Objective]To investigate the effect of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) on apoptosis of bovine pulmonary arterial endothelial cells(BPEC). MethodsBPECs were incubated with TNF-α,IL-1β or the combination of TNF-α and IL-1β. BEPCs were also pretreated with TNF-α monoclonal antibody or Ac-DEVD-CHO(caspase-3 inhibitor),and followed by incubation with TNF-α. The apopt otic rate was measured by flow cytometry(Annexin V-FITC and PI staining). Results(1)TNF-α induced apoptosis in a dose-dependent manner after a 24 h incubation. With the increase of the TNF-α(500,1 000,2 000 U/ml respectively),the BPEC apoptotic rateTo investigate the effect of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) on apoptosis of bovine pulmonary arterial endothelial cells(BPEC). MethodsBPECs were incubated with TNF-α,IL-1β or the combination of TNF-α and IL-1β. BEPCs were also pretreated with TNF-α monoclonal antibody or Ac-DEVD-CHO(caspase-3 inhibitor),and followed by incubation with TNF-α. The apopt otic rate was measured by flow cytometry(Annexin V-FITC and PI staining). Results(1)TNF-α induced apoptosis in a dose-dependent manner after a 24 h incubation. With the increase of the TNF-α(500,1 000,2 000 U/ml respectively),the BPEC apoptotic rate[(8.21±0.70)%,(9.63±0.71)%,(17.43±1.99)%,respectively] was significantly higher than that of the control group[(3.09±0.08)%, P0.05].(2)TNF-α induced apoptosis was time dependent. After incubated with TNF-α 2 000 U/ml,the BPEC apoptotic rate[(6.72±0.38)%,(7.72±1.66)%,(12.95±0.32)%,(17.70±1.79)%, P[WTBZ]0.05] increased significantly with time increased significantly with time[6,12,24,36 h,respectively] of TNF-α incubation.(3)The BPEC apoptotic rate of anti-TNF-α monoclonal antibody group[(7.78±0.21)%] or the Ac-DEVD-CHO group[(7.32±0.11)%] was significantly higher than the TNF-α group alone[(10.59±0.49)%, PThere is nosynergistic effect betweenI L 1βand TNF αin the induction of BPE Capoptosis