ABSTRACT
Objective To study the effects of hypoxia on the cell migration,apoptosis and expression of related genes and proteins of human dermal microvascular endothelial cells(HDMEC). Methods To culture HDMEC in hypoxia condition by putting CoCl2in the cell incubator and all the subjects were divided to two groups which included normal group and hypoxia group.To find the appropriate CoCl2dose by CCK-8 experiment and the 200 μmol/L CoCl2was the best to establish hypoxia model.The migration of cells was measured by wound healing test.Apoptosis rate was detec-ted by flow cytometry. The gene expressions were detected by RT-PCR and the protein levels were detected by West-ern blot. Results The migration ability of cells was enhanced in hypoxia condition (P<0.05). The apoptosis rate was increased in hypoxia condition(P<0.05).The gene expressions of HIF-1α,HIF-1β,VEGF,iNOS were increased comparing with normal groups depending on time(P<0.05).The protein expressions of HIF-1α,VEGF,iNOS were in-creased comparing with normal groups depending on time(P<0.05). Conclusions Cell migration, apoptosis of HDMEC were influenced by hypoxia. It may be explained by increasing expression of related gene and proteins like HIF-1α,VEGF,iNOS in hypoxia condition.
ABSTRACT
Objective To study the effects of hypoxia on the cell migration,apoptosis and expression of related genes and proteins of human dermal microvascular endothelial cells(HDMEC). Methods To culture HDMEC in hypoxia condition by putting CoCl2in the cell incubator and all the subjects were divided to two groups which included normal group and hypoxia group.To find the appropriate CoCl2dose by CCK-8 experiment and the 200 μmol/L CoCl2was the best to establish hypoxia model.The migration of cells was measured by wound healing test.Apoptosis rate was detec-ted by flow cytometry. The gene expressions were detected by RT-PCR and the protein levels were detected by West-ern blot. Results The migration ability of cells was enhanced in hypoxia condition (P<0.05). The apoptosis rate was increased in hypoxia condition(P<0.05).The gene expressions of HIF-1α,HIF-1β,VEGF,iNOS were increased comparing with normal groups depending on time(P<0.05).The protein expressions of HIF-1α,VEGF,iNOS were in-creased comparing with normal groups depending on time(P<0.05). Conclusions Cell migration, apoptosis of HDMEC were influenced by hypoxia. It may be explained by increasing expression of related gene and proteins like HIF-1α,VEGF,iNOS in hypoxia condition.