RESUMEN
OBJECTIVE: To study for the expression of heat shock protein (hsp) 90 and hsp27 when human umbilical vein endothelial cell (HUVEC) was treat with heat shock and H2O2. METHODS: We collected HUVEC from fresh umbilical vein within 30 minutes after delivery by 0.25% trypsin and cultured them at 37 degrees C under 5% CO2, using EBM2 media. The cells were fed twice a week with a complete change of fresh culture medium. The cultured cells were identified as endothelial cells by western blot using CD34, and vWF markers. We divided HUVEC as control group applying heat stress at 37 degrees C for 30 minutes, stress group applying heat stress at 43 degrees C for 30 minutes, H2O2 treatment group exposing of 100 micrometer H2O2 for 60 minutes, and heat stress + H2O2 treatment group applying same state of heat stress and exposing of H2O2 for 60 minutes. We extract protein and examine the expression of hsp90 and hsp27 by western blot. RESULTS: HUVEC reached full confluence state in our culture condition at 7 days on 25 mL flask. We observed the constant expression character of CD 34 and vWF in cultured cell. Under the every experimental condition, we observed the expression of hsp90 and hsp27 in HUVEC. And we observed that hsp27 was more strongly expressed in heat shock + H2O2 treatment group and heat shock group than control group and H2O2 treatment group. CONCLUSION: By identifying the expression difference of hsp in vitro oxidative stress, fetal tissue might play the protective role in the intrauterine stress.
Asunto(s)
Humanos , Western Blotting , Células Cultivadas , Células Endoteliales , Feto , Proteínas de Choque Térmico , Calor , Estrés Oxidativo , Choque , Tripsina , Venas UmbilicalesRESUMEN
OBJECTIVE: To study for the expression of heat shock protein (hsp) 90 and hsp27 when human umbilical vein endothelial cell (HUVEC) was treat with heat shock and H2O2. METHODS: We collected HUVEC from fresh umbilical vein within 30 minutes after delivery by 0.25% trypsin and cultured them at 37 degrees C under 5% CO2, using EBM2 media. The cells were fed twice a week with a complete change of fresh culture medium. The cultured cells were identified as endothelial cells by western blot using CD34, and vWF markers. We divided HUVEC as control group applying heat stress at 37 degrees C for 30 minutes, stress group applying heat stress at 43 degrees C for 30 minutes, H2O2 treatment group exposing of 100 micrometer H2O2 for 60 minutes, and heat stress + H2O2 treatment group applying same state of heat stress and exposing of H2O2 for 60 minutes. We extract protein and examine the expression of hsp90 and hsp27 by western blot. RESULTS: HUVEC reached full confluence state in our culture condition at 7 days on 25 mL flask. We observed the constant expression character of CD 34 and vWF in cultured cell. Under the every experimental condition, we observed the expression of hsp90 and hsp27 in HUVEC. And we observed that hsp27 was more strongly expressed in heat shock + H2O2 treatment group and heat shock group than control group and H2O2 treatment group. CONCLUSION: By identifying the expression difference of hsp in vitro oxidative stress, fetal tissue might play the protective role in the intrauterine stress.