Role Of Nuclear Factor-κB In Passway Of Survival/Apoptosis Of HUVEC Regulated By Angiotensin Ⅱ

[Objective] To investigate the role of nuclear factor-kB in the passway of Survival/Apoptosis of HUVEC regulated by AngiotensinⅡ. [Methods] The cell lines of HUVEC cultured in vitro were divided into three groups, respectively pretreated with AngⅡ, AngⅡ+Gliotoxin, and normal culture medium. We mainly investigated the effects of AngⅡ(0．01μmol/L,0．1μmol/L,1μmol/L and 10μmol/L) on the viability of HUVEC at 6^th, 12^th, 18^th, 24^th hour respectively with modified MTT and microscope. Then agarose gel electrophoresis was applied for the detection of DNA ladder in apoptotic HUVEC. Flow cytometry was also used to quantitatively analyze the apoptosis of HUVEC which had been pretreated with 10μmol/L AngⅡfor 24 hours. Finally, the nuclear translocation of NF-kB subunit p65 was evaluated by using immunocytochemistry techniques. The statistical analysis was carried out by using SPSS 11．5 for Windows. Data were statistically analyzed by using one-way ANOVA, followed by Post Hoc Multiple Comparison Test. P<0．05 was considered statistically significant. [Results]①HUVEC were incubated with 1μmol/LAngⅡfor 24 hours, the cell viability decreased significantly (P<0．05 vs normal control group). 10μmol/L AngⅡfor 18 hours and 24 hours, and the cell viability rate decreased significantly (P<0．01 vs normal control group). The deceasing effect of 10μmol/L AngⅡon HUVEC reached maximum at 24^th hour, while 0．1mg/L Gliotoxin antagonized this effect.②DNA ladder appeared in the agarose gel when HUVEC were pretreated with 10μmol/L AngⅡfor 24 hours; while there was not obvious DNA ladder in normal control group and Gliotoxin group.③The results of flow cytometry quantitatively showed that more cells were induced apoptosis with 10μmol/L AngⅡ, and the rate of apoptosis was significantly higher than normal control group (P<0．05 vs normal control group). Meanwhile, there was no significant difference between nonmal control group and Gliotoxin group (P>0．05) .④The analysis of immunocytochemistry techniques suggested that compared with normal control group, NF-kB in HUVEC were increasingly activated by 10μmol/L AngⅡ, and the difference was considered significant (P<0．05) . In contrast, Gliotoxin inhibited the activation of NF-kB. [Conclusions]①AngⅡmay induce changes of morphocytology, decrease cell viability, and lead to apoptosis of HUVEC, and the effect is related with the enlargement of dose of AngⅡ.②Specific inhibitor Gliotoxin of NF-kB antagonizes the effect of AngⅡon HUVEC.③Nuclear Factor-kB may play an important role in the passway of survival/apoptosis of HUVEC regulated by AngiotensinⅡ.