Race and sex differences in ROS production and SOD activity in HUVECs

Abstract

Black individuals and men are predisposed to an earlier onset and higher prevalence of hypertension, compared with White individuals and women, respectively. Therefore, the influence of race and sex on reactive oxygen species (ROS) production and superoxide dismutase (SOD) activity following induced inflammation was evaluated in female and male human umbilical vein endothelial cells (HUVECs) from Black and White individuals. It was hypothesized that HUVECs from Black individuals and male HUVECs would exhibit greater ROS production and impaired SOD activity. Inflammation was induced in HUVEC cell lines (n = 4/group) using tumor necrosis factor-alpha (TNF-α, 50ng/ml). There were no between group differences in ROS production or SOD activity in HUVECs from Black and White individuals, and HUVECs from Black individuals exhibited similar SOD activity at 24hr compared with 4hr of TNF-α treatment (p>0.05). However, HUVECs from White individuals exhibited significantly greater SOD Activity (p<0.05) at 24hr as compared to 4hr in the control condition but not with TNF-α treatment (p>0.05). Female HUVECs exhibited significantly lower ROS production than male HUVECs in the control condition and following TNF-α induced inflammation (p<0.05). Only female HUVECs exhibited significant increases in SOD activity with increased exposure time to TNF-α induced inflammation (p<0.05). HUVECs from White individuals alone exhibit blunted SOD activity when comparing control and TNF-α conditions. Further, compared to female HUVECs, male HUVECs exhibit a pro-inflammatory state.

Fig 1

CellROX/ Hoechst (ROS production normalized to living cells) in HUVECs from Black (B) and White (W) individuals at 4 hours and 24 hours of time-matched control (panel A) and TNF-α treatment (panel B). n = 11 for Control 4hr B HUVECs and TNF-α 4hr B HUVECs, n = 12 for all other groups and times. * denotes p<0.05 from post-hoc t-tests and significant model effects are denoted by † (time), # (sex), or ^ (interaction).

Fig 2

SOD activity normalized to protein content (U/mg) in HUVECs from Black (B) and White (W) individuals at 4hr and 24hr for control (panel A) and TNF-α treatment (panel B). n = 9 for all groups and times. * denotes p<0.05 from post-hoc t-tests and significant model effects are denoted by † (time), # (sex), or ^ (interaction).

Fig 3

CellROX/ Hoechst (ROS production normalized to living cells) in female (F) and male (M) HUVECs at 4 hours and 24 hours of time-matched control (panel A) and TNF-α treatment (panel B). n = 11 for Control 4hr F HUVECs and TNF-α 4hr F HUVECs, n = 12 for all other groups and times. * denotes p<0.05 from post-hoc t-tests and significant model effects are denoted by † (time), # (sex), or ^ (interaction).

Fig 4

SOD activity normalized to protein content (U/mg) in female (F) and male (M) HUVECs at 4 hours and 24 hours of time-matched control (panel A) and TNF-α treatment (panel B). n = 9 for all groups and times. * denotes p<0.05 from post-hoc t-tests and significant model effects are denoted by † (time), # (sex), or ^ (interaction).