Effects of HIV-1 Tat protein on cell viability and oxidative stress of U87 cells
10.3760/cma.j.cn112866-20200829-00232
- VernacularTitle:HIV-1 Tat蛋白对U87细胞活性及氧化应激的影响
- Author:
Shuaizhi GUO
1
;
Dandi LI
;
Zeming QIN
;
Hongling WEN
;
Zhiyu WANG
;
Tao HUANG
;
Li ZHAO
Author Information
1. 山东大学公共卫生学院微生物检验学系 山东省"十三五"高校重点实验室，济南 250012
- Keywords:
HIV-associated dementia;
HIV-1 Tat protein;
Malondialdehyde;
Glutathione peroxidase
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(1):34-38
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the amino acid site variation of HIV-1 Tat protein from different parts of AIDS patients with HAD and non HAD and its effect on oxidative stress of U87 cells.Methods:HIV-1 Tat amino acid sequences were analyzed by BLAST and MEGA6 software to study the variation of amino acid sites in four parts of central nervous tissue basal ganglia(BG) and peripheral spleen (SPL) of an HIV-associated dementia (HAD) patient(H) and a non-HAD patient(N), The HIV-1 tat genes were transfected into U87 cell. The green fluorescent protein was observed under microscope to determine the Tat protein expression. The expression of Tat protein in U87 cells was detected by Western blotting. CK-8 method , Western blotting and malondialdehyde (MDA) detection kit were used to study the effect of Tat protein on cell activity, oxidative stress index glutathione peroxidase (GPX), MDA level. Results:Amino acid sequence analysis showed that the key amino acid sites of HIV-1 Tat protein from N-BG, N-SPL, H-BG and H-SPL were different; Tat protein could inhibit the activity of U87 cells, which could be reversed by antioxidant N-acetyl-L-cysteine (NAC). Compared with the control group, the levels of MDA were increased and the expression of GPX protein was decreased in the four experimental groups ( P<0.05). And different sources of Tat protein had different ability to induce oxidative stress, the level of MDA in H-BG group was higher than that in N-BG group( P<0.05). The expression of GPX protein in BG group of both HAD and non-HAD patients was lower than that of SPL group( P<0.05). Conclusions:There are differences in the key amino acid sites of Tat protein in peripheral and central nervous system between HAD and non-HAD patients, and their effects on oxidative stress were also different.
