Hijmans Jamie G, Stockleman Kelly, Reiakvam Whitney, Levy Ma'ayan V, Brewster Lillian M, Bammert Tyler D, Greiner Jared J, Connick Elizabeth, DeSouza Christopher A
Department of Integrative Physiology, Integrative Vascular Biology Laboratory, University of Colorado, Boulder, Colorado.
Department of Medicine, Division of Infectious Disease, University of Arizona, Tucson, Arizona.
Physiol Rep. 2018 Mar;6(6):e13647. doi: 10.14814/phy2.13647.
The aim of this study was to determine, in vitro, the effects of X4 and R5 HIV-1 gp120 and Tat on: (1) endothelial cell senescence and (2) endothelial cell microRNA (miR) expression. Endothelial cells were treated with media without and with: R5 gp120 (100 ng/mL), X4 gp120 (100 ng/mL), or Tat (500 ng/mL) for 24 h and stained for senescence-associated β-galactosidase (SA-β-gal). Cell expression of miR-34a, miR-217, and miR-146a was determined by RT-PCR. X4 and R5 gp120 and Tat significantly increased (~100%) cellular senescence versus control. X4 gp120 significantly increased cell expression of miR-34a (1.60 ± 0.04 fold) and miR-217 (1.52 ± 0.18), but not miR-146a (1.25 ± 0.32). R5 gp120 significantly increased miR-34a (1.23 ± 0.07) and decreased miR-146a (0.56 ± 0.07). Tat significantly increased miR-34a (1.49 ± 0.16) and decreased miR-146a (0.55 ± 0.23). R5 and Tat had no effect on miR-217 (1.05 ± 0.13 and 1.06 ± 0.24; respectively). HIV-1 gp120 (X4 and R5) and Tat promote endothelial cell senescence and dysregulation of senescence-associated miRs.
本研究的目的是在体外确定X4和R5 HIV-1 gp120以及Tat对以下方面的影响：（1）内皮细胞衰老；（2）内皮细胞微小RNA（miR）表达。用不含以及含有R5 gp120（100 ng/mL）、X4 gp120（100 ng/mL）或Tat（500 ng/mL）的培养基处理内皮细胞24小时，并对衰老相关β-半乳糖苷酶（SA-β-gal）进行染色。通过逆转录聚合酶链反应（RT-PCR）测定miR-34a、miR-217和miR-146a的细胞表达。与对照组相比，X4和R5 gp120以及Tat显著增加（约100%）细胞衰老。X4 gp120显著增加miR-34a（1.60±0.04倍）和miR-217（1.52±0.18）的细胞表达，但未增加miR-146a（1.25±0.32）的表达。R5 gp120显著增加miR-34a（1.23±0.07）并降低miR-146a（0.56±0.07）。Tat显著增加miR-34a（1.49±0.16）并降低miR-146a（0.55±0.23）。R5和Tat对miR-217无影响（分别为1.05±0.13和1.06±0.24）。HIV-1 gp120（X4和R5）以及Tat可促进内皮细胞衰老和衰老相关miR的失调。
