Ting Zhou1, Xue Zhou1, Bin Song1. 1. Dept. of Stomatology, Guizhou Provincial People's Hospital, Guiyang 550002, China.
Abstract
OBJECTIVES: To investigate the role and molecular mechanism of necrostatin-1 (Nec-1), a specific programmed cell necrosis inhibitor, in promoting the oxidative stress response of macrophages under high glucose (HG) environment. METHODS: Macrophages were cultured in control (5.5 mmol·L-1 glucose) or HG (25 mmol·L-1 glucose) medium for 72 h. The HG+Nec-1 group was given HG and 5 μmol·L-1 Nec-1. Reactive oxygen species (ROS) level, malondialdehyde (MDA) activity, and superoxide dismutase (SOD) activity were measured by 2'-7'dichlorofluorescin diacetate, MDA, and SOD enzyme linked immunosorbent assay kits, respectively. Moreover, receptor interacting protein 1 (RIP1) expression was assessed through real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot (WB). Finally, after the expression of RIP1 in macrophages was silenced, the effect of HG environment on oxidative stress response was evaluated in the gene-deficient cells. RESULTS: The HG group had increased ROS level and MDA activity (P<0.000 1) and decreased SOD activity (P<0.000 1) compared with the control group. The HG+Nec-1 group had higher ROS level and MDA activity (P<0.000 1) and lower SOD activity (P<0.01) than the HG group. The qRT-PCR and WB results showed that RIP1 mRNA level (P<0.001) and protein expression level (P<0.000 1) in the HG group were significantly higher than those in the control group, and RIP1 mRNA and protein expression levels in the HG+Nec-1 group were significantly lower than those in the HG group (P<0.000 1). After RIP1 was silenced effectively (P<0.001) with si-RNA, the ROS level and MDA activity of the HG+si-RIP1 group decreased compared with those of the HG+si-negative control (si-NC) group (P<0.001), and SOD activity in the HG+si-RIP1 group increased than that in the HG+si-NC group (P<0.000 1). CONCLUSIONS: HG promotes oxidative stress on macrophages by upregulating RIP1 expression.
OBJECTIVES: To investigate the role and molecular mechanism of necrostatin-1 (Nec-1), a specific programmed cell necrosis inhibitor, in promoting the oxidative stress response of macrophages under high glucose (HG) environment. METHODS: Macrophages were cultured in control (5.5 mmol·L-1 glucose) or HG (25 mmol·L-1 glucose) medium for 72 h. The HG+Nec-1 group was given HG and 5 μmol·L-1 Nec-1. Reactive oxygen species (ROS) level, malondialdehyde (MDA) activity, and superoxide dismutase (SOD) activity were measured by 2'-7'dichlorofluorescin diacetate, MDA, and SOD enzyme linked immunosorbent assay kits, respectively. Moreover, receptor interacting protein 1 (RIP1) expression was assessed through real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot (WB). Finally, after the expression of RIP1 in macrophages was silenced, the effect of HG environment on oxidative stress response was evaluated in the gene-deficient cells. RESULTS: The HG group had increased ROS level and MDA activity (P<0.000 1) and decreased SOD activity (P<0.000 1) compared with the control group. The HG+Nec-1 group had higher ROS level and MDA activity (P<0.000 1) and lower SOD activity (P<0.01) than the HG group. The qRT-PCR and WB results showed that RIP1 mRNA level (P<0.001) and protein expression level (P<0.000 1) in the HG group were significantly higher than those in the control group, and RIP1 mRNA and protein expression levels in the HG+Nec-1 group were significantly lower than those in the HG group (P<0.000 1). After RIP1 was silenced effectively (P<0.001) with si-RNA, the ROS level and MDA activity of the HG+si-RIP1 group decreased compared with those of the HG+si-negative control (si-NC) group (P<0.001), and SOD activity in the HG+si-RIP1 group increased than that in the HG+si-NC group (P<0.000 1). CONCLUSIONS: HG promotes oxidative stress on macrophages by upregulating RIP1 expression.
Entities:
Keywords:
diabetes; necrostatin-1; oxidative stress; periodontitis; receptor interacting protein 1
Authors: E Bernabe; W Marcenes; C R Hernandez; J Bailey; L G Abreu; V Alipour; S Amini; J Arabloo; Z Arefi; A Arora; M A Ayanore; T W Bärnighausen; A Bijani; D Y Cho; D T Chu; C S Crowe; G T Demoz; D G Demsie; Z S Dibaji Forooshani; M Du; M El Tantawi; F Fischer; M O Folayan; N D Futran; Y C D Geramo; A Haj-Mirzaian; N Hariyani; A Hasanzadeh; S Hassanipour; S I Hay; M K Hole; S Hostiuc; M D Ilic; S L James; R Kalhor; L Kemmer; M Keramati; Y S Khader; S Kisa; A Kisa; A Koyanagi; R Lalloo; Q Le Nguyen; S D London; N D Manohar; B B Massenburg; M R Mathur; H G Meles; T Mestrovic; A Mohammadian-Hafshejani; R Mohammadpourhodki; A H Mokdad; S D Morrison; J Nazari; T H Nguyen; C T Nguyen; M R Nixon; T O Olagunju; K Pakshir; M Pathak; N Rabiee; A Rafiei; K Ramezanzadeh; M J Rios-Blancas; E M Roro; S Sabour; A M Samy; M Sawhney; F Schwendicke; F Shaahmadi; M A Shaikh; C Stein; M R Tovani-Palone; B X Tran; B Unnikrishnan; G T Vu; A Vukovic; T S S Warouw; Z Zaidi; Z J Zhang; N J Kassebaum Journal: J Dent Res Date: 2020-03-02 Impact factor: 6.116