Role and mechanism of necrostin-1 in promoting oxidative stress response of macrophages in high glucose condition

doi:10.7518/hxkq.2021.06.008

Abstract

Abstract: Objective

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).

Conclusion

HG promotes oxidative stress on macrophages by upregulating RIP1 expression.

Key words: necrostatin-1, diabetes, periodontitis, oxidative stress, receptor interacting protein 1

CLC Number:

R 781.4

Zhou Ting, Zhou Xue, Song Bin. Role and mechanism of necrostin-1 in promoting oxidative stress response of macrophages in high glucose condition[J]. West China Journal of Stomatology, 2021, 39(6): 675-681.

Figures/Tables 6

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