Cell lines

Human neuroblastoma cells (SH-SY5Y )

Preparation Method

SH-SY5Y cells were treated with 100 μM glutamate in the presence or absence of Mito-Tempo for 24 h. The cell viability was assessed by the mitochondrial performance, using the MTT reduction assay.

Reaction Conditions

50, 100 μM Mito-Tempo for 24h.

Applications

10 and 100 μM glutamate significantly reduced the number of the living cell to 88.86 ± 3.45% and 51.12 ± 2.97%. 50 and 100 μM Mito-Tempo significantly restored cell viability to 82.90 ± 1.78% and 93.56 ± 2.85%, respectively, indicating that Mito-Tempo could protect the cells from glutamate toxicity.

Animal models

C57BL/6 mice

Preparation Method

Lipopolysaccharide- (LPS-) induced experimental sepsis mouse model was established by injecting male mice intraperitoneally with LPS. Mito-TEMPO was intraperitoneally injected 1 h prior to LPS injection.

Dosage form

20 mg/kg Mito-TEMPO, intraperitoneal(i.p.) injection

Applications

In the LPS+Mito-TEMPO group, the levels of alanine transaminase(ALT) and aspartate transaminase (AST) were approximately 1.5- and 2-fold lower compared with those in the LPS group. Mito-TEMPO pretreatment protects mice against LPS-Induced acute liver injury.

• Iscience 25.1 (2022): 103558.

Mito-Tempo is a mitochondria-targeted antioxidant with effective superoxide scavenging properties, which converts toxic superoxide molecules into hydrogen peroxide or oxygen and subsequently detoxified to oxygen and water by catalase or glutathione peroxidase^[1].

In vitro, Mito-Tempo has a greater protective effect by enhancing superoxide dismutase (SOD) activity and PI3K/AKT/mTOR phosphorylation. Glutamate-exposed cells significantly increased cellular oxidative stress by enhancing ROS production. Glutamate treatment also increased LDH release follows the loss of mitochondrial membrane potential, caused cell viability loss. Treatment with Mito-Tempo not only attenuated the generation of ROS and improved mitochondrial membrane potential but also reduced the neurotoxicity of glutamate in a concentration-dependent manner, which leads to increased cell viability and decreased LDH release^[2]

In vivo，Mito-TEMPO pretreatment inhibited inflammation, attenuated LPS-induced liver injury, and enhanced the antioxidative capability in septic mice, as evidenced by the decreased MDA content and the increased SOD activity. In addition, Mito-TEMPO restored mitochondrial size and improved mitochondrial function. Finally, we found that the levels of pyroptosis-related proteins in the liver of LPS-treated mice were lower after pretreatment with Mito-TEMPO^[3]

References:
[1]. Liang HL, Sedlic F, et al. SOD1 and MitoTEMPO partially prevent mitochondrial permeability transition pore opening, necrosis, and mitochondrial apoptosis after ATP depletion recovery. Free Radic Biol Med. 2010 Nov 30;49(10):1550-60.
[2]. Mukem S, Thongbuakaew T, et al. Mito-Tempo suppresses autophagic flux via the PI3K/Akt/mTOR signaling pathway in neuroblastoma SH-SY5Y cells. Heliyon. 2021 Jun 15;7(6):e07310.
[3]. Mukem S, Thongbuakaew T, et al. Mito-Tempo suppresses autophagic flux via the PI3K/Akt/mTOR signaling pathway in neuroblastoma SH-SY5Y cells. Heliyon. 2021 Jun 15;7(6):e07310.