Protective Effect of Tempol on Buthionine Sulfoximine-Induced Mitochondrial Impairment in Hippocampal Derived HT22 Cells

Abstract EN

Using a simulated oxidative stress model of hippocampus-derived immortalized cell line (HT22), we report that prooxidant buthionine sulfoximine (BSO, 1 mM, 14 h), without adversely affecting cell viability or morphology, induced oxidative stress by inhibiting glutathione synthesis.

BSO treatment also significantly reduced superoxide dismutase (SOD) activity ( p < 0.05 ) and significantly lowered total antioxidant capacity ( p < 0.001 ) in HT22 cells when compared to vehicle treated control cells.

Antioxidant tempol, a piperidine nitroxide considered a SOD mimetic, reversed BSO-induced decline in SOD activity ( p < 0.01 ) and also increased BSO-induced decline in total antioxidant capacity ( p < 0.05 ).

Interestingly, BSO treatment significantly reduced mitochondrial oxygen consumption ( p < 0.05 ), decreased mitochondrial membrane potential ( p < 0.05 ), and lowered ATP production ( p < 0.05 ) when compared to vehicle treated control cells, collectively indicative of mitochondrial impairment.

Antioxidant tempol treatment mitigated all three indicators of mitochondrial impairment.

We postulate that BSO-induced oxidative stress in HT22 cells caused mitochondrial impairment, and tempol by increasing SOD activity and improving antioxidant capacity presumably protected the cells from BSO-induced mitochondrial impairment.

In conclusion, present study provides an interesting simulation of oxidative stress in hippocampal cells, which will serve as an excellent model to study mitochondrial functions.