International Journal of Pharma and Bio Sciences
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10.22376/ijpbs.2019.10.1.p1-12
Volume 7 Issue 4
2016 (October - December)
Tertiary-butyl hydroperoxide (tbh), mediated oxidative stress may lead to alteration in necrotic and apoptotic gene expression in n2a cell lines.
Tertiary-butyl hydroperoxide (TBH) is a chemical that can cause oxidative stress in vitro through catalysis by glutathione peroxidase to form GSSG and tertiary butyl alcohol. N2a cell lines are a fast-growing mouse neuroblastoma cell line and have been commonly used to study neurotoxicity. We aimed for present study to clarify whether; Tertiary-butyl hydroperoxide (TBH), mediated oxidative stress may lead to alteration in necrotic and apoptotic gene expression in N2a cell lines. The study involve following two groups: Group- 1: N2a without TBH exposure; Group -2: N2a with 45 min 62.5 micro molar concentration TBH exposure and reperfusion for 48 hrs. RT PCR analysis was done for the following gene; Degree of oxidative stress (TRX1&2, PRX); Type of cell death (PARP 1; Caspase 3), Cell defense (HSP 70, HO 1); Housekeeping (18 S RNA). We observed, serum deprivation for 45 min affects N2A cells to some extend as evident from "rounding-off" seen in them, while phase image did not show much difference between these and TBH treated cell. RT-PCR analysis showed increased in PARP-1, and decrease in HSP-70, Trx-2, and Prxgene expression which signifies cell death through necrosis. After 3 days of re-perfusion with normal growth medium (with serum), cells treated with TBH showed extensive loss. This could be due to the washing off of dead cells during medium changes (Cell death might be due to initial necrosis and also due to delayed apoptosis). Delayed apoptosis was evident from increased Caspase 3 expression from RT-PCR. We concluded that there was a gene expression change during the acute and sub-acute phase of the injury which mimics the in-vivo scenario of reperfusion after tissue insult and during the acute phase of stress the cell death occurs through necrosis and then gets shifted to apoptosis during sub-acute phase.
GOWTHAM PADMANABAN AND DRAVIDA MANI ELUMALAI
Apoptosis; Necrosis; N2a cell; Oxidative stress, TBH.
152-157