JNK inhibitor AS601245 decreased levels of Bim and CC3 proteins and reduced neuronal apoptosis and brain infarct volume in P7 rat after HI.

<p>Samples were obtained from the cortex. Bim and CC3 protein levels were significantly reduced at 48 h in the AS601245-treated cortex compared with the DMSO-treated cortex (A). Cell apoptosis was detected by TUNEL staining with the In Situ Cell Death Detection Kit (Merck Millipore) according to the manufacturer's protocol. Ten fields were chosen randomly at 400x magnification to count the apoptotic and total cells. The apoptotic index (AI) was calculated as follows: AI = (number of apoptotic cells/total number counted) ×100%. TUNEL-positive cells were not detected in the sham controls (C). However, the positive cells were increased at 6 h (data not shown) and 24 h (data not shown) and peaked at 48 h (D) after HI. Meanwhile, AS601245 pretreatment obviously reduced cellular apoptosis at 48 h after HI (F) <u>a</u>nd decreased the infarct volume 7 days after HI significantly (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132998#pone.0132998.g004" target="_blank">Fig 4H</a>), compared with the DMSO controls (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132998#pone.0132998.g004" target="_blank">Fig 4E and 4H</a>,). The cell apoptosis index in the AS601245-treated brain cortex was decreased by 59% compared with the DMSO-treated brain cortex (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132998#pone.0132998.g004" target="_blank">Fig 4G</a>). The infarct volume in the AS601245-treated brain was decreased by 58% compared with the DMSO controls (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0132998#pone.0132998.g004" target="_blank">Fig 4I</a>).