Han Jing, Sun Li, Liang Hao, Cheng Yan
Department of Neurology, Tianjin Medical University General Hospital, Tianjin 300052, China.
Department of Neurology, Tianjin Medical University General Hospital, Tianjin 300052, China. Email:
Zhonghua Yi Xue Za Zhi. 2014 Apr 22;94(15):1179-83.
To explore the effects of 12/15-lipoxygenase antisense oligonucleotide (asON-12/15-LOX) on OGD (oxygen-glucose deprivation)-induced PPARγ (peroxisome proliferator-activated receptor γ) expression and nuclear translocation in primarily cultured cortical neurons.
After a 48-h pre-treatment of 12/15-LOX antisense oligonucleotide (asON), primarily cultured cortical neurons underwent 3-hour OGD followed by a 24-hour reperfusion.Immunofluorescent staining and Western blot were used to evaluate the expressions of 12/15-LOX and PPARγ as well as the nuclear translocation of PPARγ.
Compared with the control group, the expressions of 12/15-LOX and PPARγ whole protein were enhanced in OGD group (t = -3.72 and -6.79, P = 0.03 and 0.04). And an increase of PPARγ in nucleus (t = -4.67, P = 0.02) could be noted with a simultaneous reduction in cytosol (t = 3.40, P = 0.04) after OGD, indicating an induction of nuclear translocation by OGD. Compared with OGD group, a pre-treatment of asON-12/15-LOX dramatically attenuated OGD-induced increase in 12/15-LOX whole protein expression (t = 5.03, P = 0.02). Compared with OGD group, a pre-treatment of asON-12/15-LOX greatly reduced OGD-induced increase in PPARγ total protein expression (t = 2.83, P = 0.04) and nuclear translocation (t = 7.05, P = 0.01 for nuclear protein; t = -5.47, P = 0.01 for cytosol protein). It indicated a possible link between 12/15-LOX and PPARγ.
12/15-LOX antisense oligonucleotide suppresses the expression and nuclear translocation of PPARγ in primarily cultured cortical neurons after OGD.
探讨12/15-脂氧合酶反义寡核苷酸（asON-12/15-LOX）对氧糖剥夺（OGD）诱导的原代培养皮层神经元中过氧化物酶体增殖物激活受体γ（PPARγ）表达及核转位的影响。
对原代培养的皮层神经元进行12/15-LOX反义寡核苷酸（asON）预处理48小时后，进行3小时的氧糖剥夺，随后再灌注24小时。采用免疫荧光染色和蛋白质免疫印迹法评估12/15-LOX和PPARγ的表达以及PPARγ的核转位情况。
与对照组相比，OGD组中12/15-LOX和PPARγ全蛋白表达增强（t = -3.72和-6.79，P = 0.03和0.04）。OGD后可观察到细胞核中PPARγ增加（t = -4.67，P = 0.02），同时细胞质中PPARγ减少（t = 3.40，P = 0.04），表明OGD诱导了核转位。与OGD组相比，asON-12/15-LOX预处理显著减弱了OGD诱导的12/15-LOX全蛋白表达增加（t = 5.03，P = 0.02）。与OGD组相比，asON-12/15-LOX预处理大大降低了OGD诱导的PPARγ总蛋白表达增加（t = 2.83，P = 0.04）以及核转位（核蛋白t = 7.05，P = 0.01；细胞质蛋白t = -5.47，P = 0.01）。这表明12/15-LOX与PPARγ之间可能存在联系。
12/15-LOX反义寡核苷酸可抑制OGD后原代培养皮层神经元中PPARγ的表达及核转位。
