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凋亡在癫痫发生机制中起重要作用,但其在难治性癫痫耐药机制中的作用尚不清楚.为研究X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)反义寡核苷酸对K562/Dox(阿霉素诱导)耐药细胞及难治性癫痫大鼠耐药性的影响,首先建立耐药的K562/Dox细胞株,比较XIAP在耐药细胞株和正常K562细胞株的表达情况,观察转染XIAP反义寡核苷酸后,线粒体膜电位变化以及对卡马西平和苯妥英钠耐药性的影响.另外,建立慢性杏仁核点燃癫痫模型,筛选出耐药组和药物敏感组,通过侧脑室注射XIAP反义寡核苷酸,对照组注射生理盐水.观察其对各组大鼠后放电阈值(after dischargethreshold,ADT)、后放电时程(after discharge duration,ADD)等电生理指标的影响.结果发现,XIAP在K562/Dox耐药细胞上的表达明显高于正常K562细胞,XIAP反义寡核苷酸转染K562/Dox耐药细胞后,XIAP的表达明显下降.导致了K562/Dox细胞线粒体跨膜电位的下降,而且对苯妥英钠和卡马西平的耐药性明显下降,IC50分别由(1978.2±90.3)mg/L和(1875.6±83.2)mg/L,降低到(1123.5±54.2)mg/L和(1084.5±60.6)mg/L,逆转倍数分别为1.76和1.73.同时动物实验发现,耐药组大鼠在给予XIAP反义寡核苷酸后,ADT明显高于对照组(P<0.05),ADD时程也明显缩短.上述结果证明,XIAP在耐药的K562/Dox细胞株存在高表达,下调XIAP在K562/Dox细胞株表达可以改善K562/Dox对卡马西平和苯妥英钠的耐药性.而且下调XIAP表达可以协助AEDs改善耐药大鼠的电生理活动,提示XIAP参与了难治性癫痫的耐药.
Apoptosis plays an important role in the pathogenesis of epilepsy, but its role in the mechanism of refractory epilepsy is unclear.In order to study the antisense oligodeoxynucleotide (X-linked inhibitor of apoptosis protein, XIAP) (K562 / Dox) induced drug-resistant cells and refractory epilepsy rats, we first established the drug-resistant K562 / Dox cell lines and compared XIAP expression in drug-resistant cell lines with normal K562 Cell lines and observe the changes of mitochondrial membrane potential and the resistance to carbamazepine and phenytoin after the transfection of XIAP antisense oligonucleotides.Moreover, we established a model of chronic amygdaloid-ignited epilepsy and screened drug-resistant Group and drug-sensitive group, injected XIAP antisense oligonucleotide into the lateral ventricle, and the control group was injected with normal saline. The effects of different doses of XIAP on post-discharge duration (ADT), post-discharge duration ADD) and other electrophysiological indicators.It was found that the expression of XIAP on K562 / Dox drug-resistant cells was significantly higher than that of normal K562 cells, and XIAP expression was inhibited after XIAP antisense oligonucleotides were transfected into K562 / Dox drug-resistant cells Significant decrease led to mitochondria in K562 / Dox cells Transmembrane potential decreased, and the resistance to phenytoin sodium and carbamazepine was significantly decreased, the IC50 decreased from (1978.2 ± 90.3) mg / L and (1875.6 ± 83.2) mg / L to (1123.5 ± 54.2) mg / L and (1084.5 ± 60.6) mg / L respectively, and the reversal multiples were 1.76 and 1.73, respectively.At the same time, the ADT in the drug-resistant group was significantly higher than that in the control group (P < 0.05), and the duration of ADD significantly shortened.The above results demonstrate that XIAP is highly expressed in the resistant K562 / Dox cell line, down-regulating the expression of XIAP in K562 / Dox cells can improve the inhibitory effect of K562 / Dox on carbamazepine and phenytoin sodium Drug resistance.And down-regulated XIAP expression can help AEDs to improve the electrophysiological activity of drug-resistant rats, suggesting that XIAP is involved in the resistance of refractory epilepsy.