Cellular localization and functional significance of CYP3A4 in the human epileptic brain.
Cellular localization and functional significance of CYP3A4 in the human epileptic brain.
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CYP3A4 在人类癫痫脑中的细胞定位和功能意义。
DOI：
10.1111/j.1528-1167.2010.02956.x
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发表时间：
2011-03
期刊：
影响因子：
5.6
通讯作者：
Janigro D
中科院分区：
文献类型：
--
作者：
Ghosh C;Marchi N;Desai NK;Puvenna V;Hossain M;Gonzalez-Martinez J;Alexopoulos AV;Janigro D
令人信服的证据支持中枢神经系统(CNS)中存在P450酶(Cyps)。然而，关于Cyps在耐药癫痫脑中的定位和功能的信息很少。我们已经评估了特异性酶CYP3A4的表达模式，并研究了其与MDR1的共存。我们还确定了细胞色素P3A4的表达和细胞存活之间是否存在关联。脑组织标本取自8名接受手术以缓解抗药性癫痫或切除海绵状血管瘤的患者。每个样本被分割用于免疫染色或原代培养人内皮细胞和星形胶质细胞。用抗CYP3A4、mdr1、GFAP或NeuN抗体进行免疫染色。采用高效液相色谱-紫外法测定卡马西平(CBZ)在细胞内的代谢。细胞色素P3A4的表达与细胞活力的标志物DAPI缩合相关。将细胞色素P3A4基因导入人胚胎肾(HEK)细胞，进一步研究细胞色素P3A4水平、CBZ代谢和细胞活力之间的关系。血脑屏障(BBB)内皮细胞和大多数神经元(75±10%)表达CYP3A4。免疫荧光染色显示细胞色素P3A4和多药耐药基因mdr1在血管内皮细胞和神经元中共同表达。CYP3A4的表达与DAPI核固缩呈负相关。细胞色素P3A4在HEK细胞中的过表达使其对卡马西平的细胞毒性水平产生抵抗。细胞色素P3A4水平与卡马西平代谢量呈正相关。细胞色素P3A4在脑内的表达不仅与药物代谢有关，而且可能代表一种细胞保护机制。细胞色素P3A4和多药耐药基因mdr1的共同表达可能参与了病变脑组织细胞的存活。
Compelling evidence supports the presence of P450 enzymes (CYPs) in the central nervous system (CNS). However, little information is available on the localization and function of CYPs in the drug-resistant epileptic brain. We have evaluated the pattern of expression of the specific enzyme CYP3A4 and studied its co-localization with MDR1. We also determined whether an association exists between CYP3A4 expression and cell survival. Brain specimens were obtained from eight patients undergoing resection to relieve drug-resistant seizures or to remove a cavernous angioma. Each specimen was partitioned for either immunostaining or primary culture of human endothelial cells and astrocytes. Immunostaining was performed using anti-CYP3A4, MDR1, GFAP, or NeuN antibodies. High performance liquid chromatography–ultraviolet (HPLC-UV) analysis was used to quantify carbamazepine (CBZ) metabolism by these cells. CYP3A4 expression was correlated to DAPI condensation, a marker of cell viability. Human embryonic kidney (HEK) cells were transfected with CYP3A4 to further evaluate the link between CYP3A4 levels, CBZ metabolism, and cell viability. CYP3A4 was expressed by blood–brain barrier (BBB) endothelial cells and by the majority of neurons (75 ± 10%). Fluorescent immunostaining showed coexpression of CYP3A4 and MDR1 in endothelial cells and neurons. CYP3A4 expression inversely correlated with DAPI nuclear condensation. CYP3A4 overexpression in HEK cells conferred resistance to cytotoxic levels of carbamazepine. CYP3A4 levels positively correlated with the amount of CBZ metabolized. CYP3A4 brain expression is not only associated with drug metabolism but may also represent a cytoprotective mechanism. Coexpression of CYP3A4 and MDR1 may be involved in cell survival in the diseased brain.
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影响因子：
5.6
作者：
Ghosh C;Gonzalez-Martinez J;Hossain M;Cucullo L;Fazio V;Janigro D;Marchi N
通讯作者：
Marchi N
影响因子：
7.2
作者：
Koryllou, Angeliki;Patrinou-Georgoula, Meropi;Pletsa, Vassiliki
通讯作者：
Pletsa, Vassiliki
影响因子：
5.6
作者：
LEVY, RH
通讯作者：
LEVY, RH
影响因子：
4.3
作者：
Ma X;Idle JR;Gonzalez FJ
通讯作者：
Gonzalez FJ
影响因子：
4.7
作者：
Dauchy, Sandrine;Dutheil, Fabien;Decleves, Xavier
通讯作者：
Decleves, Xavier