CAJ | 학술논문

目的通过对难治性癫痫患者脑组织中P-糖蛋白、肺耐药相关蛋白表达与卡马西平浓度的比较及相关性研究,探讨难治性癫痫耐药的机制.方法选取2007年至2009年间于宣武医院功能神经外科行癫痫致痫灶切除术治疗且术前服用卡马西平半年以上的26例药物难治性癫痫患者的脑组织标本(其中局灶性皮质发育不良ⅠblO例、Ⅱa4例、Ⅱb2例,神经节细胞胶质瘤6例,胚胎发育不良性神经上皮瘤4例).应用Envision二步法进行免疫组织化学标记,观察两种耐药蛋白在脑组织中的表达部位和表达强度.应用Western blot法进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,对两种耐药蛋白在脑组织病灶区域和病灶周围区域的表达分别进行定量分析.应用荧光偏振免疫分析法对病灶区域和病灶周围区域的卡马西平浓度进行测定.结果 P-糖蛋白在局灶性皮质发育不良Ⅱ型和难治性癫痫相关脑肿瘤病例的病灶区域表达(μg/ml)均高于病灶周围区域(分别是2.593±0.829和1.711±0.292,t=-2.201,P=0.028；1.352±0.445和1.179±0.593,t=2.698,P=0.028).肺耐药相关蛋白在局灶性皮质发育不良Ⅱ型和难治性癫痫相关脑肿瘤病例的病灶区域表达(μg/ml)亦均高于病灶周围区域(分别是1.567±0.092和0.775±0.101,t=-2.516,P=0.024;1.091±0.239和0.825±0.297,t =3.997,P=0.003).卡马西平在难治性癫痫相关脑肿瘤病灶区域平均浓度(μg/ml)低于病灶周围区域(0.848±0.726和0.948±0.785,t=-3.056,P=0.014),在病灶区域卡马西平浓度低于病灶周围区域的8例中,病灶区域P-糖蛋白和肺耐药相关蛋白同时升高.但是P-糖蛋白、肺耐药相关蛋白表达量与难治性癫痫患者脑组织卡马西平浓度无显著的等级相关性.结论耐药蛋白参与了难治性癫痫耐药的过程,同时提示癫痫的耐药是一个复杂的过程,不同病变可能有不同的机制参与了耐药过程.
목적 통과대난치성전간환자뇌조직중P-당단백、폐내약상관단백표체여잡마서평농도적비교급상관성연구,탐토난치성전간내약적궤제.방법 선취2007년지2009년간우선무의원공능신경외과행전간치간조절제술치료차술전복용잡마서평반년이상적26례약물난치성전간환자적뇌조직표본(기중국조성피질발육불량ⅠblO례、Ⅱa4례、Ⅱb2례,신경절세포효질류6례,배태발육불량성신경상피류4례).응용Envision이보법진행면역조직화학표기,관찰량충내약단백재뇌조직중적표체부위화표체강도.응용Western blot법진행십이완기류산납-취병희선알응효전영,대량충내약단백재뇌조직병조구역화병조주위구역적표체분별진행정량분석.응용형광편진면역분석법대병조구역화병조주위구역적잡마서평농도진행측정.결과 P-당단백재국조성피질발육불량Ⅱ형화난치성전간상관뇌종류병례적병조구역표체(μg/ml)균고우병조주위구역(분별시2.593±0.829화1.711±0.292,t=-2.201,P=0.028；1.352±0.445화1.179±0.593,t=2.698,P=0.028).폐내약상관단백재국조성피질발육불량Ⅱ형화난치성전간상관뇌종류병례적병조구역표체(μg/ml)역균고우병조주위구역(분별시1.567±0.092화0.775±0.101,t=-2.516,P=0.024;1.091±0.239화0.825±0.297,t =3.997,P=0.003).잡마서평재난치성전간상관뇌종류병조구역평균농도(μg/ml)저우병조주위구역(0.848±0.726화0.948±0.785,t=-3.056,P=0.014),재병조구역잡마서평농도저우병조주위구역적8례중,병조구역P-당단백화폐내약상관단백동시승고.단시P-당단백、폐내약상관단백표체량여난치성전간환자뇌조직잡마서평농도무현저적등급상관성.결론 내약단백삼여료난치성전간내약적과정,동시제시전간적내약시일개복잡적과정,불동병변가능유불동적궤제삼여료내약과정.
Objective To compare the expression and distribution of drug resistance proteins Pglycoprotein (P-gp) and lung resistance protein (LRP) in brain tissues of patients with refractory epilepsy and to investigate the relationship between expression of drug resistance proteins and concentration of antiepileptic drug carbamazepine (CBZ).Methods We included the brain tissues of 26 cases with refractory epilepsy who had in the experiment.They had following pathologic diagnosis: focal cortical dysplasia (FCD) Ⅰb (n =10),FCD Ⅱ a (n =4),FCD Ⅱb (n =2) and brain tumorsincluding ganglioglioma (n =6) and dysembryoplastic neuroepithelial tumor(n =4).Immunohistochemistry staining using EnVision system was used to reveal the expression location of P-gp and LRP,and Western blot in SDS-polyacrylamide gel was used to quantitatively analyze the expression of P-gp and LRP.Fluorescence polarization immunoassay was used to determine concentration of CBZ.Results Both P-gp and LRP performed(μg/ml) prominent overexpression in brain tissues of patients with refractory epilepsy,especially in the lesions of both FCD type Ⅱ (P-gp: 2.593 ±0.829 vs 1.711 ±0.292,t =-2.201,P=0.028;LRP:1.352 ±0.445 vs 1.179 ±0.593, t =-2.516,P =0.028, respectively)and tumor(P-gp:1.567 ±0.092 vs 0.775 ± 0.101, t =2.698, P =0.024; LRP: 1.091 ± 0.239 vs 0.825 ± 0.297, t =3.997, P =0.003respectively).The concentration of CBZ in lesions of brain tumors were lower than which in surrounding regions(0.848 ±0.726 vs 0.948 ±0.785, t =-3.056,P =0.014), while P-gp and LRP were higher than which in surrounding regions in 80％ cases.There was no relationship between expression of drug resistance proteins and concentration of CBZ.Conclusions In processing of drug resistance, P-gp and LRP play important roles.However, there is no correlation between expression of drug resistance proteins and concentration of CBZ, suggesting epilepsy drug resistance to be a complicated mechanism.