摘要目的：检测自噬相关蛋白Beclin-1、LC3及p62在难治性癫痫相关皮质发育畸形病变中的表达情况，初步探讨皮质发育畸形出现细胞水平异常的可能机制。方法选用18例手术切除的致痫灶标本，包括结节硬化综合征（ TSC）、局灶性皮质发育不良（ FCD）Ⅱb型及FCDⅠ型各6例。采用免疫组织化学染色（ EnVison二步法）对3种蛋白分别在3组病变中细胞水平的表达情况进行定性分析，并计数Beclin-1、LC3染色阳性细胞数进行比较；应用Western blot方法对LC3蛋白在3组病变中的表达进行定量比较分析。结果免疫组织化学染色结果显示3种蛋白均主要表达于FCDⅡb型及TSC组中的形态异常神经元和气球样细胞／巨大细胞内。 Beclin-1为胞质内颗粒状或弥漫阳性，另可见到轴索强阳性表达；LC3为胞质内弥漫阳性或核周阳性；p62为胞质内弥漫阳性，部分为核及核周阳性。此外，形态异常神经元中Beclin-1、LC3及p62阳性程度高于气球样细胞／巨大细胞。而作为对照的FCDⅠ组中仅有个别或部分细胞呈弱阳性表达。 Beclin-1和LC3染色中阳性细胞数均为TSC组＞FCDⅡb组＞FCDⅠ组，且TSC组、FCDⅡb组与FCDⅠ组的差异均具有统计学意义（ P＜0.05）。Western blot结果显示LC3蛋白在病灶组织内的表达量为 TSC组＜FCDⅡb组＜FCDⅠ组。结论TSC、FCDⅡb中的形态异常神经元和气球样细胞／巨大细胞中存在自噬抑制，推测自噬的异常可能与蛋白质的异常堆积密切相关；但形态异常神经元和气球样细胞中自噬异常具体机制不完全一致。

abstractsObjective To study the expression of autophagy-related proteins ( Beclin-1, LC3 and p62) in brain tissue with malformations of cortical development and related molecular pathogenesis. Methods The brain tissue of 18 cases with epileptogenic foci resection, including 6 cases of tuberous sclerosis complex (TSC), 6 cases of focal cortical dysplasia type Ⅱb (FCD Ⅱb) and 6 cases of focal cortical dysplasia type Ⅰ ( FCD Ⅰ) , were retrieved.Immunohistochemical study for Beclin-1, LC3 and p62 proteins was performed.The degree of positivity for Beclin-1 and LC3 proteins was compared.Western blot was used to quantitatively analyze the LC3 protein in focal lesion of each disease groups.Results Immunohistochemical study showed that the three proteins were mainly expressed in the dysmorphic neurons and balloon cells/giant cells of TSC and FCD Ⅱb.The positivity was more intense in the dysmorphic neurons than the other cell types.Immunostaining for Beclin-1 showed granular or diffuse cytoplasmic positivity, in addition to the strong expression in axons.On the other hand, LC3 showed diffuse or perinuclear cytoplasmic expression.The staining for p62 was mainly cytoplasmic or perinuclear and sometimes nuclear.In FCD typeⅠ, only individual cells showed positive expression for the three proteins.The number of Beclin-1 and LC3-positive cells was larger in TSC group, followed by FCDⅡb group and FCDⅠgroup.And there were significant differences between TSC group and FCDⅠgroup, as well as FCDⅡb group and FCDⅠgroup (P<0.05).Quantitative expression of LC3 protein by Western blot showed smaller amount in TSC group, followed by FCD Ⅱb group and FCDⅠ group.Conclusions The dysmorphic neurons and balloon cells/giant cells of TSC and FCD Ⅱb show abnormality in autophagy, resulting in intracytoplasmic protein accumulation.There are differences in molecular pathogenesis in these cell types.