促肾上腺皮质激素释放激素及受体在婴儿痉挛症致痫组织中的表达

Expression of corticotropin releasing hormone and its receptors in infantile spasm

目的婴儿痉挛症(Infantile spasm,IS),又称West综合症,属于年龄依赖性癫痫综合症,脑组织促肾上腺皮质激素释放激素(Corticotropin releasing hormone,CRH)多被认为是IS的可能致病机制,目前尚未在人IS患者致痫灶中得到直接依据。试验研究IS患者手术切除致痫灶中CRH的表达,探讨CRH信号途径与IS癫痫痉挛发作及发病机制的相关性。方法选取2011年6月-2014年6月17例IS患者临床手术切除的致痫灶标本与6例尸检正常对照作比较,利用Real-time PCR、Western Blot和免疫组织化学检测IS致痫灶标本中CRH和促肾上腺皮质激素释放激素1型受体(Corticotropin releasing hormone receptor 1,CRHR1)的表达和细胞定位,分析其与癫痫发作的相关性。进一步检测下游蛋白激酶C(Protein kinase C,PKC)信号的表达,分析其与CRHR1及癫痫发作的关系。结果在17例IS致痫灶标本中,CRH和CRHR1的表达较对照组明显升高。CRH主要在神经元表达,CRHR1则分布于神经元、星形胶质细胞和小胶质细胞。CRH和CRHR1的表达水平与癫痫发作频率存在相关性。PKC高表达于IS致痫灶,分布于神经元、星形胶质细胞和小胶质细胞,且与CRHR1的表达水平及癫痫发作频率存在相关性。结论 CRH、CRHR1及其下游信号PKC高表达于IS致痫灶,且与痉挛性癫痫发作呈明显正相关,提示CRH信号途径可能参与了IS的发病机制,支持IS的CRH发病假说。

Objective The purpose of this study was to explore the expression of Corticotropin releasing hormone( CRH),Corticotropin releasing hormone receptor 1( CRHR1),Protein kinase C( PKC)in epileptogenic zone of Infantile spasm( IS). Methods Collected 17 cases of tissues of IS patients from operation and 6 cases of normal brain tissues from clinical autopsy during June 2011 to June 2014. Westen blot was used to detected the protein expression of CRH,CRHR1,PKC. PCR was used to exam the mRNA expression of CRH,CRHR1,PKC. Immunohistochemistry and fluorescenceimmuno assay were used to detect the expression of CRH,CRHR1,PKC. Results The mRNA expression of CRH and CRHR1 in IS group are higher than control group,and the protein expression of CRH and CRHR1 in IS group are higher than control group. CRH are slightly expressed in the controls,medium and strong expressed in IS,CRH and NF200 both expressed in IS; CRH is negative in GFAP positive astrocyte; CRH is negative in HLA positive microglial cell. CRHR1 are slightly and medium expressed in the controls,medium and strong expressed in IS,CRHR1 and NF200 both expressed in IS; CRHR1 and GFAP are both positive in astrocyte; CRHR1 and HLA are both positive in microglial cell. PKC are slightly and medium expressed in the controls,medium and strong expressed in IS,PKC and NF200 both expressed in IS; PKC and GFAP are both positive in astrocyte; PKC and HLA are both positive in microglial cell. Spearman analysis showed positive correlation between the expression of CRH,CRHR1,PKC with epileptic spasm in IS patients,as well as positive correlation between PKC with CRHR1. Conclusions Over expression of CRH,CRHR1,PKC with epileptic spasm in IS patients were positive related with epileptic seizure in IS patients,indicated that CRH signal pathway is related with IS pathogenesis.