分泌型酸性富含半胱氨酸样蛋白1型和整合素β1在致痫大鼠中的表达变化及相关性分析  

Expression change and correlation analysis of SPARCL1 and Integrinβ1 in epileptic rats

作　　者：劳基通 邝希 刘惠 肖敏[1] 陈永敏[1] 廖小平[1] 马琳[1] 李其富[1] LAO Jitong;KUANG Xi;LIU Hui;XIAO Min;CHEN Yongmin;LIAO Xiaoping;MA Lin;LI Qifu(不详;Department of Neurology,First Affiliated Hospital of Hainan Medical College,Haikou Hainan 570100,China)

机构地区：[1]海南医学院第一附属医院神经内科,571300

出　　处：《中国神经免疫学和神经病学杂志》2022年第2期123-129,共7页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金(81760244,81960249);海南省重点研发科技计划(ZDYF2017164,ZDYF2019124);海南省临床医学中心建设项目资助(2021)。

摘　　要：目的研究癫痫大鼠海马和邻近颞叶皮层中分泌型酸性富含半胱氨酸样蛋白1型(secreted protein acidic and rich in cysteine1,SPARCL1)和整合素β1(Integrinβ1)的动态表达,以探讨二者对癫痫发展的影响。方法将雄性SD大鼠56只随机分为对照组(21只)和致痫组(35只)。致痫组予氯化锂(LiCl)匹罗卡品(PILO)进行造模,按癫痫发作后3 h、6 h、12 h、1 d、2 d、3 d、7 d时间点分为7个亚组,每亚组5只;对照组按对应时间分为7个亚组,每亚组3只。采用实时荧光定量PCR(qPCR)检测大鼠SPARCL1mRNA和Integrinβ1 mRNA表达水平,蛋白免疫印迹杂交实验(Western blot)检测大鼠SPARCL1和Integrinβ1的蛋白表达水平。比较致痫组和对照组大鼠海马组织和颞叶皮层组织中SPARCL1和Integrinβ1的mRNA和蛋白表达的变化,并分析大鼠癫痫发作后不同时间不同组织中SPARCL1和Integrinβ1的mRNA和蛋白表达水平的相关性。结果对照组7个时间亚组的海马组织和颞叶皮层样本中SPARCL1和Integrinβ1的mRNA和蛋白表达水平差异均无统计学意义(均P<0.05)。与对照组比较,致痫组3 h组、6 h组、12 h组和1 d组海马组织中SPARCL1 mRNA表达水平均降低(P<0.05),致痫组6 h组、12 h组、1 d组和2 d组海马组织中SPARCL1蛋白水平均降低(均P<0.05);致痫组海马组织3 h组和6 h组的Integrinβ1 mRNA表达水平明显下降(P<0.01),致痫组1 d组和2 d组Integrinβ1 mRNA表达水平显著增加(P<0.01);致痫组6 h、12 h组的Integrinβ1蛋白表达水平下降(P<0.05),3 d、7 d组Integrinβ1蛋白表达水平升高(均P<0.05)。与对照组比较,致痫组3 h组、6 h组和7 d组大鼠颞叶皮层中SPARCL1 mRNA表达水平显著升高(P<0.01),致痫组12 h组和1 d组则明显下降(P<0.01),致痫组12 h组、1 d组、2 d组、3 d组和7 d组SPARCL1蛋白表达水平显著降低(均P<0.05);致痫组3 h组、6 h组和7 d组大鼠颞叶皮层Integrinβ1的mRNA显著增加(P<0.01),除致痫组3 h组外,致痫组6 h组、12 h组、1 d组�Objective To study the dynamic expression of secreted protein acidic and rich in cysteine-like protein type 1(SPARCL1)and integrinβ1 in the hippocampus and adjacent temporal cortex of epileptic rats for investigating their effects on the development of epilepsy.Methods Fifty-six male SD rats were randomly divided into a control group(21 rats)and an epileptogenic group(35 rats).The epileptogenic group was modeled with lithium chloride(LiCl)and pilocarpine(PILO)and the epileptogenic group was divided into 7 subgroups according to the observation time(3 h,6 h,12 h,1 d,2 d,3 d and 7d)after seizure,with 5 animals in each subgroup.The control group was divided into 7 subgroups according to the corresponding time with 3 rats in each subgroup.Real-time fluorescence quantitative PCR(qPCR)was used to detect the mRNA expression levels of both SPARCL1 and integrinβ1 transcripts and protein immunoblot hybridization assay(western blot)was used to detect the protein expression levels of SPARCL1 and integrinβ1 translations in the hippocampus and adjacent temporal cortex of the rats.The correlation between the mRNA and protein expression of SPARCL1 and integrinβ1 in different tissue at different time points after seizures in the rats was analyzed.Results The differences in mRNA and protein expression levels of SPARCL1 and integrinβ1 in hippocampal tissue and temporal cortex samples from the seven temporal subgroups of the control group were not statistically significant(all P<0.05).Compared with control group,SPARCL1 mRNA expression levels were reduced in hippocampal tissue in the 3 h,6 h,12 h and 1 d epileptogenic groups(P<0.05),and SPARCL1 protein levels were reduced in hippocampal tissue in the 6 h,12 h,1 d and 2 d epileptogenic groups(all P<0.05).Compared with the control group,SPARCL1 mRNA expression levels were significantly higher in the temporal cortex of the rats in the 3 h,6 h and 7 d epileptogenic groups(P<0.01),significantly lower in the 12 h and 1 d epileptogenic groups(P<0.01).SPARCL1 protein expression levels

关 键 词：癫痫 分泌型酸性富含半胱氨酸样蛋白1型 整合素Β1 

分 类 号：R742.1[医药卫生—神经病学与精神病学]