CAJ | 학술논문

目的探讨葡萄糖调节蛋白78(GRP78)、p38丝裂素激活蛋白激酶(p38MAPK)相关信号通路在惊厥性脑损伤中的作用及尼莫地平对其的影响.方法雄性SD幼年大鼠分为惊厥持续状态组(SC组)、尼莫地平组(NM组)、正常对照组(NC组).采用免疫组织化学法、反转录(RT)-PCR技术检测海马CA1区GRP78/Bip、p38MAPK蛋白、mRNA表达动态变化;采用原位末端标记(TUNEL)检测神经细胞的凋亡.结果 1.免疫组织化学:GRP78/Bip蛋白惊厥后4h开始增加,24 h达高峰,之后下降.p38MAPK蛋白惊厥后4h表达开始增加,24 h达高峰,48 h稍有下降.2.RT-PCR:SC组海马GRP78/Bip mRNA表达于惊厥后4h开始少量增加,24 h达高峰,48 h回到基线水平.NM组4h,24 h和48 h表达情况较SC组和NC组均明显升高(P均＜0.05).SC组p38MAPK mRNA于惊厥后4h开始表达,24 h达高峰,48 h后有所下降,但均高于NC组;NM组24 h时间点明显高于NC组(P＜0.01).3.TUNEL:SC组海马CA1区TUNEL阳性细胞于惊厥后4h已有少量表达,48 h达高峰,之后有下降趋势;且24 h、48 h2个时间点均明显高于NC组(P均＜0.05).NM组24 h、48 h时间点TUNEL阳性细胞表达水平较SC组明显降低(P均＜0.05);但仍高于NC组(P＜0.01).结论 GRP78信号通路可能通过激活p38MAPK介导细胞凋亡,尼莫地平预处理可影响GRP78/Bip和p38MAPK的表达,缓解内质网应激,减轻惊厥后海马病理损伤程度.
목적 탐토포도당조절단백78(GRP78)、p38사렬소격활단백격매(p38MAPK)상관신호통로재량궐성뇌손상중적작용급니막지평대기적영향.방법 웅성SD유년대서분위량궐지속상태조(SC조)、니막지평조(NM조)、정상대조조(NC조).채용면역조직화학법、반전록(RT)-PCR기술검측해마CA1구GRP78/Bip、p38MAPK단백、mRNA표체동태변화;채용원위말단표기(TUNEL)검측신경세포적조망.결과 1.면역조직화학:GRP78/Bip단백량궐후4h개시증가,24 h체고봉,지후하강.p38MAPK단백량궐후4h표체개시증가,24 h체고봉,48 h초유하강.2.RT-PCR:SC조해마GRP78/Bip mRNA표체우량궐후4h개시소량증가,24 h체고봉,48 h회도기선수평.NM조4h,24 h화48 h표체정황교SC조화NC조균명현승고(P균＜0.05).SC조p38MAPK mRNA우량궐후4h개시표체,24 h체고봉,48 h후유소하강,단균고우NC조;NM조24 h시간점명현고우NC조(P＜0.01).3.TUNEL:SC조해마CA1구TUNEL양성세포우량궐후4h이유소량표체,48 h체고봉,지후유하강추세;차24 h、48 h2개시간점균명현고우NC조(P균＜0.05).NM조24 h、48 h시간점TUNEL양성세포표체수평교SC조명현강저(P균＜0.05);단잉고우NC조(P＜0.01).결론 GRP78신호통로가능통과격활p38MAPK개도세포조망,니막지평예처리가영향GRP78/Bip화p38MAPK적표체,완해내질망응격,감경량궐후해마병리손상정도.
Objective To explore the role of glucose-regulated protein 78 (GRP78),p38 mitogen-activated protein kinase(p38MAPK) signal pathway in seizure-reduced brain injures and the regulatory effect of Nimodipine on it.Methods Sprague-Dawley(SD) rats were randomly divided into status convulsion group (SC group),Nimodipine group(NM group),and a normal control group(NC group).The expressions of GRP78/Bip and p38MAPK mRNA and protein were detected by reverse transcription(RT)-PCR and immunohistochemistry.The expression of apoptosis cells was observed by TdT-mediated dUTP nick end labeling (TUNEL).Results (1) Immunohistochemistry:at 4 h after induction of status convulsion,the expression of GRP78 protein in the hippocampus CA1 domain began increasing slightly,and reached a maximum at 24 h,and then began decreasing slowly ; at 4 h after induction of status convulsion,the expression of p38MAPK protein in the hippocampus CA1 domain began increasing,and reached a maximum at 24 h,and decreased remarkably at 48 h.(2) RT-PCR:at 4 h after induction of status convulsion,the expression of GRP78 protein in the hippocampus CA1 domain began increasing slightly,and reached a maximum at 24 h,and then began decreasing slowly.The NM group was much higher than the SC group and the NC group(all P ＜ 0.05) ; at 4 h after induction of status convulsion,the expression of p38MAPK protein in the hippocampus CA1 domain began increasing,and reached a maximum at 24 h,and decreased remarkably at 48 h;the NM group was much lower than the SC group,and higher than the NC group (all P ＜ 0.05).(3) TUNEL:at 4 h after induction of status convulsion,the expression of the TUNEL positive cells in the hippocampus CA1 domain began increasing slightly,and reached a maximum at 48 h,and then began decreasing,and there was no difference between SC group and NC group;the NM group was much lower than the SC group(all P ＜ 0.05).Conclusions The correlation of the increased expression of p38MAPK and neuronal apoptosis indicates that GRP78 signal pathway may be mediated to cell apoptosis through p38MAPK.Nimodipine can affect the expression of GRP78/Bip and p38MAPK,and relieve endoplasmic reticulum stress,and lessen the pathologic damage to the hippocampus.