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大鼠早期心肌缺血干燥综合征抗原B基因及其蛋白的表达

Expression of SSB Following Early Myocardial Ischemia in Rats

  • 摘要: 目的 探讨干燥综合征抗原B基因(SSB)及其蛋白在大鼠早期心肌缺血组织中的表达。方法 健康SD大鼠随机分为手术组、非手术对照组和假手术组,根据缺血时间分为0 min、15 min、30 min、60 min、120 min和240 min亚组,依据缺血部位将手术组又分为心肌缺血组和心肌非缺血组。手术组每亚组6只大鼠,非手术对照组及假手术组中的每亚组4只。通过实时荧光定量PCR测定每个样本SSB基因的表达水平,运用Western blot和免疫荧光技术检测SSB蛋白的表达。结果 SSB基因表达在急性心肌缺血早期呈下调趋势,心肌缺血120 min组SSB mRNA 表达降低,与0 min组及非手术对照组相比,差异具有统计学意义(P<0.05);与心肌缺血0 min组相比,SSB蛋白在心肌缺血60 min组的组织中的表达降低(P<0.05);心肌缺血60 min和120 min,SSB蛋白在非缺血组中的表达高于缺血组(P<0.05)。缺血心肌组织中,SSB蛋白主要位于心肌细胞核,胞浆及部分心肌细胞的细胞膜上也有少量表达。结论 SSB在急性心肌缺血后呈现差异性表达,可能参与早期缺血心肌的病理生理过程。

     

    Abstract: Objective To investigate the expression of Sj?gren’s syndrome antigen B (SSB) gene and SSB protein in the early ischemic myocardium in rats. Methods Adult healthy Sprague-Dawley rats were randomly divided into groups of operation 〔myocardial ischemia (MI) and non-ischemia (NI)〕, non-operation (NO) and sham-operation (SO) (n=6 for MI and NI; n=4 for NO and SO). According to time of ischemia, it was then divided into groups of 0 min, 15 min, 30 min, 60 min, 120 min, and 240 min. The expression of SSB gene in the myocardium was examined by real-time PCR, and the expression of SSB protein was examined by Western blot and immunofluorescence staining. Results The expressions of SSB gene was down-regulated at early stage of ischemia. There was significant difference between 0 min and 120 min at the level of expression of SSB gene in MI group, so did that between 120 min group and NO group (P<0.05). The expression of SSB protein at 60 min after ischemia was significantly decreased compared with that in the group of 0 min (P<0.05). The expression of SSB protein in NI groups was significantly higher than that in MI groups at the time of 60 min and 120 min after myocardial ischemia (P<0.05). Additionally, the expression of SSB protein was mainly located in the myocardial nucleus, myocardial plasma, and plasma membrane of partial myocardiocytes according to the result of immunofluorescence staining. Conclusion SSB may participate in pathophysiologic regulation process in myocardial cells at the early stage of myocardial ischemia in rats.

     

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