Shafi Shahida, Codrington Rosalind, Gidden Lewis Michael, Ferns Gordon Ashley Anthony
Faculty of Health and Medical Sciences, Department of Biochemistry and Physiology, University of Surrey, Guildford, Surrey, UK.
Babraham Research Campus, ABeterno Ltd, Cambridge, UK.
Int J Exp Pathol. 2016 Feb;97(1):56-65. doi: 10.1111/iep.12167. Epub 2016 Feb 8.
We aimed to assess the expression and distribution of Hsp27, pHsp27 (Ser82), p38MAPK and p-p38MAPK in fibro-fatty atherosclerotic lesions and the myocardium of hypercholesterolaemic rabbits. Male New Zealand white rabbits were fed a high-cholesterol diet for 18 weeks, maintaining serum cholesterol at approximately 20 mmol/l over this period. Aortic arch and myocardial tissues were analysed by Western blot, immunohistochemistry and double immunofluorescence. Plasma Hsp27 levels were measured by ELISA. There was a significant increase in the expression of monomeric and dimeric forms of Hsp27, together with pHsp27 (Ser82), p38MAPK and p-p38MAPK in the fibro-fatty atherosclerotic lesions (P < 0.01; P < 0.05; P < 0.001; and P < 0.001, respectively) and the myocardial tissues (P < 0.001) from the cholesterol-fed rabbits compared with equivalent tissues from controls when the plasma concentration was low. Immunohistochemical analysis of the fibro-fatty lesions showed marked increases in Hsp27 and pHsp27 (Ser82) immunoreactivity. Double immunostaining showed intense expression of pHsp27 and p-p38MAPK in regions that were rich in macrophages, suggesting a close association with these inflammatory cells, whereas, in regions rich in smooth muscle cells, only p-p38MAPK was found to be strongly expressed. An increased expression of pHsp27 (Ser82) was spatially associated with increased p-p38MAPK within fibro-fatty atherosclerotic lesions and was colocalized to regions rich in macrophages. The initial increase in plasma Hsp27 levels may reflect the increase in systemic inflammation and oxidative stress in the early phases of disease. The falling concentrations subsequently may be coincident with the development of the advanced atherosclerotic lesions.
我们旨在评估热休克蛋白27（Hsp27）、磷酸化热休克蛋白27（Ser82位点）、p38丝裂原活化蛋白激酶（p38MAPK）和磷酸化p38丝裂原活化蛋白激酶（p-p38MAPK）在高胆固醇血症兔的纤维脂肪粥样硬化病变及心肌中的表达和分布情况。雄性新西兰白兔接受高胆固醇饮食喂养18周，在此期间将血清胆固醇维持在约20 mmol/L。通过蛋白质免疫印迹法、免疫组织化学和双重免疫荧光法对主动脉弓和心肌组织进行分析。采用酶联免疫吸附测定法（ELISA）检测血浆Hsp27水平。与血浆浓度较低时对照组的相应组织相比，在胆固醇喂养兔的纤维脂肪粥样硬化病变（分别为P < 0.01；P < 0.05；P < 0.001；和P < 0.001）及心肌组织（P < 0.001）中，Hsp27的单体和二聚体形式、磷酸化热休克蛋白27（Ser82位点）、p38MAPK和磷酸化p38MAPK的表达均显著增加。对纤维脂肪病变进行免疫组织化学分析显示，Hsp27和磷酸化热休克蛋白27（Ser82位点）的免疫反应性显著增加。双重免疫染色显示，在富含巨噬细胞的区域，磷酸化热休克蛋白27和磷酸化p38MAPK表达强烈，提示与这些炎性细胞密切相关；而在富含平滑肌细胞的区域，仅发现磷酸化p38MAPK强烈表达。在纤维脂肪粥样硬化病变内，磷酸化热休克蛋白27（Ser82位点）表达增加在空间上与磷酸化p38MAPK增加相关，并定位于富含巨噬细胞的区域。血浆Hsp27水平最初升高可能反映了疾病早期全身炎症和氧化应激的增加。随后浓度下降可能与晚期动脉粥样硬化病变的发展同时出现。
