视黄酸相关孤核受体α对脓毒症小鼠心肌损伤的影响

Role of retinoic acid-associated orphan nuclear receptor alpha in myocardial injury in septic mice

目的 本研究旨在探索视黄酸相关孤核受体α(RORα)对脓毒症炎症反应及其所致的心脏功能损害影响。方法 通过腹腔注射脂多糖(LPS)建立小鼠脓毒症模型,通过LPS刺激巨噬细胞,建立脓毒症细胞模型,采用细胞转染和小鼠尾静脉注射过表达质粒,以在细胞和组织中过表达RORα,进而探究RORα在脓毒症炎症反应和脓毒症所致心肌损伤中的作用。采用实时定量PCR试验、蛋白免疫印记试验检测RORα、NF-κBp65和炎症因子的变化,采用酶联免疫试验、病理切片等检测脓毒症小鼠心肌损伤的变化。结果 RORα在LPS刺激巨噬细胞中显著降低(P<0.05);过表达RORα能够显著抑制LPS刺激巨噬细胞中炎症因子白介素(IL)-1β和肿瘤坏死因子(TNF)-α的表达(P<0.05)、NF-κBp65的核移位水平(P<0.05)、脓毒症小鼠血清中炎症因子IL-1β和TNF-α的释放、降低脓毒症小鼠肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)的含量以及改善脓毒症所致的心肌病理损害。结论 RORα能够通过抑制NF-κBp65的核移位进而负性调控LPS刺激巨噬细胞中的炎症反应,从而缓解脓毒症小鼠的炎症反应及心肌损害。

AIM To investigate the role of retinoic acid-related orphan nuclear receptor alpha(RORα)in inflammatory responses and cardiac function injury to sepsis.METHODS A mouse model of sepsis was established by intraperitoneal injection of lipopolysaccharide(LPS).Macrophage was stimulated by LPS to establish a septic cell model.Cell transfection and mouse tail vein injections of over-expression plasmids were used to over-express RORαin cells and tissues and to explore the role of RORαin inflammatory responses and myocardial damage mediated by sepsis.The changes of RORα,NF-κB p65 and inflammatory cytokines were detected by real-time quantitative PCR and Western blot.The changes of cardiac function in mice with sepsis were detected by enzyme-linked immunosorbent assay and pathological section RESULTS RORαsignificantly decreased in LPS-stimulated macrophages(P<0.05).Over-expression of RORαsignificantly inhibited the expression of inflammatory cytokines IL-1βand TNF-αin LPS-stimulated macrophages(P<0.05).Over-expression of RORαsignificantly inhibited LPS stimulated nuclear translocation of NF-κB p65 in macrophages(P<0.05).Over-expression of RORαin vivo significantly inhibited the release of inflammatory factors IL-1βand TNF-αin the serum of septic mice,reduced the content of myocardial enzymes LDH and CK-MB in septic mice and improved myocardial pathological damage mediated by sepsis.CONCLUSION RORαcan inhibit the inflammatory response of macrophages by inhibiting the nuclear translocation of NF-κB p65,thereby alleviating the inflammatory response and myocardial injury in mice with sepsis.