托珠单抗对心衰大鼠辅助性T细胞17介导的心肌炎症反应的影响  

Effects of Tocilizumab on myocardial inflammation mediated by Th17 cells in rats with chronic heart failure

作　　者：张海宁[1] 尹园[1] 柳玉梅[1] 张维文[1] 

机构地区：[1]广州医科大学药理教研室,广东广州511436

出　　处：《中国现代医学杂志》2015年第35期7-12,共6页China Journal of Modern Medicine

基　　金：广州市教育局基金(No:10A179)

摘　　要：目的探讨托珠单抗对心衰大鼠辅助性T细胞17(Th17)介导的心肌炎症反应的影响及对心功能的保护作用。方法利用冠状动脉左前降支结扎法制备心衰模型,于术后第2天腹腔注射5 mg/kg托珠单抗,4周注射1次,连续注射3次。实验结束后,左心室插管检测大鼠左心室血流动力学变化;酶联免疫吸附(ELISA)法测定心肌或脾脏组织中肿瘤坏死因子-α(TNF-α)、白介素-17(IL-17)、白介素-1β(IL-1β)、白介素-6(IL-6)及白介素-23(IL-23)水平;流式细胞仪检测脾脏Th17细胞比例;实时荧光定量聚合酶链反应(RTq PCR)检测脾脏维A酸相关孤儿受体γt(RORγt)及维A酸相关孤儿受体α(RORα)m RNA的表达水平;Western blot检测核转录因子-κB(NF-κB)p65核蛋白表达水平。结果托珠单抗治疗降低心衰大鼠死亡率及心脏体重比(HW/BW)比值,改善心衰大鼠血流动力学参数,包括降低平均血压(MBP)及左心室舒张末期压(LVEDP),升高左心室舒张压最大上升和下降速率(±dp/dt)及左心室收缩压(LVSP)。模型组大鼠脾组织IL-23、IL-17蛋白表达水平、Th17细胞频率、Th17细胞活化的特征性转录因子RORγt及RORαm RNA表达水平较对照组明显升高,并且心肌IL-1β、IL-6、TNF-α含量及NF-κB p65表达水平亦显著高于对照组。与模型组比较,托珠单抗治疗使上述指标明显降低。结论托珠单抗可能通过抑制Th17细胞介导的炎症反应发挥心肌保护作用。[Objective] To observe the effect of Tocilizumab on the myocardial inflammation mediated by T helper cells 17 （Th17） in rats with chronic heart failure, and to explore the possible protective action of Tocilizumab on heart function. [Methods] Male Sprague-Dawley （SD） rats were subjected to the permanent ligation of the left anterior descending coronary artery （LAD） to induce chronic heart failure. One day later, the rats in the Tocilizumab group received an intraperitoneal injection of 5 mg/kg of Tocilizumab once every four weeks for 12 weeks. Cardiac function parameters of the rats were measured by left ventricular catheteri- zation at the end of experiments. The levels of proinflammatory cytokines such as tumor necrosis factor α （TNF-α）, interleukin 1β （IL-1β）, interleukin 6 （IL-6）, intedeukin 17 （IL-17） and interleukin 23 （IL-23） in the heart and the spleen were measured by ELISA. The mRNA expressions of retinoid related orphan receptor t （ROR t） and retinoid related orphan receptor α （ROR α） were detected by real-time quantitative PCR. Th17 cell frequency was analyzed by flow cytometry and the expression level of nuclear faetor-KB （NF-KB） p65 in the cardiomyocyte nuclei was examined using Western blot. [Results] Tocilizumab treatment significantly de- creased the rat mortality and the heart-to-body weight ratio （HW/BW）, and ameliorated rat cardiac function, as evidenced by decreasing mean blood pressure （MBP） and left ventricular end-diastolic pressure （LVEDP） and increasing the maximum ascending and descending rates of left ventricular diastolic pressure （＋ dp/dt） and left ventricular systolic pressure （LVSP）. Moreover, the rats in the model group exhibited significantly increased IL-23 and IL-17 expressions, Thl7 cell frequency, mRNA levels of RORt and RORot in the rat spleen, as well as increased cardiomyocyte NF-κB p65 expression and myocardial content of IL-I[3, IL-6 and TNF-tx as com- pared with those in the control group; but all

关 键 词：心衰 炎症 托珠单抗 NF—κB 

分 类 号：R541.6[医药卫生—心血管疾病]