2024 年第 9 期 第 0 卷

一种新型溴结构域和超末端结构域小分子抑制剂干预小鼠心力衰竭的作用

Effects of a novel small molecule inhibitor of the bromodomain and extra-terminal domain on intervention of heart failure in mice

作者：成晨朱岳鑫王雪李凤娟王媛

英文作者：Cheng Chen Zhu Yuexin Wang Xue Li Fengjuan Wang Yuan

单位：首都医科大学附属北京安贞医院北京市心肺血管疾病研究所心血管生物研究室教育部重塑相关心血管疾病重点实验室心血管重大疾病防治协同创新中心，北京100029

英文单位：Beijing Anzhen Hospital Capital Medical University Cardiovascular Biology Laboratory Beijing Institute of Heart Lung and Blood Vessel Diseases Key Laboratory of Remodeling-related Cardiovascular Diseases Ministry of Education Collaborative Innovation Center for Cardiovascular Disorders Beijing 100029 China

关键词：心力衰竭；心脏纤维化；表观遗传修饰；溴结构域和超末端结构域

英文关键词：Heartfailure；Cardiacfibrosis；Epigeneticmodification；Bromodomainandextra-terminaldomain

• 摘要：

• 目的 探讨一种新型溴结构域和超末端结构域（BET）小分子抑制剂ABBV-744在干预心力衰竭小鼠心脏纤维化中的作用。方法 将24只8～10周龄雄性C57BL/6J小鼠随机分为假手术组、模型组、JQ1干预组和ABBV-744干预组，每组6只。假手术组不建立心力衰竭模型，其余3组小鼠行主动脉缩窄术（TAC）构建心力衰竭模型，通过检测主动脉弓流速确定造模成功。在TAC术后18 d开始2个干预组小鼠分别进行JQ1腹腔注射和ABBV-744灌胃，1次/d，连续给药30 d。给药结束后，采用超声心动图检测小鼠心功能情况；心脏相关指数分析小鼠心脏肥大情况；苏木精-伊红（HE）染色分析小鼠心脏以及肾脏组织炎症细胞浸润情况；天狼星红染色评估小鼠心脏组织纤维化程度；蛋白质印迹法和荧光定量聚合酶链反应验证小鼠心脏组织α-平滑肌肌动蛋白（α-SMA）、α1-Ⅰ型胶原（COL1A1）、α1-Ⅲ型胶原（COL3A1）表达情况；取血浆检测天冬氨酸转氨酶（AST）、总胆红素、血肌酐、血尿素氮指标评估药物对肝肾功能的影响。结果 超声心动图显示与模型组比较，ABBV-744干预组小鼠的左心室射血分数和左心室短轴缩短率均显著升高［（55±4）%比（45±4）%、（28.0±2.6）%比（22.4±2.4）%］，左心室收缩末期内径和左心室舒张末期内径均显著降低(P＜0.05或P＜0.01)。与模型组比较，ABBV-744干预组小鼠心脏重量与体重比值、心脏重量与胫骨长度比值均明显减小（P＜0.05或P＜0.001）。HE染色表明ABBV-744减轻了心脏微血管的炎症细胞浸润；天狼星红染色表明ABBV-744能够减轻TAC后心力衰竭的胶原沉积，减轻心脏纤维化。与模型组比较，ABBV-744干预组α-SMA蛋白和COL1A1、COL3A1 mRNA表达均明显下调(P＜0.05或P＜0.01)。小鼠肾脏HE染色和血浆生化检测提示与JQ1相比，ABBV-744对小鼠肝肾损伤更轻。结论 ABBV-744能有效改善TAC小鼠心力衰竭并减轻心脏纤维化，可能与抑制成纤维细胞激活蛋白相关，与JQ1相比对肝肾功能影响更小。

• Objective To investigate the effect of ABBV-744, a novel small molecule inhibitor of the bromodomain and extra-terminal domain (ABBV-744) on cardiac fibrosis in mice with heart failure. Methods Twenty-four male C57BL/6J mice aged 8-10 weeks were randomly divided into sham-operated group, model group, JQ1 intervention group and ABBV-744 intervention group, with 6 mice in each group.Heart failure model was not established in the sham operation group, and the heart failure model was established by transverse aortic constriction (TAC) in the other three groups. The success of the model was determined by detecting the flow velocity of the aortic arch. After 18 d TAC surgery, JQ1 intervention group were administered by intraperitoneal injection, and ABBV-744 was administered by gavage once a day for 30 d. Cardiac function was detected by echocardiography. Cardiac hypertrophy was analyzed by cardiac related index. Hematoxylin eosin (HE) staining was used to analyze the infiltration of inflammatory cells in the heart and kidney. Sirius red staining was used to evaluate the degree of cardiac fibrosis in mouse heart tissue. Western blotting and quantitative polymerase chain reaction were used to detect the expression of α-smooth muscle actin(α-SMA), type α1-Ⅰ collagen (COL1A1), and type α1-Ⅲ collagen (COL3A1). Aspartate aminotransferase, total bilirubin, serum creatinine and blood urea nitrogen were detected to evaluate the effect of drugs on liver and kidney function. Results Echocardiography showed that the left ventricular ejection fraction and fractional shortening were significantly increased in the ABBV-744 intervention group as compared with the model group［(55±4)% vs (45±4)%, (28.0±2.6)% vs (22.4±2.4)%］, and both left ventricular endsystolic diameter and left ventricular enddiastolic diameter were significantly reduced (P<0.05 or P<0.01). Compared with the model group, the ratio of heart weight to body weight and the ratio of heart weight to tibia length were significantly decreased in the ABBV-744 intervention group (P<0.05 or P<0.001). HE staining showed that ABBV-744 reduced the infiltration of inflammatory cells in cardiac microvessels.Sirius red staining showed that ABBV-744 could reduce collagen deposition and cardiac fibrosis in heart failure after TAC.Compared with the model group, the protein expression of α-SMA and the mRNA expression of COL1A1 and COL3A1 in the ABBV-744 intervention group was significantly down-regulated(P<0.05 or P<0.01). Compared with the JQ1 intervention group,HE staining of the kidney and biochemical analysis of the plasma showed that ABBV-744 had less damage to the liver and kidney. Conclusion ABBV-744 can effectively improve TAC mice heart failure and reduce cardiac fibrosis, which may be related to the inhibition of fibroblast activation protein. Compared with JQ1 intervention group, ABBV-744 has less effect on liver and kidney function.