中性粒细胞弹性蛋白酶抑制剂sivelestat对HFpEF小鼠心脏纤维化的作用  

Effect of neutrophil elastase inhibitor sivelestat on cardiac fibrosis in HFpEF mice

作　　者：甘静 张克 杜函霖 林灼锋 吴震[1] GAN Jing;ZHANG Ke;DU Hanlin;LIN Zhuofeng;WU Zhen(Department of Cardiovascular Medicine,The Third Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510630,China;Center for Cardiometabolic Disease Innovation,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]中山大学附属第三医院心血管内科,广东广州510630 [2]广东医科大学心血管代谢病创新中心,广东东莞523808

出　　处：《中国病理生理杂志》2025年第1期11-18,共8页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.2020A1515010362)。

摘　　要：目的:观察中性粒细胞弹性蛋白酶(neutrophil elastase,NE)抑制剂sivelestat对射血分数保留型心力衰竭(heart failure with preserved ejection fraction,HFpEF)小鼠心脏组织病理结构变化的影响。方法:8周龄C57BL/6J小鼠随机分为对照组(n=5)、模型组(n=6)和治疗组(n=18)。采用高脂饮食加N^(G)-硝基-L-精氨酸甲酯(N^(G)nitro-L-arginine methyl ester,L-NAME)联合造模的方法建立HFpEF小鼠模型。治疗组分为3组(n=6),按照12.5、25和50 mg·kg^(-1)·d^(-1)的剂量腹腔注射sivelestat,对照组和模型组注射等体积的生理盐水,连续给药12周。使用小动物超声检测小鼠心功能的变化;计算小鼠肺脏重量(lung weight,LW)与胫骨长度(tibial length,TL)的比值(LW/TL)以评估肺水肿;进行运动疲劳测试评估运动不耐受程度;通过HE、小麦胚芽凝集素、二氢乙啶及天狼星红染色依次评估心脏整体病理变化、心肌细胞横截面积、氧化应激水平及心脏纤维化面积;通过Western blot检测纤维化相关蛋白的表达。结果:与模型组比较,不同浓度的sivelestat可显著改善HFpEF小鼠心脏舒张功能,表现为E波/A波比值(E/A)增加和E波/e'波比值(E/e')降低(P<0.05);LW/TL降低,肺充血及运动不耐受减轻(P<0.05);心脏重量(heart weight,HW)与TL的比值(HW/TL)降低,心脏整体和心肌细胞横截面积减小,心脏肥大显著减轻(P<0.05);此外,心肌纤维化及活性氧水平也显著降低(P<0.05)。结论:抑制NE可抑制HFpEF引起的小鼠心脏舒张功能障碍和心脏纤维化,从而对心脏具有保护效应。AIM:To investigate the effects of the neutrophil elastase(NE)inhibitor sivelestat on the pathological changes of cardiac tissues in mice with heart failure with preserved ejection fraction(HFpEF).METHODS:Eightweek-old C57BL/6J mice were randomly divided into 3 groups:control group(n=5),model group(n=6),and treatment group(n=18).The HFpEF mouse model was established using a combined approach of high-fat diet and N^(G)-nitro-L-arginine methyl ester(L-NAME).The mice in treatment group received intraperitoneal injection of sivelestat at doses of 12.5,25 and 50 mg·kg^(-1)·d^(-1)(n=6),while those in control and model groups were injected with the same volume of normal saline.After 12 weeks,small animal ultrasound was employed to assess changes in cardiac function,and the lung weight-to-tibia length(LW/TL)ratio was calculated to evaluate pulmonary edema.An exercise fatigue test was conducted to assess exercise intolerance.Histological evaluations were performed using HE,wheat germ agglutinin,dihydroethidium and Sirius red staining to examine overall heart morphology,cross-sectional area of cardiomyocytes,levels of oxidative stress,and the extent of cardiac fibrosis.The expression of fibrosis-related proteins was analyzed using Western blot.RE⁃SULTS:Compared with model group,sivelestat at various concentrations significantly improved cardiac diastolic function in HFpEF mice,indicated by an increased E/A ratio and a decreased E/e'ratio(P<0.05).The LW/TL ratio decreased,alleviating lung congestion and exercise intolerance(P<0.05).The heart weight-to-tibia length(HW/TL)ratio,overall heart cross-sectional area,and cardiomyocyte cross-sectional area all decreased,indicating attenuation in cardiac hypertrophy(P<0.05).Additionally,cardiac fibrosis and reactive oxygen species levels were significantly reduced(P<0.05).CONCLUSION:Inhibition of NE exerts a protective effect against diastolic dysfunction and cardiac fibrosis induced by HFpEF in mice.

关 键 词：中性粒细胞弹性蛋白酶 SIVELESTAT 射血分数保留型心力衰竭 心脏纤维化 

分 类 号：R541[医药卫生—心血管疾病] R363.2[医药卫生—内科学] Q291[医药卫生—临床医学]