白藜芦醇调节RIP1/RIP3/MLKL通路对扩张型心肌病小鼠心功能的影响

Effect of resveratrol on cardiac function in mice with dilated cardiomyopathy by adjusting the RIP1/RIP3/MLKL pathway

关键词:  白藜芦醇  RIP1/RIP3/MLKL通路  扩张型心肌病  心功能

Key words:  resveratrol  RIP1/RIP3/MLKL pathway  dilated cardiomyopathy  cardiac function

摘要:

[目的] 探究白藜芦醇（RSV）调节受体相互作用丝氨酸/苏氨酸蛋白激酶（RIP）1/RIP3/混合谱系激酶域样蛋白（MLKL）通路对扩张型心肌病（DCM）小鼠心功能的影响。[方法] 雄性C57BL/6J小鼠注射阿霉素诱导DCM模型。小鼠随机分为对照组、DCM组、DCM+RSV组、DCM+肿瘤坏死因子-α（TNF-α）组、DCM+RSV+TNF-α组。超声心动图评估各组小鼠心功能；酶联免疫吸附测定（ELISA）检测各组小鼠血清心肌损伤指标；苏木精-伊红（HE）、Masson、DAPI-碘化丙啶（PI）染色观察小鼠心肌病理损伤；免疫组织化学检测各组小鼠心肌纤维化相关蛋白情况；蛋白免疫印迹法（Western blot）检测各组小鼠RIP1/ RIP3/MLKL通路相关蛋白表达。[结果] 与对照组比较，DCM组小鼠心脏质量、心脏质量/体质量比、左心室舒张末期内径（LVEDD）、肌酸激酶同工酶MB（CK-MB）、心肌肌钙蛋白T（cTn-T）、B型利钠肽（BNP）、单核细胞趋化蛋白-1（MCP-1）、心脏组织中纤维组织沉积、胶原蛋白沉积、PI阳性率、α-平滑肌肌动蛋白（α-SMA）、Ⅲ型胶原蛋白（CollagenⅢ）、TNF-α、p-RIP1/RIP1、p-RIP3/RIP3和p-MLKL/MLKL升高，左心室短轴缩短率（LVSF）、左心室射血分数（LVEF）、血红素加氧酶-1（HO-1）降低（P<0.05）。与DCM组或DCM+RSV+TNF-α组比较，DCM+RSV组小鼠心脏质量、心脏质量/体质量比、LVEDD、CK-MB、cTn-T、BNP、MCP-1、心脏组织中纤维组织沉积、胶原蛋白沉积、PI阳性率、α-SMA、Collagen Ⅲ、TNF-α、p-RIP1/RIP1、p-RIP3/RIP3和p-MLKL/MLKL降低，LVSF、LVEF、HO-1升高（P<0.05）；DCM+TNF-α组小鼠心脏质量、心脏质量/体质量比、LVEDD、CK-MB、cTn-T、BNP、MCP-1、心脏组织中纤维组织沉积、胶原蛋白沉积、PI阳性率、α-SMA、Collagen Ⅲ、TNF-α、p-RIP1/RIP1、p-RIP3/RIP3和p-MLKL/MLKL升高，LVSF、LVEF、HO-1降低（P<0.05）。[结论] RSV抑制RIP1/RIP3/MLKL通路有效改善DCM小鼠心肌纤维化，改善心脏功能。

Abstract:

[Objective] To discuss the effect of resveratrol（RSV） on cardiac function in mice with dilated cardiomyopathy（DCM） by adjusting the Receptor-Interacting Serine/Threonine-Protein Kinase（RIP）1/RIP3/Mixed Lineage Kinase Domain-Like Pseudokinase（MLKL） pathway. [Methods] Male C57BL/6J mice were injected with doxorubicin to induce DCM model. Mice were stochastically separated into Control group，DCM group，DCM+RSV group，DCM+Tumor Necrosis Factor-alpha（TNF-α） group，and DCM+RSV+TNF-α group. Echocardiography was used to evaluate the cardiac function of mice in each group. ELISA was used to detect serum myocardial injury indicators of mice. HE，Masson，and DAPI propidium iodide（PI） stainings were used to observe pathological myocardial injury in mice. Immunohistochemistry was used to measure fibrosis related proteins in the myocardium of mice in each group. In addition，Western blot was used to measure the RIP1/RIP3/MLKL pathway related proteins of mice. [Results] Compared with the Control group，the DCM group showed increases in mouse heart weight，heart weight/body weight ratio，left ventricular end-diastolic diameter（LVEDD），creatine kinase-myocardial band（CK-MB），cardiac troponin t（cTn-T），B-type Natriuretic Peptide（BNP），monocyte chemoattractant protein-1（MCP-1），fibrous tissue deposition，collagen deposition，PI positivity rate，alpha-smooth muscle actin（α-SMA），collagen type Ⅲ（Collagen Ⅲ），TNF-α，p-RIP1/RIP1，p-RIP3/RIP3，and p-MLKL/MLKL，and decreases in left ventricular shortening fraction（LVSF），left ventricular ejection fraction（LVEF），and heme oxygenase-1（HO-1）（P<0.05）. Compared with the DCM group or DCM+RSV+TNF-α group，the DCM+RSV group showed decreases in mouse heart weight，heart weight/body weight ratio，LVEDD，CK-MB，cTn-T，BNP，MCP-1，fibrous tissue deposition，collagen deposition，PI positivity rate，α-SMA，Collagen Ⅲ，TNF-α，p-RIP1/RIP1，p-RIP3/RIP3，and p-MLKL/MLKL，and increases in LVSF，LVEF，and HO-1（P<0.05）；the DCM+TNF-α group showed increases in mouse heart weight，heart weight/body weight ratio，LVEDD，CK-MB，cTn-T，BNP，MCP-1，fibrous tissue deposition，collagen deposition，PI positivity rate，α-SMA，Collagen Ⅲ，TNF-α，p-RIP1/RIP1，p-RIP3/RIP3，and p-MLKL/MLKL，and decreases in LVSF，LVEF，and HO-1（P<0.05）. [Conclusion] RSV effectively improves myocardial fibrosis and enhances cardiac function in DCM mice by inhibiting the RIP1/RIP3/MLKL pathway.