摘要
目的探索黄芪甲苷(AS-Ⅳ)通过PINK1/Parkin信号通路调控自噬减轻5-氟尿嘧啶(5-Fu)诱导老龄大鼠心肌毒性的疗效及机制。方法18月龄雄性SD大鼠36只,按随机数字表法分为对照组、模型组、AS-Ⅳ组,每组12只。以5-Fu注射液30 mg/kg,腹腔注射,隔日1次,连续7次建立模型组;AS-Ⅳ稀释液,20 mg/kg,灌胃,每日1次,连续14次,建立AS-Ⅳ组。检测血清肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)、心肌线粒体腺苷三磷酸(ATP)及膜电位水平、行HE及Masson染色观察心肌病理改变、免疫荧光染色检测微管相关蛋白LC3Ⅱ表达、电镜观察线粒体结构并采用Western blot检测PINK1/Parkin通路关键分子PINK1、Parkin、Beclin1、LAMP、LC3蛋白表达。结果与模型组相比,AS-Ⅳ可减缓大鼠体质量下降,降低心肌cTnI及CK-MB水平,抑制线粒体ATP水平下降(P<0.05)。与对照组相比,模型组心肌纤维排列紊乱,胶原纤维沉积,胶原容积百分比升高;经AS-Ⅳ干预后,胶原容积百分比减低(P<0.05)。电镜及LC3Ⅱ荧光染色显示,经AS-Ⅳ干预后,线粒体损伤程度减轻,LC3Ⅱ表达量减低(P<0.05)。Western blot结果提示AS-Ⅳ苷干预后Beclin1、PINK1、Parkin、LAMP和LC3Ⅱ/Ⅰ表达较模型组降低(P<0.05)。结论5-Fu可诱导心肌线粒体损伤,线粒体自噬过度,AS-Ⅳ可通过调节自噬减轻5-Fu心肌毒性。
Objective To investigate the effect and mechanism of astragaloside Ⅳ(AS-Ⅳ) regulating autophagy to reduce 5-fluorouracil(5-Fu) induced myocardial toxicity based on PINK1/Parkin signaling pathway in the rats.Methods Thirty-six male SD rats aged 18 months were divided into control group,model group and AS-Ⅳ group by random number table method,with 12 rats in each group.The rat model was administered the intraperitoneal injection of 30 mg/kg 5-Fu every other day for 7 times.The intervention group was administered the intragastrical 20 mg/kg AS-Ⅳ diluent every day for14 times.The serum levels of troponin Ⅰ(cTnI),creatine kinase isoenzyme(CK-MB),myocardial mitochondrial ATP and membrane potential were measured.HE and Masson staining were used to evaluate the pathological changes of myocardium.The expression of microtubule-associated protein(LC3Ⅱ) was evaluated by immunofluorescence.The ultrastructure of mitochondria was observed under electron microscope.The protein expressions of PINK1/Parkin pathway(key molecules PINK 1,Parkin,Beclinl,LAMP and LC3) were detected by Western blot assay.Results Compared with the model group,AS-Ⅳ could slow down the weight loss(P <0.05),reduce the secretion of myocardial enzyme cTnI and CK-MB(P <0.05)and inhibit the decrease of mitochondrial ATP levels(P <0.05).Compared with control group,the myocardial fibers were disorderly arranged and a large number of blue collagen fibers were deposited in the model group.After the intervention of AS-Ⅳ,the collagen volume fraction decreased in the model group(P <0.05).Electron microscopy and LC3Ⅱ fluorescence labeling showed that after AS-Ⅳ intervention,the degree of mitochondrial damage was reduced and the expression of LC3Ⅱ was decreased(P<0.05).Western blot results indicated that Beclinl,PINK1,Parkin,LAMP and LC3Ⅱ/Ⅰ expression levels were significantly reduced(P <0.05).Conclusion The 5-Fu induces myocardial mitochondrial injury and excessive cardiac mitochondrial autophagy,and AS-Ⅳ alleviates it by regulating autophag
作者
李沅洋
周湘忠
雷向红
张宇凡
徐月
魏丽萍
LI Yuan-yang;ZHOU Xiang-zhong;LEI Xiang-hong;ZHANG Yu-fan;XU Yue;WEI Li-ping(Graduate School of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;Department of Cardiology,Tianjin Union Medical Center;Department of Cardiology,Tianjin Binhai New Area Dagang Hospital;Department of Ultrasound,Tianjin Union Medical Center;Graduate School of Tianjin Medical University)
出处
《天津医药》
CAS
北大核心
2021年第4期378-384,共7页
Tianjin Medical Journal
基金
天津滨海新区卫生健康委员会科技重点支持项目(2019BWKZ004)
京津冀基础研究合作专项(19JCZDC63900)。
