D-核糖对阿霉素诱导心脏毒性的保护作用及机制研究

Protective Effect and Mechanism of D-ribose on Doxorubicin-induced Cardiotoxicity

目的:研究D-核糖对阿霉素(Doxorubicin,DOX)诱导的小鼠心脏毒性保护作用及机制。方法:8周龄雄性ICR小鼠随机分为对照组(Con),模型组(DOX)组,D-核糖低剂量组(LDR),D-核糖高剂量组(HDR),每组10只。采用单次腹腔注射大剂量阿霉素(15 mg/kg)建立DOX急性心脏毒性小鼠模型,检测血清中乳酸脱氢酶(lactate dehydrogenase,LDH)活性和心脏组织三磷酸腺苷(adenosine triphosphate,ATP)含量;通过苏木素-伊红(hematoxylin-eosin stain,HE)染色观察心肌组织病理变化;通过检测心肌组织内总超氧化物歧化酶(Total superoxide dismutase,T-SOD)、过氧化氢酶(Catalase,CAT)活性以及丙二醛(Malondialdehyde,MDA)含量,评价心脏氧化应激水平;采用蛋白免疫印迹法(Western blot)检测沉默信息调节因子2相关酶类1(silent mating type information regulation 2 homolog 1,Sirt1)、过氧化物酶体增殖物激活受体γ共激活因子1α(peroxisome proliferator-activated receptor γ coactivator-1α,PGC-1α)、B淋巴细胞瘤-2(B-cell lymphoma 2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、半胱氨酸天冬氨酸蛋白酶3(cysteine-containing aspartate specific protease 3,Caspase-3)的表达。结果:DOX可引起小鼠体质量显著降低(P<0.05),血清LDH活性显著升高(P<0.05),心肌抗氧化酶活力显著降低(P<0.05);与DOX组相比,高剂量D-核糖可以显著降低血清LDH水平(P<0.05),提高心肌抗氧化酶活力(P<0.05),提高Sirt1、PGC-1α、Bcl-2蛋白表达水平(P<0.05),降低Caspase-3、Bax蛋白表达水平(P<0.05)。结论:高剂量D-核糖可以通过激活Sirt1/PGC-1α通路,缓解氧化应激,抑制心肌细胞凋亡,降低阿霉素诱导的急性心脏毒性。

Objective: To study the protective effect and mechanism of D-ribose on Doxorubicin(DOX)-induced cardiotoxicity in mice. Methods: Eight-week-old male ICR mice were randomly divided into normal group(Con), model(DOX)group, D-ribose low-dose group(LDR) and D-ribose high-dose group(HDR), with 10 mice in each group. DOX acute cardiotoxicity mouse model was established by a single intraperitoneal injection of high dose doxorubicin(15 mg/kg).Lactate dehydrogenase(LDH) activity levels in serum and adenosine triphosphate(ATP) content in heart tissue were detected by the commercial kits. The pathological changes of myocardial tissue were observed by hematoxylin-eosin stain(HE) staining. Cardiac oxidative stress was assessed by measuring the activities of total superoxide dismutase(T-SOD),catalase(CAT) and malondialdehyde(MDA) in myocardial tissue. The levels of silent mating type information regulation 2homolog 1(Sirt1), peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α), B-cell lymphoma 2(Bcl-2), Bcl-2associated X protein(Bax) and cysteine-containing aspartate specific protease 3(Caspase-3) were detected by Western blotting. Results: DOX could significantly reduce the body weight of mice(P<0.05), serum LDH activity was significantly increased(P<0.05), antioxidant enzyme activity significantly decreased(P<0.05);Oral D-ribose significantly decreased LDH levels(P<0.05) and increased antioxidant enzyme activity(P<0.05), increased Sirt1, PGC-1α, Bcl-2 protein expression levels(P<0.05), decreased Caspase-3, Bax protein expression levels(P<0.05). Conclusion: D-ribose could alleviate the DOX-induced acute cardiotoxicity by activating Sirt1/PGC-1α pathway, inhibiting oxidative stress and apoptosis.