摘要
目的:探究芝麻素(SES)对链脲佐菌素(STZ)诱导的糖尿病心肌病的影响及其机制。方法:将8周雄性C57BL/6J小鼠随机分为5组(每组8只):对照组、SES组、糖尿病心肌病(DCM)组、DCM+二甲双胍(Metformin,MET)灌胃组(简称DCM+MET组),以及DCM+SES灌胃组(简称DCM+SES组)。通过STZ腹腔注射的方法诱导DCM模型。造模成功12周后,给药SES和MET,4周后取出小鼠心脏用于病理和分子生物学分析。二氢乙啶(DHE)染色检测心肌组织活性氧水平,免疫组化染色检测心肌组织CD45和肿瘤坏死因子-α(TNF-α)表达水平;实时聚合酶链反应(RT-PCR)和免疫印迹检测相关分子表达变化。细胞培养Sirt3抑制试验后,将细胞分为8组:对照组[5.5 mmol/L葡萄糖(GLU)培养24 h]、SES组(10 mmol/L处理24 h)、高糖(HG)组(33 mmol/L GLU刺激24 h)、HG+SES组(33 mmol/L GLU+10 mmol/L SES 24 h)及在前四组的基础上分别添加Sirt3选择性抑制剂(3-TYP,50μmol/L处理24 h)即:对照+3-TYP组、SES+3-TYP组、HG+3-TYP组、HG+SES+3-TYP组。细胞实验验证SES对高糖刺激的细胞的具体机制。结果:本研究成功制备1型糖尿病心肌病模型,DCM+SES组小鼠餐后血糖水平显著低于DCM组(P<0.05)。DCM+SES组DHE阳性强度显著低于DCM组(P<0.05),DCM+SES组心肌组织丙二醛显著低于DCM组,而超氧化物歧化酶含量显著高于DCM组(P均<0.001);DCM+SES组小鼠的CD45和TNF-α阳性区域显著低于DCM组(P<0.05);DCM+SES组小鼠的Sirt3蛋白表达水平显著高DCM组(P<0.05)。H9c2细胞实验中,3-TYP的使用阻遏了SES的保护作用。HG+SES+3-TYP组与HG+3-TYP组细胞存活率差异均无统计学意义[(55.39±8.32)%vs.(59.67±7.42)%,P>0.05]。结论:SES可对STZ诱导的DCM具有显著的保护作用,其机制可能与降血糖、抑制氧化应激和炎症反应、上调Sirt3的表达有关。
Objectives:To investigate the effects and mechanisms of sesamin(SES)on diabetic cardiomyopathy induced by streptozotocin(STZ)in mice.Methods:Eight-week-old male C57BL/6J mice were randomly divided into 5 groups(n=8 in each group):control group,sesamin group,diabetic cardiomyopathy(DCM)group,DCM+metformin(MET)intragastric administration group and DCM+SES intragastric administration group.DCM model was induced by intraperitoneal injection of STZ.At 12 weeks post STZ,SES and MET were administered.After 4 weeks,the mouse hearts of each group were removed for pathological and molecular biology analysis.DHE staining was used to detect the level of reactive oxygen species in myocardium,immunohistochemical staining was used to detect the expression of CD45 and tumor necrosis factor-α(TNF-α),real-time polymerase-linked reaction(RT-PCR)and Western blotting were used to detect changes in the expression of related molecules The specific mechanism of sesamin was explored in vitro on cells stimulated by high glucose(HG).Results:In this study,the model of type 1 diabetic cardiomyopathy was successfully established.The level of blood glucose was significantly lower in DCM+SES group than that in DCM group(P<0.05).The positive intensity of DHE was significantly lower in DCM+SES group than that in DCM group(P<0.05),and the content of MDA/SOD in myocardium was significantly lower/higher in DCM+SES group than that in DCM group(P<0.001).The positive regions of CD45 and TNF-αwere significantly lower in DCM+SES group than those in DCM group(P<0.05).The expression level of Sirt3 protein was significantly higher in DCM+SES group than that in DCM group(P<0.05).In H9c2 cell experiment,the protective effect of sesamin was blocked by Sirt3 selective inhibitor(3-TYP).There was no significant difference in the cell viability between HG+SES+3-TYP group and HG+3-TYP group([55.39±8.32]%vs.[59.67±7.42]%,P>0.05).Conclusions:Sesamin could attenuate myocardial injury in STZ-induced diabetic cardiomyopathy mice model,and its mechanism may be related to lowering blood glucose,inhibiting oxidative stress and inflammation,and up-regulating the expression of Sirt3.
作者
郭振
刘方圆
安鹏
汪铭煜
杨丹
杨政
樊迪
唐其柱
GUO Zhen;LIU Fangyuan;AN Peng;WANG Mingyu;YANG Dan;YANG Zheng;FAN Di;TANG Qizhu(Department of Cardiology,Renmin Hospital of Wuhan University,Hubei Key Laboratory of Metabolic and Chronic Diseases,Wuhan(430060),Hubei,China)
出处
《中国循环杂志》
CSCD
北大核心
2021年第9期903-910,共8页
Chinese Circulation Journal
基金
国家自然科学基金(81900219,81800216)
中央高校基础研究经费(2042019kf0062)
国家重点研发计划(2018YFC1311300)
武汉市科技计划项目(2018061005132295)。
