摘要
以调整氧化应激状态、激活磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/内皮型一氧化氮合酶(eNOS)信号通路为切入点,探讨瓜蒌薤白半夏汤(GXBD)保护Ⅱ型糖尿病合并急性心肌缺血(T2DM-AMI)的作用机制.将36只大鼠随机分为对照组(Ctrl组)、模型组(T2DM-AMI组)、处理组(GXBD组),每组12只.对T2DM-AMI组、GXBD组联合使用灌服高脂乳剂、腹腔注射链脲佐菌素、冠脉结扎3种方法制造T2DM-AMI大鼠模型.四唑红染色检测心肌梗死率,苏木精-伊红染色检测其病理损伤情况;酶联免疫吸附法检测血浆氧化损伤成分氧化型低密度脂蛋白(ox-LDL)、晚期糖基化终产物(AGEs)、黄嘌呤氧化酶(XO)、髓过氧化物酶(MPO),血浆抗氧化成分一氧化氮(NO)、硫氧还蛋白(Trx)、硫氧还蛋白还原酶(TrxR)的水平或活性,以及总活性氧(T-ROS)的水平和总抗氧化能力(T-AOC);蛋白免疫印迹法检测心肌组织中PI3K、Akt和eNOS蛋白的表达及磷酸化水平,实时荧光定量PCR检测其中Pi3k、Akt和eNos基因的mRNA表达水平.结果显示T2DM-AMI大鼠心肌梗死严重,血浆中上述4种氧化损伤成分的水平或活性及T-ROS水平显著升高,3种抗氧化成分的水平或活性及T-AOC显著降低,心肌组织中PI3K、Akt和eNOS蛋白的表达及磷酸化水平显著下调,相应基因的mRNA表达水平同样显著下降,而GXBD干预可显著逆转这些病理性变化.综上,GXBD可能通过提高抗氧化能力、清除自由基、纠正异常的氧化应激状态、激活心肌组织中PI3K/Akt/eNOS信号通路,发挥对T2DM-AMI的保护作用.
By adjusting oxidative stress state and activating PI3K/Akt/eNOS signaling pathway,this study explores the mechanism of Gualou Xiebai Banxia decoction(GXBD)in protecting typeⅡdiabetes mellitus complicated with acute myocardial ischemia(T2DM-AMI).36 rats were randomly divided to control(Ctrl)group,model(T2DM-AMI)group and treatment(GXBD)group,12 rats each.T2DM-AMI group and GXBD group were gavaged by high-fat emulsion,intrapitoneal injection of streptozotocin and coronary artery ligation.Tetrazolium Red(TTC)staining was used to detect the myocardial infarction rate,hematoxylin-eosin(HE)staining was used to detect the pathological damage.Enzyme-linked immuno sorbent assay was used to detect the levels or activities of oxidative stress components(ox-LDL,AGEs,XO,MPO)and anti-oxidative components(NO,Trx,TrxR)in plasma,and the total reactive oxygen species(T-ROS)level and total antioxidant capacity(T-AOC),and Western blotting was used to detect the protein expression and phosphorylation levels of PI3K,Akt and eNOS.The mRNA expression levels of Pi3k,Akt and eNos genes were detected by quantitative real-time PCR.Results showed that the T2DM-AMI rats had severe myocardial infarction,levels or activities of the four oxidative stress components and T-ROS level in plasma were significantly increased,levels or activities of the three anti-oxidative components and T-AOC were significantly decreased,expression and phosphorylation levels of the above three proteins in myocardial tissue were significantly down-regulated.mRNA expression levels of the three genes were also significantly reduced.However,GXBD intervention can significantly reverse these pathological changes.As a conclusion,GXBD plays a protective role against T2DM-AMI by improving anti-oxidative ability,removing radicals,rectifying abnormal oxidative stress state,activating PI3K/Akt/eNOS signaling pathway in myocardial tissue.
作者
孙笛洋
荀丽英
周继栋
杨玉婕
王蕾
赵启韬
SUN Diyang;XUN Liying;ZHOU Jidong;YANG Yujie;WANG Lei;ZHAO Qitao(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;College of Traditional Chinese Medicine,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2023年第1期119-126,共8页
Journal of Xiamen University：Natural Science
基金
国家自然科学基金(81573852)
山东省自然科学基金(ZR2019BH017)。
