蒙药额尔敦乌日勒预处理对心肌缺血再灌注损伤大鼠心肌组织自噬相关蛋白表达的影响
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摘要
目的探讨蒙药额尔敦-乌日勒预处理抗心肌缺血再灌注损伤的可能作用机制。方法将60只SD大鼠随机分为假手术组、模型组、复方丹参滴丸组和蒙药高、中、低剂量组,每组10只。蒙药高、中、低剂量组分别给予蒙药混悬液每次1. 2、0. 6、0. 3 g/kg灌胃,复方丹参滴丸组给予复方丹参滴丸溶液每次0. 5 g/kg灌胃,假手术组和模型组给予等体积蒸馏水,均每日2次,连续给药2周。末次给药12 h后,除假手术组只穿线不结扎外,其余各组均采用左冠状动脉前降支结扎法制备心肌缺血再灌注损伤模型。造模成功后HE染色观察各组大鼠心肌组织病理学变化,免疫组化检测心肌组织中自噬相关蛋白Beclin1和微管相关蛋白轻链3蛋白Ⅱ(LC3-Ⅱ)蛋白表达,Western blot法检测心肌组织中Beclin1、LC3-Ⅱ蛋白及PI3K-Aktm TOR信号通路相关蛋白的表达。结果与模型组比较,各给药组能够不同程度改善大鼠心肌组织的病理形态;蒙药中、高剂量组大鼠心肌组织中LC3-Ⅱ及Beclin1蛋白表达均较模型组下降(P <0. 05);蒙药中、高剂量组大鼠心肌组织中PI3K-Akt-mTOR信号通路相关蛋白相对表达量较模型组均升高(P <0. 05)。结论蒙药额尔敦-乌日勒可减轻心肌缺血再灌注损伤大鼠心肌组织损伤程度,可能与其下调自噬相关蛋白LC3-Ⅱ及Beclin1的表达,从而激活PI3K-Akt-mTOR信号通路有关。
Objective To explore the possible mechanism of anti-myocardial ischemia-reperfusion injury of Mongolian medicine E'erdun-Wurile pretreatment. Methods A total of 60 SD rats were randomly divided into sham operation group,model group,Compound Danshen Dropping Pills group and Mongolian medicine high,medium and low dose group,with 10 rats in each group. The Mongolian medicine high,medium,and low-dose groups were given the E'erdun-Wurile suspension respectively at a dose of 1. 2,0. 6,and 0. 3 g/kg each time,while Compound Danshen Dropping Pills group was given compound Danshen Dropping Pill solution at a dose of 0. 5 g/kg each time,and the sham operation group and the model group were given equal volume of distilled water twice daily for 2 weeks. Twelve hours after the last administration,the sham operation group was treated with only the thread not ligated,the other groups were treated with left anterior descending coronary artery ligation to prepare myocardial ischemia-reperfusion injury model. After successful modeling,the pathological changes of myocardial tissue in each group were observed by HE staining. The expressions of autophagy-related proteins Beclin1 and microtubule-associated protein light chian 3 protein Ⅱ( LC3-Ⅱ) were detected by immunohistochemistry. The Beclin1 and LC3-Ⅱ proteins in myocardial tissue and expression of PI3 K-Akt-mTOR signaling pathway-associated proteins were detected by Western blot. Results Compared with the model group,all medicated drug groups could improve the pathological morphology of rat myocardial tissue. The expressions of LC3-Ⅱ and Beclin1 in the myocardial tissues of the Mongolian medicine middle and high dose groups were lower than those in the model group( P < 0. 05),while the relative expression of PI3 K-Aktm TOR signaling pathway in the myocardial tissue was higher than that of the model group( P < 0. 05). Conclusion Mongolian medicine E'erdun-Wurile can alleviate myocardial tissue damage in rats with myocardial ischemia-reperfusion injury,which may be related to down-regulation of autophagy-related proteins LC3-Ⅱ and Beclin1 expression and activation of PI3 K-Akt-mTOR signaling pathway.
Objective To explore the possible mechanism of anti-myocardial ischemia-reperfusion injury of Mongolian medicine E'erdun-Wurile pretreatment. Methods A total of 60 SD rats were randomly divided into sham operation group,model group,Compound Danshen Dropping Pills group and Mongolian medicine high,medium and low dose group,with 10 rats in each group. The Mongolian medicine high,medium,and low-dose groups were given the E'erdun-Wurile suspension respectively at a dose of 1. 2,0. 6,and 0. 3 g/kg each time,while Compound Danshen Dropping Pills group was given compound Danshen Dropping Pill solution at a dose of 0. 5 g/kg each time,and the sham operation group and the model group were given equal volume of distilled water twice daily for 2 weeks. Twelve hours after the last administration,the sham operation group was treated with only the thread not ligated,the other groups were treated with left anterior descending coronary artery ligation to prepare myocardial ischemia-reperfusion injury model. After successful modeling,the pathological changes of myocardial tissue in each group were observed by HE staining. The expressions of autophagy-related proteins Beclin1 and microtubule-associated protein light chian 3 protein Ⅱ( LC3-Ⅱ) were detected by immunohistochemistry. The Beclin1 and LC3-Ⅱ proteins in myocardial tissue and expression of PI3 K-Akt-mTOR signaling pathway-associated proteins were detected by Western blot. Results Compared with the model group,all medicated drug groups could improve the pathological morphology of rat myocardial tissue. The expressions of LC3-Ⅱ and Beclin1 in the myocardial tissues of the Mongolian medicine middle and high dose groups were lower than those in the model group( P < 0. 05),while the relative expression of PI3 K-Aktm TOR signaling pathway in the myocardial tissue was higher than that of the model group( P < 0. 05). Conclusion Mongolian medicine E'erdun-Wurile can alleviate myocardial tissue damage in rats with myocardial ischemia-reperfusion injury,which may be related to down-regulation of autophagy-related proteins LC3-Ⅱ and Beclin1 expression and activation of PI3 K-Akt-mTOR signaling pathway.
引文
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[14]LI Z,DONG H,LI M,et al. Honokiol induces autophagy and apoptosis of osteosarcoma through PI3K/Akt/m TOR signaling pathway[J]. Mol Med Rep,2018,17(2):2719-2723.
[15]ZHANG J,WANG C,YU S,et al. Sevoflurane postconditioning protects rat hearts against ischemia-reperfusion injury via the activation of PI3K/AKT/m TOR signaling[J]. Sci Rep,2014,4(4):7317-7328. doi:10. 1038/srep07317.
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[18]KANG R,ZEH HJ,LOTZE MT,et al. The Beclin 1 network regulatesautophagy and apoptosis[J]. Cell Death Differ,2011,18(4):571-580.
[19]刘利娟,陈瑶,周德生,等. Salubrinal对大鼠脑缺血再灌注模型LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA的影响[J].广东医学,2017,38(4):509-513.
[20]刘宇,赵雅宁,刘孜卓,等. LC3-Ⅱ和Beclin1在糖尿病脑缺血后处理大鼠海马CA1区表达及意义[J].医学研究生学报,2016,29(1):46-51.
[2]MA S,WANG Y,CHEN Y,et al. The role of the autophagy in myocardial ischemia/reperfusion injury[J]. Biochim Biophys Acta,2015,1852(2):271-276.
[3]咏梅.蒙医心刺痛血性心刺痛的诊疗方案[J].中国民族医药杂志,2010,16(7):32-33.
[4]苏宝音,巴图那松.心刺痛的治疗[J].中国蒙医药,2013,8(4):33-35.
[5]松林.珍宝丸与沉香-35味散合用的苏荣扎布学术思想[J].中国蒙医药,2014,9(9):91-100.
[6]李明,鲁卫星,崔现军,等.提高大鼠心肌缺血/再灌注损伤模型成功率的方法探讨[J].辽宁中医杂志,2010,37(9):1699-1700.
[7]麻春杰,董平,韩雪梅,等.额尔敦-乌日勒在家兔动脉粥样硬化斑块消退中的作用及其对炎症因子的影响[J].世界科学技术—中医药现代化,2011,13(5):894-898.
[8]乌兰其其格,麻春杰,呼日乐巴根,等.额尔敦-乌日勒对兔动脉粥样化易损斑块模型血清超敏C反应蛋白及相关炎症因子的影响[J].环球中医药,2014,7(3):183-186.
[9]董平,麻春杰,韩雪梅,等.额尔敦-乌日勒对动脉粥样硬化家兔血清中NO和抗氧化酶活性的影响[J].中国医药,2011,6(11):1296-1298.
[10]董平,麻春杰,温都苏毕力格,等.额尔敦-乌日勒对实验性家兔动脉粥样硬化的影响[J].中华中医药学刊,2011,29(5):1012-1015.
[11]郭玉明,张弛,查青林,等.复方丹参滴丸治疗冠心病的系统评价[J].上海中医药大学学报,2012,26(3):24-31.
[12]LIU L,WU Y,HUANG X. Orientin protects myocardial cells against hypoxia-reoxygenation injury through induction of autophagy[J]. Eur J Pharmacol,2016,776:90-98.doi:10. 1016/j. ejphar. 2016. 02. 037.
[13]JIAN J,XUAN F,QIN F,et al. Bauhinia championⅡflavone inhibits apoptosis and autophagy via the PI3K/Akt pathway in myocardial ischemia/reperfusion injury in rats[J]. Drug Des Devel Ther,2015,9:5933-5945. doi:10.2147/DDDT. S92549.
[14]LI Z,DONG H,LI M,et al. Honokiol induces autophagy and apoptosis of osteosarcoma through PI3K/Akt/m TOR signaling pathway[J]. Mol Med Rep,2018,17(2):2719-2723.
[15]ZHANG J,WANG C,YU S,et al. Sevoflurane postconditioning protects rat hearts against ischemia-reperfusion injury via the activation of PI3K/AKT/m TOR signaling[J]. Sci Rep,2014,4(4):7317-7328. doi:10. 1038/srep07317.
[16]GALLO S,SALA V,GATTI S,et al. Cellular and molecular mechanisms of HGF/Met in the cardiovascular system[J]. Clin Sci(Lond),2015,129(12):1173-1193.
[17]CARROLL B,DUNLOP EA. The lysosome:a crucial hub for AMPKand m TORC1 signalling[J]. Biochem J,2017,474(9):1453-1466.
[18]KANG R,ZEH HJ,LOTZE MT,et al. The Beclin 1 network regulatesautophagy and apoptosis[J]. Cell Death Differ,2011,18(4):571-580.
[19]刘利娟,陈瑶,周德生,等. Salubrinal对大鼠脑缺血再灌注模型LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA的影响[J].广东医学,2017,38(4):509-513.
[20]刘宇,赵雅宁,刘孜卓,等. LC3-Ⅱ和Beclin1在糖尿病脑缺血后处理大鼠海马CA1区表达及意义[J].医学研究生学报,2016,29(1):46-51.