剑叶龙血树总黄酮对缺氧复氧处理后H9c2心肌细胞的保护作用
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摘要
目的探讨剑叶龙血树总黄酮(DCF)对缺氧复氧处理后H9c2心肌细胞的保护作用。方法实验分为正常对照组、模型对照组和DCF大、中、小剂量组(20,10,5μg·m L~(-1)),给药24 h后进行缺氧4 h复氧16 h处理,采用流式细胞术及Western blotting方法测定细胞凋亡及凋亡蛋白表达。结果 DCF(10,5μg·m L~(-1))能够显著降低H9c2心肌细胞凋亡率,并且能够提高PI3K、Akt蛋白表达,降低Bax、Cleaved Caspase-3及Cleaved Caspase-8蛋白表达。结论 DCF能够保护缺氧复氧处理后H9c2心肌细胞,其机制可能是通过PI3K及细胞凋亡通路发挥作用。
Objective To investigate the anti-apoptotic effect of Dracaena cochinchinensis flavonoids( DCF) on H9 c2 cardiacmyocytes in a hypoxia/reoxygenation injury model and its mechanism. Methods The H9 c2 cardiac myocytes were randomly divided into five groups: normal control group,model control group,DCF group( 20,10,5 μg·m L~(-1)). Each group received the corresponding treatment. Cell survival rate was tested by CCK-8 methods. Apoptotic rate was evaluated by propidium iodide( PI) staining and flow cytometry( FCM). The levels of PI3 K/Akt,Bax,Bcl-2,Cleaved Caspase-3,and cleaved Caspase-8 were evaluated by Western blotting. Results The DCF( 10,5 μg · m L~(-1)) could significantly protect the H9 c2 cardiacmyocytes from the hypoxia/reoxygenation injury with increasing cell survival rate. It also decreased the apoptotic percentage and unregulated the level of PI3 K/Akt and inhibited the expression of Bax,Cleaved Caspase-3,Cleaved Caspase-8.Conclusion DCF has the effect of anti-apoptosis on H9 c2 cardiacmyocytes exposed to hypoxia/reoxygenation injury via PI3 K/Akt and apoptotic signal pathway.
Objective To investigate the anti-apoptotic effect of Dracaena cochinchinensis flavonoids( DCF) on H9 c2 cardiacmyocytes in a hypoxia/reoxygenation injury model and its mechanism. Methods The H9 c2 cardiac myocytes were randomly divided into five groups: normal control group,model control group,DCF group( 20,10,5 μg·m L~(-1)). Each group received the corresponding treatment. Cell survival rate was tested by CCK-8 methods. Apoptotic rate was evaluated by propidium iodide( PI) staining and flow cytometry( FCM). The levels of PI3 K/Akt,Bax,Bcl-2,Cleaved Caspase-3,and cleaved Caspase-8 were evaluated by Western blotting. Results The DCF( 10,5 μg · m L~(-1)) could significantly protect the H9 c2 cardiacmyocytes from the hypoxia/reoxygenation injury with increasing cell survival rate. It also decreased the apoptotic percentage and unregulated the level of PI3 K/Akt and inhibited the expression of Bax,Cleaved Caspase-3,Cleaved Caspase-8.Conclusion DCF has the effect of anti-apoptosis on H9 c2 cardiacmyocytes exposed to hypoxia/reoxygenation injury via PI3 K/Akt and apoptotic signal pathway.
引文
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[2]钟月平.中药龙血竭的临床应用及研究进展[J].现代中西医结合杂志,2010,19(19):2469-2470.
[3]董恩双.龙血竭化学研究及降血糖有效部位的筛选[D].北京:北京中医药大学,2008:15-16.
[4] GUPTA D,BLEAKLEY B,GUPTA R K.Dragon's blood:botany,chemistry and therapeutic uses[J]. J Ethnopharm,2008,115(3):361-380.
[5]王兰珍,王继永,张燕,等.血竭植物资源开发利用现状[J].时珍国医国药,2007,18(4):1017-1019.
[6] WANG M,MENG X B,YU Y L,et al.Elatoside C protects against hypoxia/reoxygenation-induced apoptosis in H9c2cardiomyocytes through the reduction of endoplasmic reticulum stress partially depending on STAT3 activation[J].Apoptosis,2014,19(12):1727-1735.
[7] JEONG J J,HA Y M,JIN Y C,et al.Rutin from Lonicera japonica inhibits myocardial ischemia/reperfusion-induced apoptosis in vivo and protects H9c2 cells against hydrogen peroxide-mediated injury via ERK1/2 and PI3K/Akt signals in vitro[J].Food Chem Toxi,2009,47(7):1569-1576.
[8]李光,邢小燕,张美双,等.基于表观遗传学调控的中医药防治心肌缺血再灌注损伤的研究进展[J].药学学报,2016,51(7):1047-1053.
[9] ZHANG X,CHEN S,HU H,et al.Study on eqvivalence of dragon's blood and resina draconis on the cardiovascular system[J].China Pharmacist,2009,12(3):241-271.
[10]黄树芳,陈学芬,陈晓军,等.广西血竭的活血化瘀研究[J].中药材,1994,17(9):37-39,56.
[11]屠鹏飞,王钰芳,邵杰,等.龙血竭中黄酮类成分提取工艺研究[J].中国实验方剂学杂志,2011,17(6):30-32.
[12]秦建平,李家春,吴建雄,等.基于UPLC结合化学计量学方法的龙血竭指纹图谱研究[J].中国中药杂志,2015,40(6):1114-1118.
[13] KIMES B W,BRANDT B L.Properties of a clonal muscle cell line from rat heart.[J]. Exp Cell Res,1976,98(2):367-381.
[14]周艳林,蒋受军,文建文,等.龙血竭特征成分对照指纹图谱研究[J].中华中医药杂志,2012,27(12):3080-3082.
[15]刘伟,于沛,王玉强.白藜芦醇衍生物的药理活性研究进展[J].医药导报,2010,29(7):901-906.
[16]刘芝兰,曾春娇,吴梅青,等.白藜芦醇与白皮杉醇及赤松素研究进展[J].医药导报,2013,32(8):1043-1049.