加味丹参饮预处理调节ODC活性抗大鼠心肌缺血再灌注损伤研究
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摘要
目的探讨加味丹参饮通过调节鸟氨酸脱羧酶(ODC)活性在抗SD大鼠心肌缺血再灌注损伤(IRI)中的保护机制。方法建立SD大鼠心肌IRI模型,设对照组、假手术组、缺血再灌注损伤(IRI)组、加味丹参饮(JWDSY)+IRI组共4组。酶联免疫(ELISA)法测定ODC含量,实时荧光定量PCR(RT-PCR)方法检测ODC m RNA表达水平,免疫印迹(Western blot)法检测ODC蛋白表达水平,高效液相色谱检测心肌组织中多胺的含量。结果与对照组比较,假手术组的ODC含量、ODC m RNA及蛋白表达水平均无统计学意义(P>0.05);与假手术组比较,IRI组的ODC含量显著增加、ODC m RNA及蛋白表达水平显著升高,差异有统计学意义(P<0.01);与IRI组比较,JWDSY+IRI组ODC含量显著减少,ODC m RNA和蛋白表达水平显著降低,差异有统计学意义(P<0.01)。结论加味丹参饮可能通过调节ODC活性,从而发挥对IRI后心肌的保护作用。
Objective To explore the protective mechanism of modified Danshen decoction on SD rats myocardial ischemia-reperfusion injury(IRI) by ornithine decarboxylase(ODC) activity. Methods The SD rat models of myocardial IRI were established, the four groups of control group, sham-operation group, ischemia/reperfusion(IRI) group, Jiawei Danshen Decoction(JWDSY) + IRI group were assigned. The content of ODC was determined by ELISA method. The ODC m RNA expression level was detected by fluorescence quantitative RT-PCR method. The ODC protein expression level was detected by Western blot method, and the polyamine content was determined by HPLC. Results Compared with the control group, the content of ODC, ODC m RNA and protein expression level in sham-operationa group were increased, the difference had no statistically significant(P>0.05), while compared with the sham-operation group, the content of ODC, ODC m RNA and protein expression level in IRI group were increased significantlay(P<0.01). Compared with IRI group, the ODC content in JWDSY + IRI group was significantly reduced, and the ODC m RNA and protein expression level were decreased significantly, the difference was statistically significant(P<0.01). Conclusion Modified Danshen decoction could adjust the ODC activity, and then play a protective effect on myocardium after IRI.
Objective To explore the protective mechanism of modified Danshen decoction on SD rats myocardial ischemia-reperfusion injury(IRI) by ornithine decarboxylase(ODC) activity. Methods The SD rat models of myocardial IRI were established, the four groups of control group, sham-operation group, ischemia/reperfusion(IRI) group, Jiawei Danshen Decoction(JWDSY) + IRI group were assigned. The content of ODC was determined by ELISA method. The ODC m RNA expression level was detected by fluorescence quantitative RT-PCR method. The ODC protein expression level was detected by Western blot method, and the polyamine content was determined by HPLC. Results Compared with the control group, the content of ODC, ODC m RNA and protein expression level in sham-operationa group were increased, the difference had no statistically significant(P>0.05), while compared with the sham-operation group, the content of ODC, ODC m RNA and protein expression level in IRI group were increased significantlay(P<0.01). Compared with IRI group, the ODC content in JWDSY + IRI group was significantly reduced, and the ODC m RNA and protein expression level were decreased significantly, the difference was statistically significant(P<0.01). Conclusion Modified Danshen decoction could adjust the ODC activity, and then play a protective effect on myocardium after IRI.
引文
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[11]赵雅君,王丽娜,李鸿珠,等.大鼠心肌多胺代谢限速酶ODC、SSAT活性分析[J].中国药理学通报,2005,21(5):555-559.
[12]万新,姜冰.多胺与脑缺血[J].国际神经病学神经外科学杂志,2001,28(6):453-455.
[13]Zhao YJ,Zhang WH,Xu CQ,et al.Involvement of the ornithine decarboxylase/polyamine system in precondition-induced cardioprotection through an interaction with PKC in rat hearts[J].Molecular and cellular Biochemistry,2009,3271/2:135-144.
[14]张国中,刘增长.内质网应激与心肌缺血再灌注损伤[J].实用临床医药杂志,2015,19(7):4714-4717.
[2]彭霞,黄政德,成细华,等.加味丹参饮预处理调控自噬相关基因Beclin-1和Atg5表达抗大鼠心肌缺血再灌注损伤[J].湖南中医药大学学报,2016,36(7):20-24.
[3]陈聪,任婷,胡华,等.加味丹参饮预处理对大鼠心肌缺血再灌注损伤的保护作用[J].湖南中医药大学学报,2016,36(6):11-15.
[4]任婷,黄政德,田雪飞,等.心痛舒含药血清预处理对乳鼠心肌细胞缺氧/复氧损伤及TNF-α、IL-1β的影响[J].湖南中医药大学学报,2012,32(11):11-15.
[5]Livak KJ,Schmittgen TD.Analysis of relative gene expression data using real-time quantitative PCR and the 2-荭荭GMethod[J].Methods,2001,25(4):402-408.
[6]赵雅君,徐长庆,李全风,等.应用反向高效液相色谱分析离体心脏灌流大鼠心肌组织中的多胺[J].中国病理生理杂志,2005,21(2):412-413.
[7]Nadège Minois,Didac Carmona-Gutierrez,Frank Madeo.Polyamines in aging and disease[J].AGING,2011,3(8):716-732.
[8]Jnne J,Alhonen L,PietilM,et al.Genetic approaches to the cellular functions of polyamines in mammals[J].European journal of biochemistry,2004,271(5):877-894.
[9]Puntambekar SS,Davis DS,Hawel III L,et al.LPS-induced CCL2 expression and macrophage influx into the murine central nervous system is polyamine-dependent[J].Brain,Behavior,and Immunity,2011,25(4):629-639.
[10]Dong H,Wang S,Zhang Z,et al.The effect of mitochondrial calcium uniporter opener spermine on diazoxide against focal cerebral ischemia-reperfusion injury in rats[J].Stroke Cerebrovasc Dis,2014,23(2):303-309.
[11]赵雅君,王丽娜,李鸿珠,等.大鼠心肌多胺代谢限速酶ODC、SSAT活性分析[J].中国药理学通报,2005,21(5):555-559.
[12]万新,姜冰.多胺与脑缺血[J].国际神经病学神经外科学杂志,2001,28(6):453-455.
[13]Zhao YJ,Zhang WH,Xu CQ,et al.Involvement of the ornithine decarboxylase/polyamine system in precondition-induced cardioprotection through an interaction with PKC in rat hearts[J].Molecular and cellular Biochemistry,2009,3271/2:135-144.
[14]张国中,刘增长.内质网应激与心肌缺血再灌注损伤[J].实用临床医药杂志,2015,19(7):4714-4717.