四逆汤及其拆方水煎液对缺血心肌细胞的保护作用
|
摘要
目的:探讨四逆汤(Sini decoction,SND)及其拆方水煎液对缺血大鼠离体心肌细胞H_9C_2的保护作用。方法:采用不同浓度的含药培养液培养细胞,显微镜下观察心肌细胞建模前后的生长状态;利用MTT比色法比较各加药浓度下单味药、药对、全方对缺血前后心肌细胞H_9C_2存活率的影响,并分析各试药之间作用的差异。结果:1)400倍镜下可见正常培养细胞生长状态良好,细胞形态为梭形,聚集式生长;缺血模型细胞形态改变,间隙变大,结构发生萎缩,死亡细胞增多。2)与缺血后SND组比较,10、20、5 mg/mL浓度干姜水煎液(Ginger water decoction,GWD)对缺血后H_9C_2细胞无保护作用,存活率分别为(44.89±2.59)%、(49.43±1.87)%、(73.67±1.30)%,两两之间差异显著(P<0.01)。在浓度为1.0 mg/mL时,附子水煎液(aconite water decoction,AWD)对缺血前后各组细胞均产生细胞毒性。甘草水煎液(Licorice water decoction,LWD)同附子-甘草水煎液(licorice and aconite decoction,LAWD)比较,对H_9C_2保护作用均较弱。附子-干姜水煎液(Ginger and aconite water decoction,GAWD)各浓度缺血后加药组同SND组比较对H_9C_2保护作用无差异。结论:SND及其拆方GAWD、LAWD、AWD、LWD均对缺血H_9C_2有明显促增殖作用;各浓度GWD对缺血H_9C_2无保护作用,1.0 mg/mL AWD有细胞毒性。
Objective: To study the protective effects of SiNi decoction(SND) and the decoction of the constituent drugs on isolated ischemic myocardial cells H_9C_2 of the rats. Methods: Different concentrations of medicated nutrient fluids were used to culture the cells, the growth conditions of myocardial cells were observed before and after establishing the models under the microscope; MTT chromatometry was used to compare the effects of single herb, herbal couplets and the whole prescriptions on survival rate of H_9C_2 before and after the ischemia, and analyze the difference among the test herbs. Results: 1) Under 400 times microscope, it could be seen that the growth of normal cells was good, cellular morphology was fusiformis, growing in clusters; cellular morphology of the ischemic cells present changes, the gap became large, the structure atrophied, and the dead cells increased. 2) Compared with SND group after ischemia, 10,20 and 5 mg/mL ginger water decoction(GWD) showed no protective effects on H_9C_2 cells after the ischemia, the survival rates were(44.89 ±2.59)%,(49.43 ±1.87)% and(73.67 ±1.30)%respectively, the difference was significant between both(P<0.01). FuZi decoction(aconite water decoction, AWG)could produce cytotoxicity on the cells in different groups before and after ischemia when the decoction was in the concentrations of 1.0 mg/mL. When licorice water decoction(LWD) was compared with licorice and aconite decoction(LAWD), it showed weaker protective effects on H_9C_2. There was no difference when the medication groups of ginger and aconite water decoction(GAWD) in different concentrations after the ischemia was compared with SND group.Conclusion: SND and the decoction of the constituent drug pairs GAWD, LAWD, AWD and LWD could promote the proliferation of ischemic myocardial cells H_9C_2, different concentrations of GWD could not protect ischemic H_9C_2, while 1.0 mg/mL AWD shows cytotoxicity.
Objective: To study the protective effects of SiNi decoction(SND) and the decoction of the constituent drugs on isolated ischemic myocardial cells H_9C_2 of the rats. Methods: Different concentrations of medicated nutrient fluids were used to culture the cells, the growth conditions of myocardial cells were observed before and after establishing the models under the microscope; MTT chromatometry was used to compare the effects of single herb, herbal couplets and the whole prescriptions on survival rate of H_9C_2 before and after the ischemia, and analyze the difference among the test herbs. Results: 1) Under 400 times microscope, it could be seen that the growth of normal cells was good, cellular morphology was fusiformis, growing in clusters; cellular morphology of the ischemic cells present changes, the gap became large, the structure atrophied, and the dead cells increased. 2) Compared with SND group after ischemia, 10,20 and 5 mg/mL ginger water decoction(GWD) showed no protective effects on H_9C_2 cells after the ischemia, the survival rates were(44.89 ±2.59)%,(49.43 ±1.87)% and(73.67 ±1.30)%respectively, the difference was significant between both(P<0.01). FuZi decoction(aconite water decoction, AWG)could produce cytotoxicity on the cells in different groups before and after ischemia when the decoction was in the concentrations of 1.0 mg/mL. When licorice water decoction(LWD) was compared with licorice and aconite decoction(LAWD), it showed weaker protective effects on H_9C_2. There was no difference when the medication groups of ginger and aconite water decoction(GAWD) in different concentrations after the ischemia was compared with SND group.Conclusion: SND and the decoction of the constituent drug pairs GAWD, LAWD, AWD and LWD could promote the proliferation of ischemic myocardial cells H_9C_2, different concentrations of GWD could not protect ischemic H_9C_2, while 1.0 mg/mL AWD shows cytotoxicity.
引文
[1]窦有业,杜蓉.四逆汤的临床应用与实验研究进展[J].医药导报,2008,27(1):74-76.
[2]刘平,葛迎春,马天舒.四逆汤类方药理研究进展[J].辽宁中医杂志,2007,34(2):248-251.
[3]李莹.基于生物药剂学研究四逆汤中附子与甘草合煎减毒增效机制[D].成都:成都中医药大学,2013.
[4]Zhang H,Liu M,Zhang W,et al.Comparative pharmacokinetics of three monoester-diterpenoid alkaloids after oral administration of acontium carmichaeli extract and its compatibility with other herbal medicines in Sini decoction to rats[J].Biomedical Chromatography,2015,29(7):1083.
[5]Chen QQ,Han X,Wang WM,et al.Danggui Sini decoction ameliorates myelosuppression in animal model by upregulating thrombopoietin expression[J].Cell Biochemistry and Biophysics,2015,71(2):945-950.
[6]孙慧兰,吴伟康.四逆汤有效成分不同组合抗心肌缺血再灌注损伤的作用研究[J].中草药,2002,33(4):333-336.
[7]蔺建军.四逆汤干预大鼠急性缺血性心肌梗死模型的动态药效研究[D].济南:山东中医药大学,2012.
[8]李晓梅,包馨慧,杨毅宁,等.体外心肌细胞缺血后适应模型的建立及评价[J].新疆医科大学学报,2014,37(1):24-28.
[9]赵帅,李庆玲,韩旭,等.离体心肌细胞损伤模型研究进展[J].长春中医药大学学报,2015,31(3):650-653.
[10]张娜娟,张彤彤,刘志祥,等.体外心肌细胞缺血再灌注损伤模型的建立[J].四川动物,2009,28(3):440-443.
[11]国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:789.
[12]王达理,曾爱萍,南柏松.心肌细胞培养方法和活细胞噻唑兰法测定[J].空军医高专学报,1999,21(4):191-193.
[13]蒋燕萍,曹丽梅,彭伟,等.四逆汤及不同组方配伍对大鼠心肌缺血影响的对比研究[J].中草药,2016,47(4):630-633.
[14]贺金,方艳伟,李永民.四逆汤对大鼠心肌缺血损伤的保护作用[J].中华中医药杂志,2008,23(7):638-640.
[15]林董,何爱明,吴丽萍,等.MTT法测定8种中草药体外抗肝癌细胞SMMC-7721活性[J].安徽农业科学,2009,37(34):17264-17266.
[16]廖晖.四逆汤中甘草作用分析[J].时珍国医国药,2001,12(3):271-272.
[17]彭伟.四逆汤中药味配伍前后药效成分变化规律及药效差异研究[D].成都:成都中医药大学,2015.
[18]葛迎春,马天舒,任慧君,等.四逆汤类方不同工艺及配伍提取物对小鼠脾细胞增殖的影响[J].辽宁中医杂志,2010,37(8):1597-1599.
[2]刘平,葛迎春,马天舒.四逆汤类方药理研究进展[J].辽宁中医杂志,2007,34(2):248-251.
[3]李莹.基于生物药剂学研究四逆汤中附子与甘草合煎减毒增效机制[D].成都:成都中医药大学,2013.
[4]Zhang H,Liu M,Zhang W,et al.Comparative pharmacokinetics of three monoester-diterpenoid alkaloids after oral administration of acontium carmichaeli extract and its compatibility with other herbal medicines in Sini decoction to rats[J].Biomedical Chromatography,2015,29(7):1083.
[5]Chen QQ,Han X,Wang WM,et al.Danggui Sini decoction ameliorates myelosuppression in animal model by upregulating thrombopoietin expression[J].Cell Biochemistry and Biophysics,2015,71(2):945-950.
[6]孙慧兰,吴伟康.四逆汤有效成分不同组合抗心肌缺血再灌注损伤的作用研究[J].中草药,2002,33(4):333-336.
[7]蔺建军.四逆汤干预大鼠急性缺血性心肌梗死模型的动态药效研究[D].济南:山东中医药大学,2012.
[8]李晓梅,包馨慧,杨毅宁,等.体外心肌细胞缺血后适应模型的建立及评价[J].新疆医科大学学报,2014,37(1):24-28.
[9]赵帅,李庆玲,韩旭,等.离体心肌细胞损伤模型研究进展[J].长春中医药大学学报,2015,31(3):650-653.
[10]张娜娟,张彤彤,刘志祥,等.体外心肌细胞缺血再灌注损伤模型的建立[J].四川动物,2009,28(3):440-443.
[11]国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:789.
[12]王达理,曾爱萍,南柏松.心肌细胞培养方法和活细胞噻唑兰法测定[J].空军医高专学报,1999,21(4):191-193.
[13]蒋燕萍,曹丽梅,彭伟,等.四逆汤及不同组方配伍对大鼠心肌缺血影响的对比研究[J].中草药,2016,47(4):630-633.
[14]贺金,方艳伟,李永民.四逆汤对大鼠心肌缺血损伤的保护作用[J].中华中医药杂志,2008,23(7):638-640.
[15]林董,何爱明,吴丽萍,等.MTT法测定8种中草药体外抗肝癌细胞SMMC-7721活性[J].安徽农业科学,2009,37(34):17264-17266.
[16]廖晖.四逆汤中甘草作用分析[J].时珍国医国药,2001,12(3):271-272.
[17]彭伟.四逆汤中药味配伍前后药效成分变化规律及药效差异研究[D].成都:成都中医药大学,2015.
[18]葛迎春,马天舒,任慧君,等.四逆汤类方不同工艺及配伍提取物对小鼠脾细胞增殖的影响[J].辽宁中医杂志,2010,37(8):1597-1599.