不同粒径天麻超微粉对脑缺血再灌注脑损伤大鼠的保护作用
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摘要
目的:研究不同粒径的天麻超微粉对局灶性脑缺血再灌注损伤大鼠的保护作用,为超微粉技术在天麻相关产品中的应用提供理论依据。方法:SD大鼠随机分为假手术组,模型组,天麻超微粉Ⅰ组(1. 2 g·kg-1),天麻超微粉Ⅱ组(1. 2 g·kg-1),天麻超微粉Ⅲ组(1. 2 g·kg-1),阳性药尼莫地平组(12. 6 g·kg-1),连续口服给药7 d后造模。采用单侧线栓法闭塞中动脉1 h后再灌注24 h,制备局灶性缺血再灌注损伤模型,2,3,5-氯化二苯基四氮唑(TTC)染色法测定脑组织梗死面积,比色法测定脑组织匀浆液和血清中的超氧化物歧化酶(SOD)的活性、丙二醛(MDA)含量、一氧化氮合酶(NOS)活性,液相色谱串联质谱法(LC/MSMS)记录脑海马中谷氨酸(Glu),天冬氨酸(Asp),甘氨酸(Gly)含量变化。结果:与假手术组比较,模型组出现显著的神经行为学功能缺失体征(P <0. 01),脑梗死面积显著增加(P <0. 01)。与模型组比较,天麻超微粉Ⅰ,Ⅱ,Ⅲ组均能明显改善神经行为学功能(P <0. 05),明显降低脑梗死面积比,Glu,Asp,MDA含量和NOS活性(P <0. 05,P <0. 01),明显升高SOD活性和Gly含量(P <0. 05)。与天麻超微粉Ⅰ组比较,天麻超微粉Ⅲ组SOD活性和Gly含量明显高于超微粉Ⅰ组(P <0. 05),Glu,Asp,MDA含量和NOS活性均明显低于超微粉Ⅰ组(P <0. 05),天麻超微粉Ⅱ组则均无显著性变化。结论:不同粒径的天麻超微粉均对脑缺血再灌注损伤具有预防保护作用,但天麻超微粉Ⅲ较天麻超微粉Ⅰ,Ⅱ预防保护作用较明显,表明天麻超微粉对大鼠大脑中动脉缺血再灌注的脑保护作用与其粒径大小具有一定相关性。
Objective: To study the protective effect of Gastrodiae Rhizoma ultrafine powder with different particle sizes on focal cerebral ischemia-reperfusion injury in rats,in order to provide theoretical basis for the application of ultrafine powder technology in related products. Method: The SD rats were randomly divided into six groups,namely sham-operated group,ischemia-reperfusion group,Gastrodiae Rhizoma ultramicro-powder Ⅰ group( 1. 2 g·kg-1),Gastrodiae Rhizoma ultramicro-powder Ⅱ group( 1. 2 g·kg-1),Gastrodiae Rhizoma ultramicropowder Ⅲ group( 1. 2 g·kg-1),and nimodipine positive drug group( 12. 6 g·kg-1). The cerebral ischemiareperfusion model was established after 7 days of drug administration. The focal ischemia reperfusion injury model was prepared by middle cerebral artery occlusion( MCAO) for 1 h followed by reperfusion for 24 h. The infarct size of brain tissue was measured by triphenyltetrazolium Chloride( TTC) staining. The activities of superoxide dismutase( SOD),malondialdehyde( MDA) and nitricoxide synthase( NOS) in brain homogenate and serum were measured by colorimetry. The contents of glutamate( Glu), aspartic acid( Asp) and glycine( Gly) in hippocampus were determined by liquid chromatography-tandem mass spectrometry( LC/MSMS). Result: The model group showed obvious signs of neurobehavioral deficit( P < 0. 01) compared with the sham operation group.The area ratio of cerebral infarction was decreased significantly,the contents of Glu,Asp,MDA and the activity of NOS were significantly decreased,while the activity of SOD and content of Gly were significantly increased( P <0. 05,P < 0. 01) in Gastrodiae Rhizoma ultramicro-powder Ⅰ,Ⅱ,Ⅲ groups compared with the model group.SOD activity and Gly content in the Gastrodiae Rhizoma ultramicro-powder Ⅲ group were significantly higher than those in the Gastrodiae Rhizoma ultramicro-powder Ⅰ group and the Gastrodiae Rhizoma ultramicro-powder Ⅱgroup( P < 0. 05),and Glu,Asp,MDA content and NOS activity in the Gastrodiae Rhizoma ultramicro-powder Ⅰgroup were significantly lower than those in the Gastrodiae Rhizoma ultramicro-powder Ⅱ group( P < 0. 05).Conclusion: Gastrodiae Rhizoma ultramicro-powder with different particle sizes has a protective effect on cerebral ischemia-reperfusion injury,and the Gastrodiae Rhizoma ultramicro-powder Ⅲ group has a more obvious protective effect than the the Gastrodiae Rhizoma ultramicro-powder Ⅰ group. The results showed that the protective effect of Gastrodiae Rhizoma ultramicro-powder on middle cerebral artery ischemia-reperfusion in rats was related to its particle size.
Objective: To study the protective effect of Gastrodiae Rhizoma ultrafine powder with different particle sizes on focal cerebral ischemia-reperfusion injury in rats,in order to provide theoretical basis for the application of ultrafine powder technology in related products. Method: The SD rats were randomly divided into six groups,namely sham-operated group,ischemia-reperfusion group,Gastrodiae Rhizoma ultramicro-powder Ⅰ group( 1. 2 g·kg-1),Gastrodiae Rhizoma ultramicro-powder Ⅱ group( 1. 2 g·kg-1),Gastrodiae Rhizoma ultramicropowder Ⅲ group( 1. 2 g·kg-1),and nimodipine positive drug group( 12. 6 g·kg-1). The cerebral ischemiareperfusion model was established after 7 days of drug administration. The focal ischemia reperfusion injury model was prepared by middle cerebral artery occlusion( MCAO) for 1 h followed by reperfusion for 24 h. The infarct size of brain tissue was measured by triphenyltetrazolium Chloride( TTC) staining. The activities of superoxide dismutase( SOD),malondialdehyde( MDA) and nitricoxide synthase( NOS) in brain homogenate and serum were measured by colorimetry. The contents of glutamate( Glu), aspartic acid( Asp) and glycine( Gly) in hippocampus were determined by liquid chromatography-tandem mass spectrometry( LC/MSMS). Result: The model group showed obvious signs of neurobehavioral deficit( P < 0. 01) compared with the sham operation group.The area ratio of cerebral infarction was decreased significantly,the contents of Glu,Asp,MDA and the activity of NOS were significantly decreased,while the activity of SOD and content of Gly were significantly increased( P <0. 05,P < 0. 01) in Gastrodiae Rhizoma ultramicro-powder Ⅰ,Ⅱ,Ⅲ groups compared with the model group.SOD activity and Gly content in the Gastrodiae Rhizoma ultramicro-powder Ⅲ group were significantly higher than those in the Gastrodiae Rhizoma ultramicro-powder Ⅰ group and the Gastrodiae Rhizoma ultramicro-powder Ⅱgroup( P < 0. 05),and Glu,Asp,MDA content and NOS activity in the Gastrodiae Rhizoma ultramicro-powder Ⅰgroup were significantly lower than those in the Gastrodiae Rhizoma ultramicro-powder Ⅱ group( P < 0. 05).Conclusion: Gastrodiae Rhizoma ultramicro-powder with different particle sizes has a protective effect on cerebral ischemia-reperfusion injury,and the Gastrodiae Rhizoma ultramicro-powder Ⅲ group has a more obvious protective effect than the the Gastrodiae Rhizoma ultramicro-powder Ⅰ group. The results showed that the protective effect of Gastrodiae Rhizoma ultramicro-powder on middle cerebral artery ischemia-reperfusion in rats was related to its particle size.
引文
[1]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,2018:58.
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[6]王正荣,罗红淋,肖静,等.天麻素对动脉血管顺应性以及血流动力学的影响[J].生物医学工程学杂志,1994,17(3):197-201.
[7]徐坚,陶陶,何燕,等.天麻及电针对大鼠脑缺血/再灌注损伤的保护作用[J].中国脑血管病杂志,2004,5(1):221-224.
[8]段小花,代蓉,李秀芳,等.天麻酚类成分对脑缺血模型大鼠海马一氧化氮和一氧化氮合酶的影响[J].中华老年心脑血管病杂志,2011,13(7):653-655.
[9]傅建华,杜贵友.天麻促智颗粒对反复脑缺血再灌注小鼠脑组织递质性氨基酸含量的影响[J].中国中药杂志,2001,26(1):54-56.
[10]李宏岳,范蕾,范蕾.沉香超微粉与山沉香超微粉药效学对比分析[J].中国实验方剂学杂志,2018,24(12):92-98.
[11]刘智,李诚秀,李玲,等.天麻粉不同粒径的镇静镇痛作用研究[J].中国现代应用药学,2002,19(5):383-385.
[12]刘智,王丽.天麻细胞级微粉碎和普通粉碎质量标准对比实验研究[J].中国现代应用药学,2002,19(6):474-476.
[13] Longa E Z,Weinstein P R,Carlson S,et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke,1989,20(1):84-91.
[14]陶陶,徐坚,董佑忠,等.天麻超微粉与普通粉对大鼠脑缺血再灌注损伤神经细胞凋亡的研究[J].中国医药学报,2004,19(7):409-411,388.
[15]陈燕,王有志,安春志,等. LC-ESI-MS/MS测定大鼠海马和纹状体中氨基酸类神经递质的含量[J].今日药学,2013,23(2):65-69.
[16]肖爱娇,陈日新,康明非,等.热敏灸对脑缺血再灌注损伤大鼠SOD、MDA的影响[J].天津医药,2014,42(1):51-53.
[17]王启海,梁枫,韩军.蜂胶总黄酮对脑缺血再灌注大鼠脑水肿和脑组织内NO及NOS含量的影响[J].包头医学院学报,2015,31(10):1-3.
[18]韩进,万海同,李金辉,等. 3'-甲氧基葛根素对缺血再灌注大鼠脑内氨基酸动态变化的实验研究[J].中国中药杂志,2012,37(7):1023-1027.
[19]张颖,杨坚,刘罡一,等.芍药苷草汤对脑缺血再灌注偏瘫大鼠痉挛状态和氨基酸水平的影响[J].上海中医药杂志,2015,49(5):85-89.
[20]陶陶,周建,徐坚,等.天麻对大鼠大脑皮质神经元细胞缺氧后超微结构及Bax和Bcl-2表达的影响[J].中华中医药杂志,2013,28(4):946-951.
[21]周建,陶陶,徐坚,等.天麻对缺氧引起大鼠大脑皮质神经元细胞凋亡的保护作用[J].中华中医药杂志,2013,28(3):640-644.
[22]陶陶,曾可斌,胡常林.天麻超微粉对大鼠脑缺血再灌注损伤的神经保护作用[J].中医杂志,2005,46(4):294-296.
[2]张大燕,文欢,王伟,等.天麻对帕金森小鼠神经元保护作用机制的转录组学分析[J].中国实验方剂学杂志,2018,24(16):73-81.
[3]杨飞,王信,马传江,等.天麻加工炮制、成分分析与体内代谢研究进展[J].中国中药杂志,2018,43(11):2207-2215.
[4]汤小虎,李波,胡璘媛,等.天麻在《太平惠民和剂局方》中的配伍应用[J].山西中医学院学报,2012,13(3):75-76.
[5]任世兰,于龙顺,赵国举,等.天麻对血管阻力和耐缺血缺氧能力的影响[J].中草药,1992,23(6):302-304,335.
[6]王正荣,罗红淋,肖静,等.天麻素对动脉血管顺应性以及血流动力学的影响[J].生物医学工程学杂志,1994,17(3):197-201.
[7]徐坚,陶陶,何燕,等.天麻及电针对大鼠脑缺血/再灌注损伤的保护作用[J].中国脑血管病杂志,2004,5(1):221-224.
[8]段小花,代蓉,李秀芳,等.天麻酚类成分对脑缺血模型大鼠海马一氧化氮和一氧化氮合酶的影响[J].中华老年心脑血管病杂志,2011,13(7):653-655.
[9]傅建华,杜贵友.天麻促智颗粒对反复脑缺血再灌注小鼠脑组织递质性氨基酸含量的影响[J].中国中药杂志,2001,26(1):54-56.
[10]李宏岳,范蕾,范蕾.沉香超微粉与山沉香超微粉药效学对比分析[J].中国实验方剂学杂志,2018,24(12):92-98.
[11]刘智,李诚秀,李玲,等.天麻粉不同粒径的镇静镇痛作用研究[J].中国现代应用药学,2002,19(5):383-385.
[12]刘智,王丽.天麻细胞级微粉碎和普通粉碎质量标准对比实验研究[J].中国现代应用药学,2002,19(6):474-476.
[13] Longa E Z,Weinstein P R,Carlson S,et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke,1989,20(1):84-91.
[14]陶陶,徐坚,董佑忠,等.天麻超微粉与普通粉对大鼠脑缺血再灌注损伤神经细胞凋亡的研究[J].中国医药学报,2004,19(7):409-411,388.
[15]陈燕,王有志,安春志,等. LC-ESI-MS/MS测定大鼠海马和纹状体中氨基酸类神经递质的含量[J].今日药学,2013,23(2):65-69.
[16]肖爱娇,陈日新,康明非,等.热敏灸对脑缺血再灌注损伤大鼠SOD、MDA的影响[J].天津医药,2014,42(1):51-53.
[17]王启海,梁枫,韩军.蜂胶总黄酮对脑缺血再灌注大鼠脑水肿和脑组织内NO及NOS含量的影响[J].包头医学院学报,2015,31(10):1-3.
[18]韩进,万海同,李金辉,等. 3'-甲氧基葛根素对缺血再灌注大鼠脑内氨基酸动态变化的实验研究[J].中国中药杂志,2012,37(7):1023-1027.
[19]张颖,杨坚,刘罡一,等.芍药苷草汤对脑缺血再灌注偏瘫大鼠痉挛状态和氨基酸水平的影响[J].上海中医药杂志,2015,49(5):85-89.
[20]陶陶,周建,徐坚,等.天麻对大鼠大脑皮质神经元细胞缺氧后超微结构及Bax和Bcl-2表达的影响[J].中华中医药杂志,2013,28(4):946-951.
[21]周建,陶陶,徐坚,等.天麻对缺氧引起大鼠大脑皮质神经元细胞凋亡的保护作用[J].中华中医药杂志,2013,28(3):640-644.
[22]陶陶,曾可斌,胡常林.天麻超微粉对大鼠脑缺血再灌注损伤的神经保护作用[J].中医杂志,2005,46(4):294-296.
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