玛咖性温健脾及对脾虚小鼠线粒体能量代谢酶的影响
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摘要
目的考察玛咖缓解脾虚证小鼠体力疲劳、抗氧化损伤作用,验证玛咖性温健脾中药药性,探讨能量代谢机制。方法雄性昆明种小鼠随机分为正常组,模型组,人参组,玛咖高、中、低剂量组,除正常组外其余各组均采用饮食失节、劳倦伤脾双因素法建立脾虚证小鼠模型,共计14 d,同时从第1天开始各组预防给药。记录脾虚小鼠力竭游泳时间,比色法测肝脏组织匀浆中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)活性或浓度。放免法测血清环磷酸腺苷(c AMP)和环磷酸鸟苷(c GMP)浓度,并计算比值(c AMP/c GMP)。连续循环法测肝脏组织线粒体F0F1-ATP酶活性,酶解法测Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性。结果与正常组比较,模型组小鼠体重下降,力竭游泳时间减少,肝组织中SOD、CAT、GSH-Px活力下降,MDA升高,c AMP/c GMP比值下降,F0F1-ATP酶、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性均降低(P<0.01)。与模型组比较,玛咖高、中、低剂量组(1.5、0.75、0.25 g/kg)均能增加脾虚小鼠体重,具有统计意义(P<0.01);玛咖高剂量组和人参组能显著延长脾虚小鼠力竭游泳时间(P<0.01);玛咖中、高剂量组同人参组均能显著提高脾虚小鼠肝脏组织中SOD、CAT、GSH-Px活力(P<0.01),减少MDA含量(P<0.01),显著升高血清c AMP/c GMP比值(P<0.01),显著增加F0F1-ATP酶、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性(P<0.01)。结论一定剂量的玛咖能够缓解脾虚小鼠体力疲劳,提高抗氧化酶的活性并消除自由基细胞毒性,验证了玛咖性温健脾的中药药性,进一步证实其机制与增强线粒体能量代谢酶相关。
Objective To investigate the effects of relieving physical fatigue and anti-oxidative damage of Maka(Maca,Lepidium meyenii Walp.),to verify Maka's properties of warming and fortifying spleen,and to discuss the mechanism of energy metabolism in mice with spleen deficiency pattern. Methods Male Kunming mice were randomly divided into normal group,model group,ginseng group(control group) and high-dose,mid-dose and low-dose Maka groups(1. 5 g/kg,0. 75 g/kg,0. 25 g/kg).Except of normal group,other groups were given abnormal diet and fatigue to impair spleen for 14 d for establishing mouse model of spleen deficiency pattern. Meanwhile all groups were preventively given corresponding drugs at the first day. The time of forced swimming test(FST) was recorded in spleen deficiency mice. The activities or concentration of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidease(GSH-PX) and malondialdehyde(MDA) in liver tissue homogenate were detected by using chromatoptometry. The concentration of serum c AMP and c GMP were detected by using radioimmunoassay(RIA),and ratio of c AMP to c GMP(c AMP/c GMP) was calculated. The activity of F0 F1-ATP enzyme was detected by using continuous cycle method,and activities of Na+-K+-ATP and Ca2 +-Mg2 +-ATP enzymes were detected by using enzymolysis. Results Compared with normal group,mouse body weight and time of FST decreased,activities of SOD,CAT and GSH-PX decreased,MDA content increased,c AMP/c GMP decreased,and activities of F0 F1-ATP,Na+-K+-ATP and Ca2 +-Mg2 +-ATP decreased in model group(P < 0. 01). Compared with model group,mouse body weight was increased in all Maka groups(P < 0. 01). The time of FST was significantly prolonged in high-dose Maka group and ginseng group(P < 0. 01),which was not significant in mid-dose and low-dose Maka groups. The activities of SOD,CAT and GSH-PX were significantly increased(P < 0. 01),MDA content was decreased(P < 0. 01),c AMP/c GMP was significantly increased(P < 0. 01),and activities of F0 F1-ATP,Na+-K+-ATP and Ca2 +-Mg2 +-ATP were significantly increased(P < 0. 01) in mid-dose and high-dose Maka groups,which was no significant in low-dose Maka group. Conclusion Maka,in a certain dose,can relieve physical fatigue,improve activities of anti-oxidative enzymes and eliminate cytotoxicity of free radicals,which verifies Maka's TCM properties of warming and fortifying spleen. The correlation between Maka's effective mechanism and enhancing mitochondrial energy metabolism enzymes is further confirmed.
Objective To investigate the effects of relieving physical fatigue and anti-oxidative damage of Maka(Maca,Lepidium meyenii Walp.),to verify Maka's properties of warming and fortifying spleen,and to discuss the mechanism of energy metabolism in mice with spleen deficiency pattern. Methods Male Kunming mice were randomly divided into normal group,model group,ginseng group(control group) and high-dose,mid-dose and low-dose Maka groups(1. 5 g/kg,0. 75 g/kg,0. 25 g/kg).Except of normal group,other groups were given abnormal diet and fatigue to impair spleen for 14 d for establishing mouse model of spleen deficiency pattern. Meanwhile all groups were preventively given corresponding drugs at the first day. The time of forced swimming test(FST) was recorded in spleen deficiency mice. The activities or concentration of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidease(GSH-PX) and malondialdehyde(MDA) in liver tissue homogenate were detected by using chromatoptometry. The concentration of serum c AMP and c GMP were detected by using radioimmunoassay(RIA),and ratio of c AMP to c GMP(c AMP/c GMP) was calculated. The activity of F0 F1-ATP enzyme was detected by using continuous cycle method,and activities of Na+-K+-ATP and Ca2 +-Mg2 +-ATP enzymes were detected by using enzymolysis. Results Compared with normal group,mouse body weight and time of FST decreased,activities of SOD,CAT and GSH-PX decreased,MDA content increased,c AMP/c GMP decreased,and activities of F0 F1-ATP,Na+-K+-ATP and Ca2 +-Mg2 +-ATP decreased in model group(P < 0. 01). Compared with model group,mouse body weight was increased in all Maka groups(P < 0. 01). The time of FST was significantly prolonged in high-dose Maka group and ginseng group(P < 0. 01),which was not significant in mid-dose and low-dose Maka groups. The activities of SOD,CAT and GSH-PX were significantly increased(P < 0. 01),MDA content was decreased(P < 0. 01),c AMP/c GMP was significantly increased(P < 0. 01),and activities of F0 F1-ATP,Na+-K+-ATP and Ca2 +-Mg2 +-ATP were significantly increased(P < 0. 01) in mid-dose and high-dose Maka groups,which was no significant in low-dose Maka group. Conclusion Maka,in a certain dose,can relieve physical fatigue,improve activities of anti-oxidative enzymes and eliminate cytotoxicity of free radicals,which verifies Maka's TCM properties of warming and fortifying spleen. The correlation between Maka's effective mechanism and enhancing mitochondrial energy metabolism enzymes is further confirmed.
引文
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[8]张广霞,赵静,刘碧原,等.不同健脾方对脾虚模型大鼠水盐代谢及水转运的作用比较[J].广州中医药大学学报,2016,33(1):51-55.Zhang GX,Zhao J,Liu BY,et al.Comparative study of effects of spleen-strengthening recipes on water-electrolyte metabolism and water transport in rats with spleen-deficiency syndrome[J].Journal of Guangzhou University of Traditional Chinese Medicine,2016,33(1):51-55.
[9]查圣华.玛咖活性成分及抗疲劳功能研究[D].北京:中国科学院大学,2016.Zha SH.Study on the active constituents of Maca and antifatigue function of Maca(Lepidium mevenii)[D].Beijing:University of Chinese Academy of Sciences,2016.
[10]杨泽民,陈蔚文.脾虚证与物质能量代谢紊乱相关性研究进展[J].广州中医药大学学报,2012,29(3):332-336.Yang ZM,Cheng WW.Research Progress on correlation between spleen deficiency pattern and material energy metabolism disorder[J].Journal of Guangzhou University of Traditional Chinese medicine,2012,29(3):332-336.
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[13]陈奇.c AMP、c GMP与中医阴阳学说[J].江西中医药,1980(2):12.Chen Q.c AMP,c GMP and yin-yang theory of TCM[J].Jiangxi Journal of Traditional Chinese Medicine,1980(2):12.
[14]张翠珍,王天芳.c AMP、c GMP拮抗性代谢调节与中医证候关系研究进展[J].北京中医药大学学报,1999,22(6):51-53.Zhang CZ,Wang TF.Research progress of relationship between antagonistic metabolic regulation of c AMP and c GMP and TCM patterns[J].Journal of Beijing University of Traditional Chinese Medicine,1999,22(6):51-53
[15]李峰,韩晨霞,吴凤芝,等.疲劳的现代研究[J].中国科学:生命科学,2016,46(8):903-912.Li F,Han CX,Wu FZ,et al.Modern research of fatigue[J].Scientia Sinica Vitae,2016,46(8):903-912.
[16]Oliveira TN,Possidonio AC,Soares CP,et al.The role of Na+/K+-ATPase during chick skeletal myogenesis[J].Plos One,2015,10(3):1-16.
[17]Boelens A D,Pradhan R K,Blomeyer C A,et al.Extramatrix Mg2+limits Ca2+uptake and modulates Ca2+uptake-independent respiration and redox state in cardiac isolated mitochondria[J].Journal of Bioenerg Biomembr,2013,45(3):203-218.
[18]张建军,王淳,费文婷,等.外来药物“中药化”的研究思路与策略[J].中华中医药杂志,2017,32(4):1425-1430.Zhang JJ,Wang C,Fei WT,et al.Strategies and approaches on introducing the foreign medicine to TCM[J].China Journal of Traditional Chinese Medicine and Pharmacy,2017,32(4):1425-1430.
[19]Gonzales GF,Gasco M,Lozada-requena I.Role of Maca(Lepidium meyenii)consumption on serum interleukin-6levels and health status in populations living in the Peruvian Central Andes over 4000 m of altitude[J].Plant Foods Hum Nutr,2013,68(4):347-351.
[20]Gonzales GF.Ethnobiology and Ethnopharmacology of Lepidium meyenii(Maca),a Plant from the Peruvian Highlands[J].Evidence-based Complementary and Alternative Medicine,2012(10):1-10.
[21]肖培根,刘勇,肖伟.西方草药概况及我国开发对策[J].中国药学杂志,2001,36(8):505-507.Xiao PG,Liu Y,Xiao W.General situation of Western herbal medicine and development strategy of China[J].Chin Pham J,2001,36(8):505-507.
[2]Melnikovova I,Fait T,Kolarova M,et al.Effect of Lepidium meyenii Walp.on semen parameters and serum hormone levels in healthy adult men:a double-blind,randomized,placebo-controlled pilot study[J].Evidence-Based Complementary and Alternative Medicine,2015,8(1):1-6.
[3]Meissner HO,Reich-Bilinska H,Mscisz A,et al.Therapeutic Effects of Pre-Gelatinized Maca(Lepidium Peruvianum Chacon)used as a Non-Hormonal Alternative to HRT in Perimenopausal Women-Clinical Pilot Study[J].International Journal of Biomedical Science,2006,2(2):143-159.
[4]肖培根,刘勇,肖伟.玛咖—全球瞩目的保健食品[J].国外医药·植物药分册,2001,16(6):236.Xiao PG,Liu Y,Xiao W.Maka-a globally recognized health food[J].World Notes on Plant Medicines,2001,16(6):236
[5]国家卫生健康委员会.关于批准玛咖粉作为新资源食品的公告(2011年第13号)[EB/OL].[2018-01-02].http://www.nhfpc.gov.cn/sps/s7891/201106/d18bef8a697e44da87f20eb-b41cb428e.shtml,2011-06-16.National Health Commission.Announcement on approval of Maca Powder as a new resource food(No.13 of 2011)[EB/OL].[2018-01-02]http://www.nhfpc.gov.cn/sps/s7891/201106/d18bef8a697e44da87f 20ebb41cb428e.shtml,2011-06-16.
[6]王淳,张建军,王佳,等.玛咖的临床文献研究及其中药性能探讨[J].中华中医药杂志,2017,32(12):5569-5573.Wang C,Zhang JJ,Wang J,et al.Clinical literature research and discussion of Chinese medicine properties of Maca[J].China Journal of Traditional Chinese Medicine and Pharmacy,2017,32(12):5569-5573
[7]刘玉,陈慧.脾虚病证结合动物模型研究进展[J].中国中医急症,2016,25(1):99-101.Liu Y,Chen H.Research progress of spleen deficiency pattern combined with animal model[j].J Emerg Tradit Chin Med,2016,25(1):99-101
[8]张广霞,赵静,刘碧原,等.不同健脾方对脾虚模型大鼠水盐代谢及水转运的作用比较[J].广州中医药大学学报,2016,33(1):51-55.Zhang GX,Zhao J,Liu BY,et al.Comparative study of effects of spleen-strengthening recipes on water-electrolyte metabolism and water transport in rats with spleen-deficiency syndrome[J].Journal of Guangzhou University of Traditional Chinese Medicine,2016,33(1):51-55.
[9]查圣华.玛咖活性成分及抗疲劳功能研究[D].北京:中国科学院大学,2016.Zha SH.Study on the active constituents of Maca and antifatigue function of Maca(Lepidium mevenii)[D].Beijing:University of Chinese Academy of Sciences,2016.
[10]杨泽民,陈蔚文.脾虚证与物质能量代谢紊乱相关性研究进展[J].广州中医药大学学报,2012,29(3):332-336.Yang ZM,Cheng WW.Research Progress on correlation between spleen deficiency pattern and material energy metabolism disorder[J].Journal of Guangzhou University of Traditional Chinese medicine,2012,29(3):332-336.
[11]吕爱平,李德新,刘春明,等.脾虚三证模型大鼠脂质过氧化损伤的比较[J].中国中西医结合消化杂志,2001,9(2):74-75.Lu AP,Li DX,Liu CM,et al.The Comparative research on damage of lipid peroxidation to spleen-qi deficiency,spleen-yang deficiency and spleen-yin deficiency in rats[J].Chinese Journal of Integrated Traditional and Western Medicine on Digestion,2001,9(2):74-75
[12]赵丽,张效莉,侯慧茹.脾虚小鼠过氧化与抗氧化改变的实验研究[J].辽宁中医药大学学报,2008,10(9):164-165.Zhao L,Zhang XL,Hou HR.Changes of peroxidation and antioxidation in mice with spleen deficiency[J].Journal of Liaoning University of Traditional Chinese Medicine,2008,10(9):164-165
[13]陈奇.c AMP、c GMP与中医阴阳学说[J].江西中医药,1980(2):12.Chen Q.c AMP,c GMP and yin-yang theory of TCM[J].Jiangxi Journal of Traditional Chinese Medicine,1980(2):12.
[14]张翠珍,王天芳.c AMP、c GMP拮抗性代谢调节与中医证候关系研究进展[J].北京中医药大学学报,1999,22(6):51-53.Zhang CZ,Wang TF.Research progress of relationship between antagonistic metabolic regulation of c AMP and c GMP and TCM patterns[J].Journal of Beijing University of Traditional Chinese Medicine,1999,22(6):51-53
[15]李峰,韩晨霞,吴凤芝,等.疲劳的现代研究[J].中国科学:生命科学,2016,46(8):903-912.Li F,Han CX,Wu FZ,et al.Modern research of fatigue[J].Scientia Sinica Vitae,2016,46(8):903-912.
[16]Oliveira TN,Possidonio AC,Soares CP,et al.The role of Na+/K+-ATPase during chick skeletal myogenesis[J].Plos One,2015,10(3):1-16.
[17]Boelens A D,Pradhan R K,Blomeyer C A,et al.Extramatrix Mg2+limits Ca2+uptake and modulates Ca2+uptake-independent respiration and redox state in cardiac isolated mitochondria[J].Journal of Bioenerg Biomembr,2013,45(3):203-218.
[18]张建军,王淳,费文婷,等.外来药物“中药化”的研究思路与策略[J].中华中医药杂志,2017,32(4):1425-1430.Zhang JJ,Wang C,Fei WT,et al.Strategies and approaches on introducing the foreign medicine to TCM[J].China Journal of Traditional Chinese Medicine and Pharmacy,2017,32(4):1425-1430.
[19]Gonzales GF,Gasco M,Lozada-requena I.Role of Maca(Lepidium meyenii)consumption on serum interleukin-6levels and health status in populations living in the Peruvian Central Andes over 4000 m of altitude[J].Plant Foods Hum Nutr,2013,68(4):347-351.
[20]Gonzales GF.Ethnobiology and Ethnopharmacology of Lepidium meyenii(Maca),a Plant from the Peruvian Highlands[J].Evidence-based Complementary and Alternative Medicine,2012(10):1-10.
[21]肖培根,刘勇,肖伟.西方草药概况及我国开发对策[J].中国药学杂志,2001,36(8):505-507.Xiao PG,Liu Y,Xiao W.General situation of Western herbal medicine and development strategy of China[J].Chin Pham J,2001,36(8):505-507.