探讨保元汤加减方通过AMPK/SIRT1/PGC－1α通路对小鼠的抗疲劳作用
Study on the anti-fatigue effect of modified
Baoyuan Decoction through the AMPK/SIRT1/PGC-1α pathway- 北京中医药大学学报 2023年46卷第12期 页码：1716-1727
- 作者机构：
1.浙江中医药大学药学院 杭州 310053
2.浙江工业大学
- 作者简介：
魏柯健，男，在读硕士生
#吕圭源，男，教授，博士生导师，主要研究方向：中药药理与新产品开发，E-mail：zjtcmlgy@163.com
- 基金信息：浙江省重点研发计划项目(2020C04020)
DOI：10.3969/j.issn.1006-2157.2023.12.012
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魏柯健, 俞静静, 苏洁, 等. 探讨保元汤加减方通过AMPK/SIRT1/PGC－1α通路对小鼠的抗疲劳作用[J]. 北京中医药大学学报, 2023,46(12):1716-1727.
WEI Kejian, YU Jingjing, SU Jie, et al. Study on the anti-fatigue effect of modified
Baoyuan Decoction through the AMPK/SIRT1/PGC-1α pathway[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(12):1716-1727.魏柯健, 俞静静, 苏洁, 等. 探讨保元汤加减方通过AMPK/SIRT1/PGC－1α通路对小鼠的抗疲劳作用[J]. 北京中医药大学学报, 2023,46(12):1716-1727. DOI： 10.3969/j.issn.1006-2157.2023.12.012.
WEI Kejian, YU Jingjing, SU Jie, et al. Study on the anti-fatigue effect of modified
Baoyuan Decoction through the AMPK/SIRT1/PGC-1α pathway[J]. Journal of Beijing University of Traditional Chinese Medicine, 2023,46(12):1716-1727. DOI： 10.3969/j.issn.1006-2157.2023.12.012.
摘要
目的,2,探讨保元汤加减方(人参、黄芪、甘草、肉桂、红景天)缓解疲劳的作用及机制。,方法,2,用HPLC检测保元汤加减方主要成分及含量。将160只SPF级ICR雄性小鼠分为4部分(各40只)，每部分均完全随机分为空白对照组(等体积纯化水)及保元汤加减方低、中、高剂量(1.9、2.1、2.3 g/kg)组，各组每日灌胃1次，共30 d。末次灌胃后取材并处死。(1)40只小鼠分别于第21天检测行为学指标(抓力、背温、自主活动次数、转棒疲劳时间)，第30天检测负重游泳时间。(2)40只小鼠用于检测肝/肌糖原水平。(3)40只用于检测血乳酸(BLA)含量及计算血乳酸曲线下面积。(4)40只小鼠用于检测机体代谢产物[血清尿素氮(BUN)、乳酸脱氢酶(LDH)、BLA]水平，氧化应激相关酶[骨骼肌及心脏中丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)],活性，血液运氧能力[全血红细胞(RBC)、血红蛋白(HGB)、平均血红蛋白浓度(MCHC)]，三磷酸腺苷(ATP)相关酶[骨骼肌及心脏中Na,＋,-K,＋,-ATP酶、Ca,2＋,-Mg,2＋,-ATP酶、琥珀酸脱氢酶(SDH)及血清中肌酸激酶(CK)]活性，腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子2相关酶1(SIRT1)/过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)通路相关蛋白表达。,结果,2,保元汤加减方中人参皂苷Rg,1,、人参皂苷Re、人参皂苷Rb,1,、红景天苷、毛蕊异黄酮含量分别为8.15、3.82、1.07、0.41、0.067 g/L。各剂量保元汤加减方可增加心脏与骨骼肌中Na,＋,-K,＋,-ATP酶、Ca,2＋,-Mg,2＋,-ATP酶、SDH、CAT、SOD活性(均,P,<,0.05)；降低小鼠心脏与骨骼肌中MDA水平(均,P,<,0.05)；降低小鼠血清中CK、LDH水平(均,P,<,0.05)；增加肝糖原、肌糖原储备量(均,P,<,0.05)；降低小鼠血清BUN水平与BLA曲线下面积(均,P,<,0.05)；增加全血RBC、HGB与MCHC(均,P,<,0.05)；增加小鼠骨骼肌和心脏p-AMPK、SIRT1、PGC-1α、TFAM蛋白表达(均,P,<,0.05)。,结论,2,保元汤加减方可能通过激活骨骼肌、心脏中AMPK/SIRT1/PGC-1α信号通路，增强线粒体能力，减少机体代谢产物的积累，抑制机体氧化应激与增加血液运氧能力，达到抗疲劳的作用。
Abstract
Objective,2,We modified ,Baoyuan, Decoction(ginseng, milkvetch root, liquorice root, cassia bark, and rose-boot) based on the original formula and explored its alleviating effect on physical fatigue.,Methods,2,Modified ,Baoyuan, Decoction (MBYD) were analyzed by using HPLC. Altogether 160 ICR mice with SPF grade were divided into 4 parts(,n,=40 mice per part), in which were divided into the blank control group (purifed water in the same volume), the modified ,Baoyuan, Decoction low-, mid-, and high-dose groups (1.9, 2.1, and 2.3 g/kg). All the groups were administered intragatrically once a day for 30 days. The mice were sacrificed and sampled after the last administration. (1) 40 mice were used to get the behavioral indicators. After 21 days of administration, grip strength, back temperature, times of locomotor activity, and fatigue time of rotating rod was measured; while time of weight-loading swimming was measured at the 30th day. (2) 40 mice were used to get liver/muscle glycogen levels. (3) 40 mice were used to get the content of blood lactic acid (BLA) and its area under the curve (AUC). (4) 40 mice were used to get (ⅰ) the levels of the metabolic products (blood urea nitrogen, BUN; lactate dehydrogenase, LDH; BLA); (ⅱ) activity of the oxidative stress-related enzymes (malondialdehyde, MDA; superoxide dismutase, SOD; catalase, CAT) in the skeletal muscle and the heart; (ⅲ)blood oxygen transport capacity (red blood cell, RBC; hemoglobin, HGB; mean corpuscular hemoglobin concentration, MCHC); (ⅳ) activity of ATP-related enzymes(Na,+, -K,+, -ATPase; Ca,2+, -Mg,2+, -ATPase; succinate dehydrogenase, SDH; creatine kinase, CK); (ⅴ) protein expression of phosphorylation alpha-AMP-activated protein kinase (p-AMPK), slient mating type information regulation 2 homolog 1(SIRT1), peroxisome proliferator activated receptor γ coactivator-1α (PGC-1α) and mitochondrial transcription factor A(TFAM).,Results,2,The contents of ginsenoside Rg,1, ginsenoside Re, ginsenoside Rb,1, salideoside and calycosin in MBYD were 8.15, 3.82, 1.07, 0.41 and 0.067 g/L. MBYD can significantly improve the activity of Na,+, -K,+, -ATPase, Ca,2+, Mg,2+, -ATPase, SDH, CAT, and SOD in the heart and skeletal muscles (all ,P,<,0.05); reduce the MDA levels in the heart and skeletal muscles(both ,P,<,0.05); reduce the levels of CK and LDH in serum (both ,P,<,0.05); increase liver glycogen and muscle glycogen reserves(both ,P,<,0.05); reduce the BUN level and the AUC of BLA (all ,P,<,0.05); increase the numbers of RBC, HGB, and MCHC in whole blood (all ,P,<,0.05); and increase the protein expression levels of p-AMPK, SIRT1, PGC-1α, and TFAM in skeletal muscle and heart (all ,P,<,0.05).,Conclusion,2,Modified ,Baoyuan, Decoction may activate the AMPK/SIRT1/PGC-1α signaling pathway in skeletal muscle and heart, enhance mitochondrial capacity, reduce the accumulation of metabolic products in the body, inhibit oxidative stress, and increase blood oxygen transport capacity, thereby alleviating fatigue.
关键词
保元汤补气抗疲劳线粒体腺苷酸活化蛋白激酶/沉默信息调节因子2相关酶1/过氧化物酶体增殖物激活受体γ辅激活因子1α通路小鼠
Keywords
Baoyuan Decoctiontonify qianti-fatiguemitochondrionalpha-AMP-activated protein kinase/slient mating type information regulation 2 homolog 1/peroxisome proliferator activated receptor γ coactivator-1α signal pathwaymice
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