火把花根多糖对运动疲劳小鼠AMPK/mTOR信号通路的影响

doi:10.11958/20220320

天津医药 ›› 2022, Vol. 50 ›› Issue (11): 1165-1170.doi: 10.11958/20220320

火把花根多糖对运动疲劳小鼠AMPK/mTOR信号通路的影响

杜伟¹(), 吴思澜², 杨雪², 陈慧², 叶兰¹^,^△()

1 贵州医科大学基础医学院药理学教研室（邮编550025）
2 重庆市中药研究院

收稿日期:2022-02-28 修回日期:2022-04-10 出版日期:2022-11-15 发布日期:2022-11-11
通讯作者: 叶兰 E-mail:361521169@qq.com;frogyl266@163.com
作者简介:杜伟（1988），男，硕士在读，主要从事新药药理毒理学方面研究。E-mail：361521169@qq.com
基金资助:
重庆市科技局资助项目(cstc2021jcyj-msxmX0808)

Effects of tripterygium hypoflaucum (Levl.) hutch polysaccharide on the AMPK/mTOR signal pathway in mice with exercise-induced fatigue

DU Wei¹(), WU Silan², YANG Xue², CHEN Hui², YE Lan¹^,^△()

1 Department of Pharmacology, School of Basic Science, Guizhou Medical University, Guiyang 550025, China
2 Chongqing Academy of Chinese Materia Medica

Received:2022-02-28 Revised:2022-04-10 Published:2022-11-15 Online:2022-11-11
Contact: YE Lan E-mail:361521169@qq.com;frogyl266@163.com

杜伟, 吴思澜, 杨雪, 陈慧, 叶兰. 火把花根多糖对运动疲劳小鼠AMPK/mTOR信号通路的影响[J]. 天津医药, 2022, 50(11): 1165-1170.

DU Wei, WU Silan, YANG Xue, CHEN Hui, YE Lan. Effects of tripterygium hypoflaucum (Levl.) hutch polysaccharide on the AMPK/mTOR signal pathway in mice with exercise-induced fatigue[J]. Tianjin Medical Journal, 2022, 50(11): 1165-1170.

摘要/Abstract

摘要：

目的研究火把花根多糖的抗运动疲劳作用及可能的作用机制。方法 24只ICR雄性小鼠按随机数字表法分为静止对照组、疲劳对照组、疲劳+单糖组和疲劳+火把花根多糖组，每组6只。疲劳+火把花根多糖组给予20 g/L火把花根多糖受试液，疲劳+单糖组给予20 g/L单糖混合物受试液，静止对照组和疲劳对照组给予纯化水，各组均按20 mL/kg每日灌胃给药1次，连续14 d。观察给药期间小鼠体质量、摄食量、饮水量等一般情况；在给药第6、13天进行疲劳转棒实验，记录小鼠疲劳转棒时间；给药第14天进行力竭游泳实验，记录小鼠力竭游泳时间；小鼠力竭游泳后即相应取材进行血清的三酰甘油（TG）、血糖（GLU）检测，行左侧腓肠肌HE染色和糖原（PAS）染色观察病理变化，右侧腓肠肌采用实时荧光定量PCR（qPCR）方法检测腺苷酸活化蛋白激酶（AMPK）、哺乳动物雷帕霉素靶蛋白（mTOR）、葡萄糖转运蛋白4（GLUT-4）mRNA表达水平。结果给药第13天，相比静止对照组，疲劳+火把花根多糖组的体质量减轻（P<0.05）。其余各时间点各组的体质量、摄食量、饮水量的差异均无统计学意义（P>0.05）。相比疲劳对照组，疲劳+火把花根多糖组在给药第6天时疲劳转棒时间增加不显著（P>0.05），而在给药第13天时，其疲劳转棒时间显著增加（P<0.05），且在给药第14天时其力竭游泳时间亦增加（P<0.05）。HE染色未见明显异常，PAS染色可见疲劳对照组与疲劳+单糖组腓肠肌着色略深，而疲劳+火把花根多糖组着色最深。与疲劳对照组比较，疲劳+火把花根多糖组的TG差异无统计学意义，GLU降低，而AMPK、mTOR、GLUT-4 mRNA表达水平显著上调（P<0.05）。结论火把花根多糖具有抗运动疲劳的作用，其机制可能与调节AMPK/mTOR信号通路的糖代谢有关。

关键词: 运动, 疲劳综合征，慢性, 游泳, 多糖类, 火把花, AMPK/mTOR信号通路

Abstract:

Objective To study the effect and possible mechanism of polysaccharide from tripterygium hypoflaucum (Levl.) hutch on exercise-induced fatigue in mice. Methods Twenty-four ICR mice were randomly divided into the static control group, the fatigue control group, the fatigue + monosaccharide group and the fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group, with 6 mice in each group. The fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group was given 20 g/L aqueous solution of tripterygium hypoflaucum (Levl.) hutch polysaccharide, the fatigue + monosaccharide group was given 20 g/L aqueous solution of monosaccharide mixture, and the static control group and the fatigue control group were given purified water. Each group was given intervention 20 mL/kg by gavage once a day for 14 days. The body mass, food intake, water consumption and other general conditions of mice were observed during administration. On the 6th day and the 13th day of administration, mice were evaluated by rotary rod fatigue tester, and on the 14th day of administration, the exhaustive swimming experiment was carried out. After exhaustive swimming, the serum levels of triglyceride (TG) and blood glucose (GLU) were detected. The pathological changes of left gastrocnemius muscle were observed by HE staining and Periodic Acid-Schiff staining (PAS). The expression levels of AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR) and glucose transporter 4 (GLUT-4) mRNA were observed by real-time fluorescence quantitative PCR (qPCR). Results After treatment for 13 days, compared with the static control group, the body mass was decreased in the fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group (P<0.05). There were no significant differences in body mass, food intake and water consumption at other time points between the four groups (P>0.05). Compared with the fatigue control group, there was no significant increase in the rotating rod lasting time after treatment for 6 days in the fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group (P>0.05), but it increased significantly on the 13th day of administration (P<0.05). The exhaustive swimming time significant increased after treatment for 14 days in the fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group (P<0.05). HE staining showed no obvious abnormality, PAS staining showed that the gastrocnemius muscle was slightly darker in the fatigue control group and the fatigue + monosaccharide group, while the colour was the deepest in the fatigue+tripterygium hypoflaucum (Levl.) hutch polysaccharide group. Compared with the fatigue control group, there was no significant difference in TG, the GLU decreased in the fatigue + tripterygium hypoflaucum (Levl.) hutch polysaccharide group (P<0.05), while the expression levels of AMPK, mTOR and GLUT-4 mRNA were significantly increased (P<0.05). Conclusion The tripterygium hypoflaucum (Levl.) hutch polysaccharide has the effect of anti-exercise-induced fatigue in mice, and its mechanism may be related to the glucose metabolism regulated by AMPK/mTOR signal pathway.

Key words: exercise, fatigue syndrome, chronic, swimming, polysaccharides, tripterygium hypoflaucum (Levl.) hutch, AMPK/mTOR signaling pathway

中图分类号:

R285.5

图/表 8

参考文献 23

[1]	王爱霞, 崔胜文, 丁玉婵. 霍山石斛运动饮料研制及其抗疲劳功能研究[J]. 食品安全质量检测学报, 2022, 13(1):248-253.
	WANG A X, CUI S W, DING Y C. Study on the development of dendrobium huoshanense sprorts drink and its anti-fatigue function[J]. Journal of Food Safety & Quality, 2022, 13(1):248-253.
[2]	VAN'T LEVEN M, ZIELHUIS G A, VAN DER MEER J W, et al. Fatigue and chronic fatigue syndrome-like complaints in the general population[J]. European Journal of Public Health, 2010, 20(3):251-257. doi:10.1093/eurpub/ckp113.
[3]	伍侨, 高静, 柏丁兮, 等. 中国人群慢性疲劳综合征患病率的Meta分析[J]. 右江医学, 2020, 48(10):727-735.
	WU Q, GAO J, BAI D X, et al. Prevalence of chronic fatigue syndrome in China:A meta-analysis[J]. Youjiang Medical Journal, 2020, 48(10):727-735. doi:10.3969/j.issn.1003-1383.2020.10.002.
[4]	LUO C, XU X, WEI X, et al. Natural medicines for the treatment of fatigue:Bioactive components,pharmacology,and mechanisms[J]. Pharmacological Research, 2019, 148:104409. doi:10.1016/j.phrs.2019.104409.
[5]	ZHANG L J, FU M, CHEN J L, et al. Supplementation with embryo chicken egg extract improves exercise performance and exerts anti-fatigue effects via AMPK/mTOR signalling pathway in mice[J]. J Sci Food Agric, 2021, 101(4):1411-1418. doi:10.1002/jsfa.10754.
[6]	KJØBSTED R, HINGST J R, FENTZ J, et al. AMPK in skeletal muscle function and metabolism[J]. The FASEB Journal, 2018, 32(4):1741-1777. doi:10.1096/fj. 201700442R.
[7]	ZHOU S S, JIANG J G. Anti-fatigue effects of active ingredients from traditional Chinese Medicine:A review[J]. Curr Med Chem, 2019, 26(10):1833-1848. doi:10.2174/0929867324666170414164607.
[8]	ZHANG C J, GUO J Y, CHENG H, et al. Spatial structure and anti-fatigue of polysaccharide from Inonotus obliquus[J]. Int J Biol Macromol, 2020, 151:855-860. doi:10.1016/j.ijbiomac.2020.02.147.
[9]	FENG Q, SI Y, ZHU L L, et al. Anti-inflammatory effects of a SERP 30 polysaccharide from the residue of Sarcandra glabra against lipopolysaccharide-induced acute respiratory distress syndrome in mice[J]. J Ethnopharmacol, 2022, 293:115262. doi:10.1016/j.jep.2022.115262.
[10]	郑传痴, 杨艳, 韦余, 等. 金丝桃苷对小鼠的抗疲劳作用及机制研究[J]. 食品工业科技, 2021, 42(23):350-355.
	ZHENG C C, YAN Y, WEI Y, et al. Study on the effects and mechanism of Hyperoside on anti-fatigue in mice[J]. Science and Technology of Food Industry, 2021, 42(23):350-355. doi:10.13386/j.issn1002-0306.2021010227.
[11]	YANG Z, SUNIL C, JAYACHANDRAN M, et al. Anti-fatigue effect of aqueous extract of hechong(Tylorrhynchus heterochaetus)via AMPK linked pathway[J]. Food Chem Toxicol, 2020, 135:111043. doi:10.1016/j.fct.2019.111043.
[12]	井宏颖, 吕克宁, 宋晓晨, 等. 地黄饮子对运动性疲劳小鼠运动能力的影响[J]. 湖南中医药大学学报, 2021, 41(1):34-38.
	JING H Y, LV K N, SONG X C, et al. Effect of radix rehmanniae decoction on exercise ability in exercise-induced fatigue mice[J]. Journal of Traditional Chinese Medicine University of Hunan, 2021, 41(1):34-38. doi:10.3969/j.issn.1674-070X.2021.01.007.
[13]	YU J, LAYBUTT D R, KIM L J, et al. Exercise-induced benefits on glucose handling in a model of diet-induced obesity are reduced by concurrent nicotinamide mononucleotide[J]. AJP Endocrinology and Metabolism, 2021, 321(1):176-189. doi:10.1152/ajpendo.00446.2020.
[14]	TIAN L, HU T, ZHANG S S, et al. A comparative study on relieving exercise-induced fatigue by inhalation of different citrus essential oils[J]. Molecules, 2022, 27:3239. doi:10.3390/molecules27103239.
[15]	YIN C, FU X, CHOU J, et al. A proprietary herbal drug Young Yum Pill ameliorates chronic fatigue syndrome in mice[J]. Phytomedicine, 2021, 88:153602. doi:10.1016/j.phymed.2021.153602.
[16]	HU B, YE C, LEUNG E L, et al. Bletilla striata oligosaccharides improve metabolic syndrome through modulation of gut microbiota and intestinal metabolites in high fat diet-fed mice[J]. Pharmacol Res, 2020, 159:104942. doi:10.1016/j.phrs.2020.104942.
[17]	姜宁, 张亦文, 姚彩虹, 等. 大小鼠抑郁行为实验方法概述[J]. 中国实验动物学报, 2021, 29(6):830-838.
	JIANG N, ZHANG Y W, YAO C H, et al. Overview of animal behavioral tests of depression[J]. Acta Laboratorium Animalis Scientia Sinica, 2021, 29(6):830-838. doi:10.3969/j.issn.1005-4847.2021.06.016.
[18]	董碧莲, 蔡延渠, 吕莉, 等. 中药多糖增强免疫,抗疲劳作用的研究进展[J]. 中成药, 2019, 41(5):1119-1124.
	DONG B L, CAI Y Q, LV L, et al. Research progress of traditional Chinese medicine polysaccharide in enhancing immunity and anti fatigue[J]. Chinese Traditional Patent Medicine, 2019, 41(5):1119-1124. doi:10.3969/j.issn.1001-1528.2019.05.032.
[19]	OSMAN W, MOHAMED S. Standardized morinda citrifolia L. and Morinda elliptica L. leaf extracts alleviated fatigue by improving glycogen storage and lipid/carbohydrate metabolism[J]. Phytother Res, 2018, 32(10):2078-2085. doi:10.1002/ptr.6151.
[20]	MORALES-ALAMO D, CALBET J A L. AMPK signaling in skeletal muscle during exercise:Role of reactive oxygen and nitrogen species[J]. Free Radic Biol Med, 2016, 98(9):68-77. doi:10.1016/j.freeradbiomed.2016.01.012.
[21]	GONZÁLEZ A, HALL M N, LIN S C, et al. AMPK and TOR:The Yin and Yang of cellular nutrient sensing and growth control[J]. Cell Metab, 2020, 31(3):472-492. doi:10.1016/j.cmet.2020.01.015.
[22]	张新颖, 毛景东, 杨晓燕, 等. AMPK/mTOR信号通路的研究进展[J]. 微生物学杂志, 2019, 39(3):109-116.
	ZHANG X Y, MAO J D, YANG X Y. Advances in AMPK/mTOR signaling path[J]. Journal of Microbiology, 2019, 39(3):109-116. doi:10.3969/j.issn.1005-7021.2019.03.015.
[23]	LI J, KNUDSEN J R, HENRIQUEZ-OLGUIN C, et al. AXIN1 knockout does not alter AMPK/mTORC1 regulation and glucose metabolism in mouse skeletal muscle[J]. The Journal of Physiology, 2021, 599(12):3081-3100. doi:10.1113/JP281187.

组别	第1天	第4天	第7天	第10天	第13天
静止对照组	20.1±0.6	24.0±1.2	27.3±1.9	30.0±2.3	31.4±2.0
疲劳对照组	20.1±0.8	24.1±1.4	27.4±2.0	30.1±2.2	31.4±2.4
疲劳+单糖组	20.1±0.7	24.1±1.0	27.3±1.6	29.1±1.4	29.8±1.4
疲劳+火把花根多糖组	20.1±0.7	23.5±1.1	25.8±1.5	27.5±1.9	28.1±1.7^a
F	0.003	0.357	1.141	2.119	4.192^＊

组别	第1天	第4天	第7天	第10天	第13天
静止对照组	20.1±0.6	24.0±1.2	27.3±1.9	30.0±2.3	31.4±2.0
疲劳对照组	20.1±0.8	24.1±1.4	27.4±2.0	30.1±2.2	31.4±2.4
疲劳+单糖组	20.1±0.7	24.1±1.0	27.3±1.6	29.1±1.4	29.8±1.4
疲劳+火把花根多糖组	20.1±0.7	23.5±1.1	25.8±1.5	27.5±1.9	28.1±1.7^a
F	0.003	0.357	1.141	2.119	4.192^＊

组别	第1天	第4天	第7天	第10天	第13天
静止对照组	9.50±1.25	12.00±0.90	12.42±0.80	12.67±0.78	12.00±0.82
疲劳对照组	9.37±0.70	11.73±0.70	12.77±0.91	12.73±0.74	12.17±0.35
疲劳+单糖组	9.40±1.04	12.33±0.91	12.60±0.79	12.68±1.12	11.80±0.66
疲劳+火把花根多糖组	9.50±0.95	12.50±0.89	12.37±1.12	12.50±1.48	12.03±0.60
F	0.014	0.483	0.115	0.026	0.174

组别	第1天	第4天	第7天	第10天	第13天
静止对照组	9.50±1.25	12.00±0.90	12.42±0.80	12.67±0.78	12.00±0.82
疲劳对照组	9.37±0.70	11.73±0.70	12.77±0.91	12.73±0.74	12.17±0.35
疲劳+单糖组	9.40±1.04	12.33±0.91	12.60±0.79	12.68±1.12	11.80±0.66
疲劳+火把花根多糖组	9.50±0.95	12.50±0.89	12.37±1.12	12.50±1.48	12.03±0.60
F	0.014	0.483	0.115	0.026	0.174

组别	第1天	第4天	第7天	第10天	第13天
静止对照组	11.6±0.6	14.8±0.8	16.1±0.6	14.9±0.9	13.3±0.7
疲劳对照组	11.7±0.9	15.2±0.4	15.9±0.4	15.0±0.2	13.1±0.4
疲劳+单糖组	12.0±0.7	15.2±0.4	16.2±0.7	15.0±0.7	13.2±0.9
疲劳+火把花根多糖组	12.1±0.2	14.8±0.9	15.8±0.5	14.9±0.9	13.4±0.3
F	0.354	0.313	0.297	0.015	0.173

火把花根多糖对运动疲劳小鼠AMPK/mTOR信号通路的影响

Effects of tripterygium hypoflaucum (Levl.) hutch polysaccharide on the AMPK/mTOR signal pathway in mice with exercise-induced fatigue

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参考文献 23

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基因名称	引物序列（5'→3'）	产物大小（bp）
AMPK	上游：GTTGCGGATGCTATGGGT	101
AMPK	下游：GTCCGGCCTCTGGTTTC	101
mTOR	上游：CTGGGGCTGCTTTCTGT	119
mTOR	下游：ACGGTTTTCTGCCTCTTGT	119
GLUT-4	上游：GTTGCGGATGCTATGGGT	104
GLUT-4	下游：GTCCGGCCTCTGGTTTC	104
GAPDH	上游：GGATGCTGCCCTTACCC	101
GAPDH	下游：GTTCACACCGACCTTCACC	101

组别	n	疲劳转棒时间		力竭游泳时间
组别	n	第6天	第13天	力竭游泳时间
疲劳对照组	6	2.74±1.10	2.63±1.45	10.2±3.4
疲劳+单糖组	6	2.66±1.05	2.84±1.76	14.0±3.3
疲劳+火把花根多糖组	6	4.12±0.96	10.58±2.36^a	24.2±4.8^a
F		3.737	34.268^＊＊	20.512^＊＊

组别	n	TG	GLU
静止对照组	6	0.67±0.11	6.33±1.52
疲劳对照组	6	1.08±0.36^a	3.35±1.11^a
疲劳+单糖组	6	1.05±0.13^a	2.75±1.08^a
疲劳+火把花根多糖组	6	1.06±0.15^a	1.40±0.40^ab
F		5.111^＊＊	21.405^＊＊

组别	AMPK mRNA	mTOR mRNA	GLUT-4 mRNA
静止对照组	1.13±0.59	1.18±0.62	1.09±0.50
疲劳对照组	1.34±0.57	1.14±0.90	1.13±0.83
疲劳+单糖组	1.51±0.51	1.57±0.87	1.08±0.28
疲劳+火把花根多糖组	2.20±0.82^ab	2.62±0.95^ab	2.90±0.58^ab
F	3.181^＊	4.007^＊	14.381^＊＊

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