The Anti-fatigue Effect And Its Mechanism Of Wild Ginseng Superfine Powder

Fatigue is a physiological phenomenon that often occurs in people’s daily lives.Usually,fatigue can be significantly recovered after rest.However,with the increasing working hours and intensity of people’s work,fatigue is often not effectively relieved in time,which may induce hypertension,angina pectoris,neurasthenia and other diseases and damage people’s health.At present,there are many kinds of anti-fatigue drugs,but their efficacy is not ideal and limited.It is important to develop a new drug to relieve human fatigue and protect human health.Ginseng（Panax ginseng C.A.Meyer）is a perennial herb of the genus Ginseng in the family of Wujia,with effects of blood pressure adjustment,heart function restoration,anti-nervous weakness and anti-fatigue,etc.In the National Standard of the People’s Republic of China"Identification and Grading Quality of Wild Mountain Ginseng"（GB/T 18765-2015）,ginseng sown artificially in wild mountain forests and grown naturally for more than 15 years is called"wild mountain ginseng".In this thesis,the ginseng grown naturally in the forest for 16 years（hereinafter referred to as wild ginseng）differs from garden ginseng in terms of total saponins and some monomeric saponins.Superfine powder has various advantages such as no decoction,easy to take,and originality.Wild Ginseng Superfine Powder（WGSP）is made by crushing raw sun-dried wild ginseng into a superfine powder with a diameter of less than 45μm.Objectives:The objective of this experimental study was to observe the effect of superfine powder of wild ginseng on antibody force fatigue in mice and its mechanism.Methods:1.Study on the antibody fatigue effect of wild ginseng superfine powder in mice1.1 Pole climbing testForty mice were randomly divided into control（Control）group and WGSP 100,200,400 mg/kg group according to body weight,10 mice in each group.Control group were given 0.5%CMC-Na 20 m L/kg by gavage（ig）,and the remaining groups were given the corresponding dose of drug 20 m L/kg by ig,1t/d,for 30 days.The body weight and food intake of the mice were measured and recorded before each day of administration;the pole climbing test train was started on the 28^th-29^th day,and the pole climbing test was conducted 1 h after the drug on day 30,and the time that the mice spent on the pole within 10 min was recorded as the pole climbing time.The heart,lung,liver and skeletal muscle of both hind legs were taken after the pole climbing,weighed and the organ indexes were calculated.1.2 Swimming exhaustion testThe grouping and drug administration methods were the same as 1.1.On the day 29^th,1 h after the drug,the mice were individually placed in a swimming tank（length′width′height=23′8′28 cm）with a water depth of 25 cm and a water temperature of 25-30℃,and swam without weight for 10 min.At 1 h after the drug on the 30^th day,the mice were loaded with a lead block of 6%of their own body weight on their tails,and the swimming exhaustion experiment was conducted.The exhaustion was judged by the mice’s head sinking into the water for 10 s without surfacing,and the swimming exhaustion time was recorded.1.3 Treadmill fatigue testThe grouping and drug administration methods were the same as 1.1.On the day 27^th-29^th,1 h after drug administration,the mice were trained to run.On the 30^th day,1 h after drug administration,the mice were tested by running,the running speed was started at 14 m/min,and the running speed was increased by 2 m/min,and it was not increased when the speed reached 34 m/min.The mice were judged to be fatigued when they stayed in the"fatigue zone"for more than 5 s at a body length from the electric shock net.The time from the start of running to exhaustion and the running distance were recorded.1.4 Determination of blood lactate content,muscle glycogen and liver glycogen contentThe grouping and drug administration methods were the same as 1.1.On the 30^th day,1 h after the drug,the mice were loaded with 5%of their own body weight on the tail and swam for 10 min,then dried after swimming and put back into the original cage for 20 min.Venous blood was collected before swimming,10 min after swimming and 20 min after resting,and the lactic acid（LA）content of mice was measured and the area under the curve of lactic acid change was calculated.Liver and skeletal muscle of the left hind limb were collected,and biochemical kits were used to detect muscle glycogen and liver glycogen.1.5 Determination of serum urea nitrogen contentThe grouping and drug administration methods were the same as 1.1.On the 30^th day,1 h after the drug,the mice were individually placed in a swimming bucket（r=12 cm）with water depth of 25 cm and water temperature of 25-30℃,and swam without weight for 90 min,dried after swimming,and put back into the original feeding cage for 60 min resting.After resting,blood was taken from the eye,and the serum was measured by biochemical kit for serum urea nitrogen（BUN）content.2.Study on the mechanism of antibody fatigue effect of wild ginseng superfine powder2.1 Detection of oxidative stress injury-related indexesThe grouping and drug administration methods were the same as 1.1.On the 30^th day,1 h after the drug,mice were put into a swimming tank（length′width′height=23′8′28 cm）with 25 cm water depth and 25-30℃water temperature,and swam for 10 min with 5%of their own body weight on the tail.After swimming for 10 min,blood was taken from the eye and skeletal muscle was taken from the left hind limb of the mice.The biochemical kits were used to detect lactate dehydrogenase（LDH）and creatine kinase（CK）activities in serum,total superoxide dismutase（T-SOD）activity and malondialdehyde（MDA）content in muscle tissue.2.2 Detection of ATPase activity and ATP content in liver and skeletal muscleLiver tissues and skeletal muscles of the right hind limbs of the mice in 2.1experiments were taken.Biochemical kits were used to detect Ca²⁺-Mg²⁺-ATPase and Na⁺-K⁺-ATPase activities in muscle and liver tissues and ATP content in muscle tissues.2.3 Expression levels of m RNA of skeletal muscle mitochondrial oxidative respiratory chain-related proteinsThe expression levels of mitochondrial oxidative respiratory chain complex I-V were measured by q RT-PCR in the skeletal muscle of the right hind limb of 1.2experimental mice.2.4 Skeletal muscle mitochondrial mt DNA copy number detectionThe mitochondrial mt DNA copy number was measured by q RT-PCR in the skeletal muscle of the right hind limb of 1.2 mice.2.5 Detection of AMPK/PGC-1αsignaling pathway-related proteins in skeletal muscleThe expression levels of AMPK/PGC-1αsignaling pathway-related proteins in skeletal muscle were detected by Western Blot from the right hind limb of mice in 1.2.Results:1.Study on the effect of wild ginseng superfine powder against physical fatigue in mice1.1 Compared with Control group,the daily food intake,average weekly body weight and heart,lung,liver and skeletal muscle organ coefficients of mice in WGSP100,200 and 400 mg/kg groups were not significantly changed（P>0.05）,and WGSP200 and 400 mg/kg could significantly prolong the pole climbing time of mice（P<0.05）.1.2 Compared with the Control group,WGSP 200 and 400 mg/kg could significantly prolong the time of swimming exhaustion in mice（P<0.05 or P<0.01）.1.3 Compared with the Control group,WGSP 200 and 400 mg/kg could significantly increase the running time and running distance of mice（P<0.05 or P<0.01）.1.4 Compared with the Control group,the area under the lactate change curve of mice in the WGSP 100,200 and 400 mg/kg groups was significantly reduced,muscle glycogen content was significantly reduced and liver glycogen content was significantly increased（P<0.05 or P<0.01）.1.5 Compared with the Control group,the BUN content of mice in WGSP 100,200 and 400 mg/kg groups was significantly reduced（P<0.01）.2.Study on the mechanism of action of wild ginseng superfine powder against force fatigue2.1 Compared with the Control group,the skeletal muscle T-SOD activity of mice in the WGSP 200 and 400 mg/kg groups increased significantly and the MDA content decreased significantly（P<0.05 or P<0.01）;compared with the Control group,the serum LDH activity of mice in the WGSP 100,200 and 400 mg/kg groups decreased significantly（P<0.05 or P<0.01）,and serum CK activity was significantly reduced in mice of WGSP 200 and 400 mg/kg groups（P<0.05 or P<0.01）.2.2 Compared with the Control group,the activities of Ca²⁺-Mg²⁺-ATPase and Na⁺-K⁺-ATPase in liver tissues of mice in the WGSP 100,200 and 400 mg/kg groups were not significantly changed（P>0.05）,and the activities of Ca²⁺-Mg²⁺-ATPase and Na⁺-K⁺-ATPase in muscle tissues were significantly increased（P<0.05 or P<0.01）,and the skeletal muscle ATP content was significantly increased in the WGSP 200 and 400mg/kg groups of mice（P<0.01）.2.3 Compared with Control group,MT-CO1,MT-CO2 and MT-CO3 m RNA levels were significantly increased in skeletal muscle of mice in WGSP 100,200 and 400mg/kg groups（P<0.01）,and COX4il,ATP5a1 m RNA levels were significantly increased in skeletal muscle of mice in WGSP 200 and 400 mg/kg groups（P<0.01）.2.4 The expression of mitochondrial DNA marker gene D-loop was significantly increased in skeletal muscle of mice in the WGSP 200 and 400 mg/kg groups compared with the Control group（P<0.01）.2.5 The expression of p-AMPK,PGC-1α,NRF1 and TFAM proteins were significantly increased in skeletal muscle of mice in the WGSP 100,200 and 400 mg/kg groups compared with the Control group（P<0.05 or P<0.01）.Conclusion:The results of this study showed that WGSP significantly increased swimming exhaustion time,pole climbing time,running time and running distance,decreased LA and BUN content,increased mitochondrial number and enhanced skeletal muscle mitochondrial function,and reduced exercise injury in mice,and its effects may be related to activation of AMPK/PGC-1αsignaling pathway.