CN106265703A

CN106265703A - Ginsenoside Rg1's application in preparation improves body movement ability health product

Info

Publication number: CN106265703A
Application number: CN201510233880.1A
Authority: CN
Inventors: 霍洪亮; 贾胜男; 李凤玉; 彭雪微; 王平; 于清淼; 孙丽丽
Original assignee: Northeast Normal University
Current assignee: Northeastern University China; Northeast Normal University
Priority date: 2015-05-11
Filing date: 2015-05-11
Publication date: 2017-01-04

Abstract

The invention belongs to technical field of bioengineering, be specifically related to ginsenoside Rg1's application in preparation improves body movement ability health product.

Description

Ginsenoside Rg1's application in preparation improves body movement ability health product

Technical field

Background technology

Raising combatant, rescue and relief work personnel, high intensity sports personnel and the motor capacity participating in various athletic competition animal are mankind's dreams for a long time.In the past, people mainly improved motor capacity by extra-nutrition material and reinforcement physical training.Showing according to the investigation of United States Olympic Committee in 2004, the athlete that there are about more than 90% supplements all kinds of nutrient substance by diet.This phenomenon the most generally exists in other high intensity muscle power staff.Nutrient substance mends sugar, salt from previously, to supplementing aminoacid, creatine and taurine in recent years, even with steroid hormone and analeptic.It is true that some nutrient substance of excessive use may bring a lot of potential risks and harm.Supplementing hormone and analeptic are played foul and relevant law especially.Along with progress and the development of science and technology, people have gradually recognized the contractile mechanism of muscle and the reason that muscle generation is tired, have recognized the mutual relation of muscle fatigue and physical fatigue.Skeletal muscle efficiency operation and the effective ways of resisting fatigue are promoted it has also been found that many.Skeletal muscle efficiency operation relates to energy matter picked-up and metabolism, myogen kind and quantity, calcium ion Rhizoma Nelumbinis connection and the many factors such as concentration, wherein, improve energy supply and picked-up be the most directly, the most controlled and most efficient method.

Glucose transporter 4 (Glucose transporter type 4 on skeletal muscle cell membrane, GLUT4) being that glucose molecule enters intracellular unique passage from skeletal muscle cell membrane, namely there is the main transporter that easyization spreads in glucose on cell membrane.It plays the effect of Skeletal Muscle Cell energy feedstuff supply pump.Its gene expression dose and film indexing decide the ingestion efficiency of glucose.GLUT4 is by 509 Amino acid profiles, and relative molecular mass is 45 000-55 000, is primarily present in skeletal muscle, cardiac muscle and fatty tissue.Under rest state, the GLUT4 overwhelming majority is distributed on intracellular vesicle film, and when insulin secretion or Motor stimulation, GLUT4 is indexed into cell membrane and structural change occurs, and carries out glucose uptake, provides energy matter for skeletal muscle activity.In body, the glucose of 85% is to directly feed skeletal muscle picked-up and utilize.Therefore, promote gene expression and the film indexing of GLUT4 in Skeletal Muscle Cell, be to increase skeletal muscle glucose supply, improve motor capacity important method and approach.

Radix Ginseng is described as " king of BAICAO " in China, is one of Relict Plant the most ancient on the earth.The most on the books at multi-section Chinese materia medica monographs such as Shennong's Herbals.Containing multiple ginsenoside and polysaccharide bioactive compound in Radix Ginseng.There is anticancer, blood sugar lowering, radioprotective, growth promotion, increasing vigor and anti-ageing function of waiting for a long time.

Ginsenoside Rg1 is one of most active bioactive ingredients in Radix Ginseng, and its pharmacological action is had made extensive and intensive studies by we.Result of study shows: ginsenoside Rg1 can promote gene expression and the film indexing of GLUT4 in Skeletal Muscle Cell, it is possible to strengthens the supply of Skeletal Muscle Cell energy, improves body movement ability.

Summary of the invention

It is an object of the invention to provide a kind of ginsenoside Rg1 and be applied to the health product of preparation raising body movement ability.

Experimental result shows, glucose uptake and utilization are had a very important role by the gene expression of GLUT4 and film indexing.Ginsenoside Rg1 can strengthen gene expression and the film indexing of GLUT4, thus promotes that Skeletal Muscle Cell is to glucose uptake and utilization.

First laboratory cultures sarcoplast C2C12 (being purchased from Rui Chu bio tech ltd, Shanghai)；Three kinds of variable concentrations (10 on probation, 20,40 μMs) Rg1 (being purchased from prophyll bio tech ltd, Shanghai, Nanjing, purity is 98.0%) hatch altogether with C2C12, GLUT4 is expressed by 20 μMs of concentrations above ginsenoside Rg1s significant facilitation.The experimental techniques such as application RT-PCR, Westernblot, it was demonstrated that after ginsenoside Rg1 and C2C12 cell are hatched altogether, detecting that in C2C12 cell, GLUT4 genetic transcription increases, GLUT4 protein level raises, and GLUT4 is strengthened to after birth indexing by endochylema.

Beneficial effect: the essence improving body movement ability is to strengthen skeletal muscle acting, increasing the supply of Skeletal Muscle Cell energy matter is to strengthen skeletal muscle acting important channel.Ginsenoside Rg1 can promote the expression of Skeletal Muscle Cell GLUT4 and film indexing, improves skeletal muscle to the picked-up of glucose sugar and utilization, and direct support bones flesh does work, and improves body movement ability.

Accompanying drawing explanation

Fig. 1 is that after ginsenoside Rg1 is hatched altogether with C2C12 cell, RT-PCR detects GLUT4mRNA transcriptional level.

Fig. 2 is that after ginsenoside Rg1 is hatched altogether with C2C12 cell, Western blot detects GLUT4 protein expression level.

Fig. 3 is that after ginsenoside Rg1 is hatched altogether with C2C12 cell, Western blot detects cell membrane GLUT4 protein level.

Fig. 4 is after ginsenoside Rg1 is hatched altogether with C2C12 cell, and glucose oxidizing process detection glucose uptake utilizes level.

Detailed description of the invention

1.C2C12 cell is cultivated

Taking out frozen C2C12 cell from-80 DEG C of refrigerators, be quickly transferred in 37 DEG C of water-baths make cell instant, 800r is centrifuged 5 minutes, abandons supernatant.With 10% hyclone DMEM culture fluid at 37 DEG C, 5%CO₂In cell culture incubator.Change low concentration serum free culture system liquid and cultivate its differentiation of rush.Change DMEM height sugar culture fluid to cultivate, culture fluid adds ginsenoside Rg1 and hatches altogether.

2.RT-PCR detects GLUT4mRNA

C2C12 cell after incubation 24h, is used Trizol to extract total serum IgE in C2C12 cell, is detected by OD value at 260nm and 280nm in containing ginsenoside Rg1's culture fluid, measures extracted RNA amount and purity.The RT-PCR kit explanation provided according to Takara company carries out reverse transcription and amplification experiment, and primer sequence is as follows: GLUT4 forward primer is 5 '-AGGCTGCCTATAAACACTGG-3 '；GLUT4 downstream primer is 5 '-AGGAAGGATGAAGTGCAAAG-3 '.Selecting annealing temperature is 60 DEG C.1.5% agarose gel electrophoresis is used to analyze PCR primer amount.

3.Western blot detection GLUT4 expresses

C2C12 cell after incubation 24h, extracts total protein and carries out SDS-PAGE in containing ginsenoside Rg1's culture fluid.With Semi-Dry electroporation by the Protein transfer in gel to pvdf membrane.Pvdf membrane after electricity turns is dehydrated in methanol, and defatted milk powder is closed, and an antibody (mountain sheep anti mouse) is hatched jointly with pvdf membrane, and TBST washs, and two antibody (rabbit anti goat igg) are hatched, and TBST washs, with ECL Kit testing goal albumen.

4.Western blot detection GLUT4 film indexing

C2C12 cell is in containing ginsenoside Rg1's culture fluid after incubation 24h, and cell separates through cold phosphate buffer eluting, homogenate, differential centrifugation, plasma membrane, extracts memebrane protein and carries out SDS-PAGE.Method is with 3.

5. glucose consumption detection

C2C12 cell in containing ginsenoside Rg1's culture fluid after incubation 24h, the glucose content in detection culture fluid, with the sugared content average in the multiple hole of non-inoculating cell blank for comparison, calculate other each multiple porocyte consumption to glucose.

6. interpretation of result

(1) ginsenoside Rg1 promotes that C2C12GLUT4mRNA transcribes

The impact that C2C12 cell GLUT4mRNA is transcribed by RT-PCR experiment detection ginsenoside Rg1.RT-PCR result shows, C2C12 cell GLUT4mRNA has been transcribed the effect of being obviously promoted by ginsenoside Rg1, and has concentration dependent, such as Fig. 1.In figure, 1 is matched group, is not added with ginsenoside Rg1；2-4 is that ginsenoside Rg1 hatches group；Ginsenoside Rg1's concentration is respectively 10,20 and 40uM.Wherein, 20 and 40uM groups remarkably promote effect to GLUT4 cell mRNA transcriptional level.

(2) ginsenoside Rg1 promotes C2C12 cell GLUT4 protein expression

Western blot experiment detection ginsenoside Rg1's impact on C2C12 cell GLUT4 protein expression.Western blot result shows, ginsenoside Rg1 is obviously promoted effect to C2C12 cell GLUT4 protein expression, and has concentration dependent, such as Fig. 2.In figure, 1 is matched group, is not added with ginsenoside Rg1；2-4 is that ginsenoside Rg1 hatches group；Ginsenoside Rg1's concentration is respectively 10,20 and 40uM.Wherein, 20 and 40uM groups remarkably promote effect to GLUT4 protein expression.

(3) ginsenoside Rg1 promotes the indexing of C2C12 cell GLUT4 film

The Western blot experiment detection ginsenoside Rg1 impact on the indexing of C2C12 cell GLUT4 albuminous coat.Western blot result shows, ginsenoside Rg1 is obviously promoted effect, such as Fig. 3 to C2C12 cell GLUT4 albuminous coat indexing.In figure, ginsenoside Rg1 hatches group (20 and 40uM) C2C12 cell membrane GLUT4 protein level has notable rising than matched group.

(4) ginsenoside Rg1 promotes C2C12 grape cell Sugar intake

Glucose oxidizing process experiment detection ginsenoside Rg1 utilizes impact on C2C12 cell to glucose uptake.Glucose oxidation experiment result shows, ginsenoside Rg1 can promote C2C12 grape cell Sugar intake, such as Fig. 4.During in figure, ginsenoside Rg1 hatches group (40uM) cell culture fluid, glucose level significantly reduces.

In summary it can be seen, ginsenoside Rg1 can promote osteoblast C2C12GLUT4 genetic transcription and protein expression, and can promote the indexing of GLUT4 film, strengthens C2C12 to the picked-up of glucose and utilization, there is enhancing skeletal muscle acting, improve the effect of body movement ability.

Claims

1. ginsenoside Rg1's application in preparation improves body movement ability health product.It is characterized by: ginsenoside Rg1 can promote Enter the genetic transcription of GLUT4, improve protein expression level and film indexing, thus strengthen Skeletal Muscle Cell to the picked-up of glucose and Utilize.