CN109294980A - Root of kirilow rhodiola and rhodioside are divided into the application in cardiac-like muscle cell in stem cell directional - Google Patents
Root of kirilow rhodiola and rhodioside are divided into the application in cardiac-like muscle cell in stem cell directional Download PDFInfo
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Abstract
It is divided into the application in cardiac-like muscle cell, especially rhodioside in induction stem cell directional the present invention provides root of kirilow rhodiola and rhodioside and combine with hypoxemia and be used for induced lipolysis mescenchymal stem cell directed differentiation as inducer as cardiac-like muscle cell.The present invention has opened up the new application of rhodioside, i.e., the stem cell directionals such as external evoked fat stem cell are divided into cardiac-like muscle cell；The effect that root of kirilow rhodiola and rhodioside are divided into cardiac-like muscle cell in inducing adult stem cell directional is provided, is conducive to internal cellular replacement therapy acute and chronic myocardial damage and the further investigation in cardiovascular disease field, provides the foundation for new drug development.
Description
Technical field
The present invention relates to root of kirilow rhodiola especially rhodiosides to be divided into answering in cardiac-like muscle cell in induction stem cell directional
With.
Background technique
Root of kirilow rhodiola system red-spotted stonecrop (Crassulaceae) section rhodiola (Rhodiola L.) plant, be perennial herb or
Semishrub plant is mainly distributed on the Himalayas, the Asia northwestward and North America height above sea level in the Northern Hemisphere 1600~4000
The area of rice.Root of kirilow rhodiola medicinal history is long, is known as the title of " gold ginseng ".It is crassulaceae plants great Hua that pharmacopeia, which records root of kirilow rhodiola,
The dry root and stem root of root of kirilow rhodiola, property is sweet, bitter, flat, qi and activate blood circulation, promotes blood circulation and relievings asthma, can be used for qi deficiency to blood stasis, chest impediment and cardialgia,
Hemiplegia and burnout asthma.Modern pharmacological studies have shown that the bioactivity of root of kirilow rhodiola and a variety of chemical components contained by it
It is inseparable.
Rhodioside (salidroside) is the natural products of the separating-purifying from root of kirilow rhodiola, is a kind of monomeric compound,
It is one of the effective component of root of kirilow rhodiola, the related section in part belongs to Chinese medicine and also contains rhodioside, and pharmacological action has following several respects:
It improves and improvement resists various altitude sickness prevention abilities.
Anti anoxia, antifatigue effect.Research shows that it is by maintaining 5-HT content normalization to play anti-central nerve fatigue
Effect.
Anti-aging, with antioxidation and the external ability for understanding free radical and ultra-oxygen anion free radical.
Inhibit rat renal tubular epithelial cells and interstitial cell to convert to myofibroblast, renal interstitial damage is effectively relieved
Wound, prevents kidney region fibrosis.
Anti-fibrosisization effect.The HSC increment and the expression of glue protogene that rhodioside can obviously inhibit acetaldehyde to stimulate,
There is significant effect of anti hepatic fibrosis in vitro.
Protective effect to cardiovascular and cerebrovascular.Rhodioside can inhibit interleukins I β table caused by Ischemia and Reperfusion in vivo in Rats
The increase reached, the protection played the role of of expression for reducing TNF-α in cerebral ischemia re-pouring tissues following MCAO in rats is cerebrovascular, passes through inhibition
The expression of blood vessel ACE can resist myocardial ischemia to a certain extent.
Improve body immunity, antitumor, antiviral.
It is anti-radiation.Rhodioside plays the role of protecting X-ray damage body tissue and cell membrane.
It improves memory and prevents senile dementia, protect nerve cell.Rhodioside can inhibit caused by oxygen-glucose deprived injury
Mitochondrial membrane potential and active reduction to have the function of stablizing mitochondrial membrane potential inhibit nerve cell apoptosis
Occur；The external clinic for promoting neural stem cell to think that neuron direction is divided into root of kirilow rhodiola drug for related neurological disease
Treatment provides experimental basis.
Stem cell can be divided into function under certain condition as a kind of cell with self-renewing and Multidirectional Differentiation
The islet cells of energy property, therefore can be used as the completely new source of islet cells.Breaking up acquisition cardiac-like muscle cell by stem cell at present has
Several approach: ES cell differentiation and adult stem cells.It is thin into soma since embryonic stem cell has dispute of ethic
Born of the same parents are from a wealth of sources, and no dispute of ethic can be used as the source of cardiac-like muscle cell.
Contain a kind of cell with Multidirectional Differentiation potentiality, i.e. fat mesenchymal stem cell, referred to as fat in adipose tissue
Stem cell.Its biological property is similar with mesenchymal stem cell, and can to fat, bone, cartilage, muscle, endothelium, liver,
The differentiation of the various kinds of cell such as pancreas islet and nerve direction.Due to adipose tissue in human body rich reserves, it is small to obtain easy wound, in group
Weaver's journey, organ reparation, gene therapy etc. suffer from wide application prospect, therefore fat mesenchymal stem cell has become
Another hot spot for being concerned of stem cell field after mesenchymal stem cell.
Current stem cell induction mode mainly has external evoked, gene modification, protein transduction and tissue microenvironment to induce,
Wherein external evoked is that stem cell is induced to differentiate into aim cell using different stimulated combinations of factors.Each laboratory-induced point
Change condition is different, and induction differentiation mechanism is still not clear, and induction differentiation efficiency is low, and islet secretion ability is only that normal islets 1% are left
The right side, and Induction Process is complicated, induction time is long, and gained cell quantity is few, and function is low.
There are not also root of kirilow rhodiola and rhodioside that there is induction stem cell directional to be divided into cardiac-like muscle cell and repair the heart at present
The effect of injury of muscle is reported.
Summary of the invention
The purpose of the present invention is to provide root of kirilow rhodiola and the new application of rhodioside, i.e. root of kirilow rhodiola and rhodioside induction is dry
Cell directional is divided into the application of cardiac-like muscle cell.
To achieve the above object, the technical solution adopted by the present invention are as follows:
Root of kirilow rhodiola or rhodioside are divided into the application in cardiac-like muscle cell inducer in preparation induction stem cell directional.
Preferably, inducer induces stem cell directional to be divided into cardiac-like muscle cell under low oxygen conditions.
Preferably, the hypoxia condition refers to that the volumn concentration of oxygen is 2~14%.
Preferably, the stem cell is fat mesenchymal stem cell.
The present invention also provides the inducers that a kind of induction stem cell directional is divided into cardiac-like muscle cell, which is characterized in that institute
It states and contains rhodioside in inducer.
Preferably, the inducer induces stem cell directional to be divided into cardiac-like muscle cell by induced medium, described red
Concentration of the red-spotted stonecrop glycosides in the induced medium is 1~400 μM.
The present invention also provides the abductive approach that a kind of induction stem cell directional is divided into cardiac-like muscle cell, which is characterized in that
The following steps are included:
It takes animal tallow mescenchymal stem cell to carry out secondary culture, selects the fat mesenchymal stem cell for stablizing passage low
It is incubated in the induced medium containing rhodioside and is induced under the conditions of oxygen, differentiation obtains Mature myocardium like cell.
The present invention also provides rhodiosides in the drug for being prepared by stem-cell therapy myocardial infarction, myocarditis, coronary heart disease
In application.
Preferably, when the drug is oral preparation, mass percentage of the rhodioside in the drug is big
In 20%；When the drug is ejection preparation, mass percentage of the rhodioside in the drug is greater than 5%.
Preferably, when the drug is oral preparation, mass percentage of the rhodioside in the drug is big
In 10%；When the drug is ejection preparation, mass percentage of the rhodioside in the drug is greater than 2%.
Preferably, the oral preparation is tablet, capsule, particle, dripping pill, oral solution or outstanding mixture.
It is an object of the invention to also provide a kind of drug using stem-cell therapy myocardial damage related disease, the medicine
Contain rhodioside in object；Preferably, when the drug is oral preparation, quality hundred of the rhodioside in the drug
Content is divided to be greater than 20%；When the drug is ejection preparation, mass percentage of the rhodioside in the drug is big
In 5%.
Preferably, when the drug is oral preparation, mass percentage of the rhodioside in the drug is big
In 10%；When the drug is ejection preparation, mass percentage of the rhodioside in the drug is greater than 2%.
The beneficial effects of the present invention are: the present invention provides root of kirilow rhodiola and rhodioside in induction stem cell directional differentiation
Combine for the application in cardiac-like muscle cell, especially rhodioside with hypoxemia and is done carefully as inducer for induced lipolysis mesenchyma
Born of the same parents' directed differentiation is cardiac-like muscle cell.The present invention has opened up the new application of rhodioside, i.e., external evoked fat stem cell etc. is dry
Cell directional is divided into cardiac-like muscle cell；It provides root of kirilow rhodiola and rhodioside and is divided into cardiac muscle in inducing adult stem cell directional
Effect in like cell is conducive to deeply grinding for internal cellular replacement therapy acute and chronic myocardial damage and cardiovascular disease field
Study carefully, provides the foundation for new drug development.
Detailed description of the invention
Fig. 1 be fat mesenchymal stem cell growth conditions schematic diagram, wherein A be the 3rd fat subsitutes mescenchymal stem cell (×
100), the 3rd generation cell (× 100) of B recovery.
Fig. 2 is the qualification result schematic diagram of fat mesenchymal stem cell transdifferentiation cardiac-like muscle cell, and wherein A is cTnI albumen
Express (take on a red color fluorescence, positive cell, × 400), B be connexin43 protein expression (be in green fluorescence, positive cell, ×
400)。
Fig. 3 is that rhodioside and hypoxemia induce transdifferentiation cardiac-like muscle cell correlating markings albumen to fat mesenchymal stem cell
The schematic diagram of (cTnI, cTnT, α-actinin and GATA-4) expression.
Fig. 4 is that rhodioside and hypoxemia induce transdifferentiation cardiac-like muscle cell correlating markings albumen to fat mesenchymal stem cell
The significance difference analysis schematic diagram of (cTnI, cTnT, α-actinin and GATA-4) expression.(* indicates P < 0.5, * * table
Show that P < 0.01, * * * indicate P < 0.001.)
Fig. 5 is that rhodioside and hypoxemia induce transdifferentiation cardiac-like muscle cell related regulatory factors to fat mesenchymal stem cell
(Notch1, Jagged1 and Hes1) expression schematic diagram.
Fig. 6 is that rhodioside and hypoxemia induce transdifferentiation cardiac-like muscle cell related regulatory factors to fat mesenchymal stem cell
The significance difference analysis schematic diagram of (Notch1, Jagged1 and Hes1) expression.(* expression P < 0.5, * * expression P <
0.01, * * * indicates P < 0.001.)
Specific embodiment
For more concise displaying technical solution of the present invention, objects and advantages, combined with specific embodiments below
And attached drawing is described in further detail the present invention.
In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition, such as " molecular cloning "
Condition described in (New York:Cold Spring Harbor Laboratory Press, 1989), or according to manufactory
Condition proposed by quotient
Embodiment 1
Reagent and material
Rhodioside (Sigma Co., USA), fat mesenchymal stem cell (ASCs) (Sai Ye Biotechnology Co., Ltd),
The complete DMEM of fat mesenchymal stem cell (Sai Ye Biotechnology Co., Ltd), Osteoinductive differentiation culture medium (match industry biology section
Skill Co., Ltd), adipogenic induction differential medium (Sai Ye Biotechnology Co., Ltd), (U.S. Gibco is public for DMEM culture medium
Department), dual anti-mixed liquor (Beijing Suo Laibao company), fetal calf serum (Hyclone company of the U.S.), rabbit-anti mouse cTnT polyclonal antibody
(Cell Signaling Technology company of the U.S.), rabbit-anti mouse cTnI polyclonal antibody (U.S. Cell Signaling
Technology company), rabbit-anti mouse α-actinin polyclonal antibody (Cell Signaling Technology company of the U.S.),
Rabbit-anti mouse GATA-4 polyclonal antibody (Cell Signaling Technology company of the U.S.).
Experimental procedure:
(1) fat mesenchymal stem cell culture
Rat adipose-derived mesenchymal stem cells are purchased from Biosciences Inc. (Guangzhou, China).It is grafted respectively in 75cm2Training
Bottle is supported, basal medium, 10% fetal calf serum of addition, 2mM glutamine and 1% Penicillin Streptomycin Solution are incubated at.It will be thin
Born of the same parents cultivate in 5%CO2,37℃.Every two days one subcultures of replacement pass on (as shown in Figure 1A) when about 80% converges.
(2) root of kirilow rhodiola induced lipolysis mescenchymal stem cell directed differentiation is cardiac-like muscle cell
The ASCs for taking culture 3 generations of ground is inoculated in Tissue Culture Dish (containing 2% fetal calf serum, 100U/ml penicillin, 100U/
The low sugar DMEM culture medium of ml streptomysin) 37 DEG C, 5%CO2Under the conditions of culture to cell confluency rate be greater than 90% (such as Figure 1B institute
Show), cell culture medium is changed to basal medium, is divided into 3 experimental groups (Normoxia+SAL, NS group) and 1 control group
(NC group) is added respectively in basal medium:
Control group: basis culture
Rhodioside induction group (NS group): it is respectively 1 μM, 200 μM, 400 μ that root of kirilow rhodiola induction group, which is equipped with 3 various concentrations,
M；
It is primary to change within every 3 days liquid 1, and observes the variation of cellular morphology, the 5 days harvest cells in culture ground, by detection cTnI and
Connexin43 albumen identifies that fat mesenchymal stem cell is divided into cardiac muscle cell's situation, passes through Western Blot and detects rouge
Fat mescenchymal stem cell Myocardial marker cTnI and, the expression of cTnT, α-actinin and GATA-4.
Embodiment 2
The reagent and material of the present embodiment are same as Example 1.
Experimental procedure:
(1) fat mesenchymal stem cell culture: the present embodiment step (1) is same as Example 1.
(2) hypoxemia processing induced lipolysis mescenchymal stem cell directed differentiation is cardiac-like muscle cell
The ASCs for taking culture 3 generations of ground is inoculated in Tissue Culture Dish (containing 2% fetal calf serum, 100U/ml penicillin, 100U/
The low sugar DMEM culture medium of ml streptomysin) 37 DEG C, 5%CO2Under the conditions of culture to cell confluency rate be greater than 90%, by cell culture
Base is changed to basal medium, and 3 experimental groups are arranged and cultivate under different hypoxia conditions respectively.
Hypoxia inducible group (Hypoxia, HC group): the oxygen content of three not isogeneous induction groups is respectively 2%, 5%, 14%；
It is primary to change within every 3 days liquid 1, and observes the variation of cellular morphology, the 5 days harvest cells in culture ground pass through Western
The expression of Blot detection fat mesenchymal stem cell Myocardial marker cTnI, cTnT, α-actinin and GATA-4.
Embodiment 3
The reagent and material of the present embodiment are same as Example 1.
Experimental procedure:
(1) fat mesenchymal stem cell culture: the present embodiment step (1) is same as Example 1.
(2) rhodioside and hypoxemia Combined Treatment induced lipolysis mescenchymal stem cell directed differentiation are cardiac-like muscle cell
The ASCs for taking culture 3 generations of ground is inoculated in Tissue Culture Dish (containing 2% fetal calf serum, 100U/ml penicillin, 100U/
The low sugar DMEM culture medium of ml streptomysin) 37 DEG C, 5%CO2Under the conditions of culture to cell confluency rate be greater than 90%, by cell culture
Base is changed to basal medium, is divided into 3 experimental groups, and grouping and experiment condition are as follows:
Rhodioside+hypoxemia processing induction group (Hypoxia+SAL, HS group): the induction group sets that there are three different root of kirilow rhodiola
Glycosides concentration is combined with low oxygen content, is respectively 1 μM of+2%O2, 200 μM of+5%O2, 400 μM of+14%O2；
It is primary to change within every 3 days liquid 1, and observes the variation of cellular morphology, the 5 days harvest cells in culture ground pass through Western
The expression of Blot detection fat mesenchymal stem cell Myocardial marker cTnI, cTnT, α-actinin and GATA-4.
By the specific proteins of above-described embodiment 1-3 cardiac-like muscle cell extracted (cTnI, cTnT, α-actinin and
GATA-4 the case where expression) is detected, embodiment 1-3 each group is distributed, and is shown in Table 1.
Table 1: the case where embodiment 1-3 each group, is distributed
Laboratory test results:
From Figure 2 it can be seen that cTnT albumen (red fluorescence, positive cell, × 400) and connexin43 albumen (green fluorescence,
Positive cell, × 400) it has been expressed, illustrate that fat mesenchymal stem cell is successfully divided into cardiac-like muscle cell.
As seen from Figure 3, compared with NC group, the expression quantity of cTnT and GATA-4 in NS group are above NC group, in HC group
GATA-4 expression quantity is higher, and the expression quantity of cTnI, cTnT, α-actinin and GATA-4 of HS group are above HC group.Meanwhile
In NS group, the expression quantity highest of MGATA-4 when rhodioside concentration is 400 μ；In HC group, α-when low oxygen content is 5%
The expression quantity highest of actinin；In HS group, the concentration of rhodioside is 200 μM, cTnI, α-when low oxygen content is 5%
The equal highest of the expression quantity of actinin and GATA-4, therefore, the concentration of rhodioside are 200 μM, are this when low oxygen content is 5%
The most preferred embodiment of invention.
As shown in Figure 4, compared to control group (NC group), cTnI is increased to 2.4 times of NC group, cTnT liter in NS group in HC group
Up to 1.4 times of NC group have statistical difference, illustrate that rhodioside and hypoxemia may act on different troponins；And HS
CTnI and cTnT is increased to 3.5 and 1.7 times of NC group respectively in group, and has significant difference, illustrates rhodioside and hypoxemia
Combined pretreatment has facilitation for two kinds of troponins, is better than simple rhodioside or hypoxic preconditioning (Fig. 4 A, B).
Likewise, rhodioside joint hypoxic preconditioning can promote the expression of α-actinin and GATA-4, and it is better than simple root of kirilow rhodiola
Glycosides or hypoxic preconditioning (Fig. 4 C, D).
The molecule of 4 rhodioside of embodiment and the differentiation of hypoxic preconditioning induced lipolysis mescenchymal stem cell Cardiocytes
Mechanism
One, experimental material
Rhodioside (Sigma Co., USA), DMEM culture medium (Sai Ye Biotechnology Co., Ltd), dual anti-mixed liquor
(Beijing Suo Laibao company), fetal calf serum (Hyclone company of the U.S.), BCA measure protein concentration reagent (the green skies biology in Shanghai
Technology Co., Ltd.), rabbit-anti mouse Akt polyclonal antibody (Cell Signaling Technology company of the U.S.), rabbit-anti mouse p-
Akt polyclonal antibody (Cell Signaling Technology company of the U.S.), rabbit-anti mouse Erk polyclonal antibody (U.S. Cell
Signaling Technology company), rabbit-anti mouse p-Erk polyclonal antibody (U.S. Cell Signaling Technology
Company).
Two, experimental method
1, experimental group:
The ASCs for taking culture 3 generations of ground is inoculated in Tissue Culture Dish (containing 2% fetal calf serum, 100U/ml penicillin, 100U/
The low sugar DMEM culture medium of ml streptomysin) 37 DEG C, 5%CO2Under the conditions of culture to cell confluency rate be greater than 90%, by cell culture
Base is changed to basal medium, 1 control group (NC group) and 3 experimental groups, respectively rhodioside group (NS group), hypoxemia processing
Group (HC group), rhodioside and hypoxemia Combined Treatment group (HS group), every group three are parallel:
Control group: basis culture
Rhodioside induction group (NS group): rhodioside concentration is 50 μ Μ；
Hypoxemia processing group (HC group, oxygen content 5%O2)；
(HS group, rhodioside concentration are 50 μM of+5%O for rhodioside and hypoxemia Combined Treatment group2)；
It is primary to change within every 3 days liquid 1, and observes the variation of cellular morphology, the 5 days harvest cells in culture ground, by detection cTnI and
Connexin43 albumen identifies that fat mesenchymal stem cell is divided into cardiac muscle cell's situation, passes through Western Blot and detects rouge
Fat mescenchymal stem cell Myocardial marker cTnI and, the expression of cTnT, α-actinin and GATA-4.
2, experimental method
(1) by the cell protein of extraction, by BCA method protein solution concentration mensuration, carry out transferring film after electrophoresis (90V,
120min)；Prepare TBS (1 packet powder+2L distilled water) → TBS-T (500ml TBS+500 μ l Tween20)
(2) enzyme immunolocalization, closing: the pvdf membrane after transferring film is washed 5 times, each 5min with TBS-T, with containing 5% degreasing
Milk powder confining liquid room temperature closes 2h.It incubates primary antibody: being washed film 5 times, each 5min after closing with TBS-T, is incubated for primary antibody confining liquid, 4
DEG C overnight.Incubate secondary antibody: recycling primary antibody confining liquid, TBS-T are washed film 5 times, each 5min, are incubated for 1h, recycling two with secondary antibody diluent
Anti- dilution, TBS-T washing.
(3) it develops the color, expose: pvdf membrane being reacted into 5min in ECL chemiluminescent agent, dries, is placed in X-ray magazine, film
5min is exposed, film development, fixing are taken out.
(4) it gray analysis: after film scanning, is analyzed in Image J image analysis software.
Three, experimental result
Notch signal path plays a significant role during heart development, by detecting Notch signal path receptor egg
The expression of white Notch1 and its ligandin Jagged1 and target protein Hes1, with clear Notch1 access between fat
The effect of mesenchymal stem cells.As a result, it has been found that (as shown in Figure 5,6), compared to NC group, Jagged1 is increased to NC group in NS group
3.1 times and there is statistical difference, but Jagged1 is horizontal in HC group and indifference；And Jagged1 is increased to NC group in HS group
7.5 times, and all have significant difference between HS group and other each groups, illustrate that rhodioside joint hypoxic preconditioning can be significant
Promote the synthesis of ligandin Jagged1.In addition, the expression of Notch1 rises in NS group, HC group and HS group compared to NC group
Height all has statistical difference, illustrate each pretreated group can upregulated receptor albumen Notch1 level.Next we send out
Existing, compared to NC group, NS group and HC group, the expression of Hes1 is lowered in HS group, and all has statistical difference, illustrates rhodioside
Joint hypoxic preconditioning group can lower the level of target protein Hes1, so that ASCs Cardiocytes be promoted to break up.
Four, experiment conclusion
The synthesis for the active ligand Jagged1 that rhodioside joint hypoxic preconditioning remarkably promotes Notch1 first is (significant
Better than the effect of simple rhodioside and hypoxic preconditioning) so that receptor protein Notch1 raise, ligandin Jagged1 and
After receptor protein Notch1 is combined, by intracellular cascade reaction, acts on target protein Hes1 and lower its expression, thus
Play the effect for promoting the differentiation of ASCs Cardiocytes.
Embodiment 5
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into tablet, takes orally, wherein
Determination of Salidroside 0.2g in per daily amount of formulation.
Embodiment 6
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into capsule, takes orally,
In per Determination of Salidroside 0.2g in daily amount of formulation.
Embodiment 7
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into granule, takes orally,
In per Determination of Salidroside 0.2g in daily amount of formulation.
Embodiment 8
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into pill, takes orally,
In per Determination of Salidroside 0.2g in daily amount of formulation.
Embodiment 9
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into oral solution, mouth
Clothes, wherein Determination of Salidroside 0.2g in per daily amount of formulation.
Embodiment 10
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into outstanding mixture, takes orally,
In per Determination of Salidroside 0.2g in daily amount of formulation.
Embodiment 11
It takes rhodioside as bulk pharmaceutical chemicals, medically acceptable auxiliary material composition is added, is prepared into ejection preparation, takes orally,
Wherein Determination of Salidroside 0.05g in every daily amount of formulation.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. root of kirilow rhodiola or rhodioside are divided into the application in cardiac-like muscle cell inducer in preparation induction stem cell directional.
2. application as described in claim 1, which is characterized in that inducer induces stem cell directional to be divided under low oxygen conditions
Cardiac-like muscle cell.
3. application as claimed in claim 2, which is characterized in that the hypoxia condition refers to that the volumn concentration of oxygen is 2
~14%.
4. application as described in claim 1, which is characterized in that the stem cell is fat mesenchymal stem cell.
5. the inducer that a kind of induction stem cell directional is divided into cardiac-like muscle cell, which is characterized in that contain in the inducer
Rhodioside.
6. inducer as claimed in claim 5, which is characterized in that the inducer induces stem cell fixed by induced medium
To cardiac-like muscle cell is divided into, concentration of the rhodioside in the induced medium is 1~400 μM.
7. the abductive approach that a kind of induction stem cell directional is divided into cardiac-like muscle cell, which comprises the following steps:
It takes animal tallow mescenchymal stem cell to carry out secondary culture, selects the fat mesenchymal stem cell for stablizing passage in hypoxemia item
It is incubated in the induced medium containing rhodioside and is induced under part, differentiation obtains Mature myocardium like cell.
8. application of the rhodioside in the drug for being prepared by stem-cell therapy myocardial infarction, myocarditis or coronary heart disease.
9. application as claimed in claim 8, which is characterized in that when the drug is oral preparation, the rhodioside is in institute
The mass percentage stated in drug is greater than 20%；When the drug is ejection preparation, the rhodioside is in the drug
Mass percentage be greater than 5%.
10. a kind of drug using stem-cell therapy myocardial damage related disease, which is characterized in that contain red scape in the drug
Its glycosides；Preferably, when the drug is oral preparation, mass percentage of the rhodioside in the drug is greater than
20%；When the drug is ejection preparation, mass percentage of the rhodioside in the drug is greater than 5%.
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