CN105713972A - Application of miRNA to preparation of drug-induced heart disease biomarkers - Google Patents
Application of miRNA to preparation of drug-induced heart disease biomarkers Download PDFInfo
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Abstract
The invention belongs to the field of biotechnology and particularly relates to application of miRNA to preparation of drug-induced heart disease biomarkers.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of miRNA (microRNA) purposes in preparing drug-induced heart disease biomarker.
Background technology
Broad sense and discuss; the heart change that all myocardial toxicity direct or indirect due to medicine cause; no matter it is the exception of the myocardium depolarization because the impact of cardiac electrophysiology is caused by medicine and multipole or various arrhythmia; or the contractility of cardiac muscle is suppressed because of the toxic action of medicine; and then the heart failure bringing out or increasing the weight of; or because certain drug allergy is caused myocarditis, pericarditis, drug-induced heart disease (drug-inducedheartdiseases) all can be referred to as.The same with other drug-induced disease, drug-induced heart disease can be facilitated due to the reason of medicine itself, the doctor of medication and patient three aspect controlled, but generally medicine itself and drug interaction are the leading factor causing pathological changes.Medicine involves heart, and person is mainly the infringement to cardiac muscle, but minority medicine also involves pericardium and endocardium.
Radix Tripterygii Wilfordii (TripterygiumwilfrdiiHook.f.) has another name called Caulis Fibraureae, Caulis Fibraureae wood, allusion quotation wax rattan, Gelsemium elegans Benth. etc., belongs to Wei Mao section bejuco.Its English name: CommonThreewingnutRoot, latin name: RadixTripterygiiWilfordiiTripterygiumwilfordiiHooK.f.Documents and materials record its bitter in the mouth, pungent, cool in nature, poison, returns liver, kidney channel greatly.There is the effects such as expelling wind and removing dampness, promoting blood circulation to remove obstruction in the collateral, reducing swelling and alleviating pain, parasite killing removing toxic substances.The root of Radix Tripterygii Wilfordii, neck, Ye Junke are as medicinal, and main medicinal part is its root.Finding in clinical practice, the mechanism of Tripterygium Preparations treatment rheumatoid arthritis is similar to corticosteroids, but does not have the side effect of hormones, and better tolerance, potential applicability in clinical practice is wide.But research in recent years finds, Radix Tripterygii Wilfordii may result in toxic myocarditis, causes cardiac conduction obstacle, and severe patient may occur in which cardiogenic shock and heart failure.In the acute toxic cardiac damage that various Tripterygium Preparations cause, the cardiac damage caused with triptolide again is the most serious.
Biomarker (Biomarker) refer to can Mk system, organ, tissue, the changing or the biochemical indicator of contingent change of cell and subcellular structure or function, there is purposes widely.Biomarker can be used for medical diagnosis on disease, judges staging or for evaluating new drug or the new therapy safety and efficacy in target group.The biochemical indicator of diagnosing cardiac has been widely used in the detection of cardiotoxicity of medicine clinically at present.Wherein, cardiac muscle troponin I (cTnI) is specific to a kind of myocardial cell, exists only in the albumen in ventricular muscles and heart muscle.The molecular weight of cTnI is 22ku, and serum half-life is about 2h.CTnI there are about 3%～6% in cardiac muscle and is present in a free form in endochylema, and remainder is then present in sarcostyle with combining form.In normal human serum, the content of cTnI is little, and after acute myocardial infarction (AMI) is fallen ill, serum cTnI starts to raise more than 3～6h, and 10～24h reaches peak, and 5～7d recovers normal.Compared with cardiac muscle creatine kinase mb (CK-MB), cTnI content in cardiac muscle is more, and molecular weight is less than normal, easily measured in serum after myocardial damage, so it is comparatively sensitive, can be used for the microlesion of detection cardiac muscle.
MiRNA (microRNA) is a newfound non-coding strand microRNA of class, about 22 length of nucleotides, by identifying specific target mRNA, with the 3 ' of said target mrna untranslated region (3 '-untranslatedregion, 3 '-UTR) complementary pairing, carry out protein translation suppression or degraded mRNA.The growth promoter of miRNA wide participation body, the metabolism of cell, propagation, differentiation and apoptosis, take part in the multiple pathological processes such as hepatic injury, tumor formation, cardiovascular disease.Therefore, in normal body, or even the change of miRNA also indicates that the change of body the normal physiological function generation of pathological changes.Existing more than 700 kind of miRNA is found, and records in miRBase data base, and current miRNA has become the focus of life sciences circle research.The various disease conditions such as tumor, cardiovascular and cerebrovascular disease has been carried out by miRNA as the research for the purpose of diagnosis index and therapy target by existing research worker, and has made some progress.Owing to the miRNA in the biological specimen such as blood plasma, tissue remains to detect after room temperature, high temperature, different pH, thawing, and changes of contents is little, and now most of RNA, protein, enzyme all can degrade, inactivate, the therefore stability of miRNA；Quickly, high-throughout miRNA chip detection technology；MiRBase data base's is day by day perfect, carries out detection for it as myocardial toxicity biomarker and provides possibility.
Summary of the invention
The purpose of the present invention aims to provide a kind of miRNA purposes in preparing drug-induced heart disease biomarker.
As used in the present invention, the miR-15a-3p of rat is miRNA (miRNA, microRNA) sequence one section ripe, and its base sequence is: 5'-AGGCCAUAUUGUGCUGCCUCA-3'(SEQIDNo.1).
The miR-483-3p of rat is miRNA (miRNA, microRNA) sequence one section ripe, and its base sequence is: 5'-CACUCCUCCCCUCCCGUCUUGU-3'(SEQIDNo.2).
The miR-127-5p of rat is miRNA (miRNA, microRNA) sequence one section ripe, and its base sequence is: 5'-CUGAAGCUCAGAGGGCUCUGAUU-3'(SEQIDNo.3).
The miR-615 of rat is miRNA (miRNA, microRNA) sequence one section ripe, and its base sequence is: 5'-GGGGGUCCCCGGUGCUCGGAUC-3'(SEQIDNo.4).
Specifically, a first aspect of the present invention there is provided miR-15a-3p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-15a-3p is such as shown in SEQIDNo.1, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
A second aspect of the present invention there is provided miR-615 application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-615 is such as shown in SEQIDNo.2, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
A third aspect of the present invention there is provided miR-127-5p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-127-5p is such as shown in SEQIDNo.3, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
A fourth aspect of the present invention there is provided miR-483-3p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-483-3p is such as shown in SEQIDNo.4, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
In a preference, the consumption of described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract will not cause the concentration change of blood plasma Cardiac Troponin-I.
A fifth aspect of the present invention there is provided miR-15a-3p and detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-15a-3p is such as shown in SEQIDNo.1.
A sixth aspect of the present invention there is provided miR-615 and detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-615 is such as shown in SEQIDNo.2.
A seventh aspect of the present invention there is provided miR-127-5p and detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-127-5p is such as shown in SEQIDNo.3.
A eighth aspect of the present invention there is provided miR-483-3p and detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-483-3p is such as shown in SEQIDNo.4.
In a preference, the consumption of described triptolide will not cause the concentration change of blood plasma Cardiac Troponin-I.
The details of various aspects of the present invention will be able to detailed description in chapters and sections subsequently.By hereafter and the description of claim, the feature of the present invention, purpose and advantage will become apparent from.
Accompanying drawing explanation
Fig. 1 uses triptolide 0.1 mg kg of body weight gavage 14 days, the metamorphosis (HE dyeing) of rat heart muscle tissue
Fig. 2 for Cardiac Troponin-I kit measurement various dose triptolide to rat oral gavage 14 days, the concentration of Cardiac Troponin-I
Fig. 3 is for using triptolide 0.1 mg kg of body weight gavage, and real-time quantitative RT-PCR measures the expression trend change of miRNA-615 in blood plasma, miRNA-127-5p, miR-15a-3p and miR-483-3p
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention rather than restriction the scope of the present invention.The reagent used in the following example and raw material all can pass through commercial sources and buy acquisition.The experimental technique of unreceipted actual conditions in the following example, generally conventionally condition or according to manufacturer it is proposed that condition.
Unless otherwise defined, the same meaning that all specialties used in literary composition are familiar with one skilled in the art with scientific words.Additionally, any method similar or impartial to described content and material all can be applicable in the inventive method.The use that preferably implementation described in literary composition and material only present a demonstration.
The features described above that the present invention mentions, or the feature that embodiment is mentioned can be in any combination.The disclosed all features of patent specification can with any composition forms use, each feature disclosed in description, it is possible to any alternative characteristics providing identical, impartial or similar purpose replaces.Therefore except having special instruction, disclosed feature to be only impartial or similar features general example.
Embodiment 1 studies the triptolide toxic action to rat heart
1.1 purposes: observe the Cardiotoxicity of low dosage triptolide.
1.2 experiment materials:
1.2.1 reagent: CMC-Na purchased from Chemical Reagent Co., Ltd., Sinopharm Group, triptolide purchased from good medicine inspection equipment company limited of Shanghai section.
1.2.2 laboratory animal and feedstuff: SD rat, male, body weight (120g～140g), of the right age, healthy.Purchased from Shanghai western pul-Bi Kai laboratory animal company limited, animal credit number: SYXK (Shanghai) 2013-0058.Animal feed irradiation Mus material derives from Shi Lin bio tech ltd, Shanghai, Feed Manufacturing licence: Shanghai raises careful (2008) 04031.
1.2.3 experimental apparatus: superspeed refrigerated centrifuge Centrifuge5810R is purchased from Eppendorf company of Germany
1.3 experimental techniques:
SD rat, 72, male.Body weight is 120g～140g, random packet, divides 6 groups, often group 12,1-6 group dosage respectively 0.0002g/kg, 0.0001g/kg, 0.00005g/kg, 0.000025g/kg, 0.0000125g/kg, CMC-Na group, institute's drug is triptolide (CMC-Na of 0.5% is solvent), dissect after being administered 14 days, take blood and isolating cardiac and other organs.Blood use 3.8% sodium citrate (1:9 uses) anti-condensation, be centrifuged 10 minutes under 4 DEG C and 1000g, take blood plasma, subpackage is stand-by.Heart sections and HE dyeing entrust bio tech ltd of Google, Wuhan to carry out.
1.4 experimental results:
In the HE stained (Fig. 1) of cardiac muscular tissue, it has been found that compared with the control, triptolide causes that rat heart muscle fiber ruptures, and myocardial cell edema is loose, illustrates that the heart of rat is had obvious damage by triptolide.
1.5 brief summaries: low dosage triptolide can cause myocardiac histology pathological changes.
Embodiment 2 studies the concentration sensitivity to myocardial damage of blood plasma Cardiac Troponin-I
2.1 purposes: observe when low dosage triptolide is administered, the sensitivity of classical myocardial damage biomarker Cardiac Troponin-I.
2.2 experiment materials:
2.2.1 reagent: rat heart muscle troponin-i enzyme-linked immunosorbent assay kit purchased from American Lifeassays AB (LifeDiagnostics, Inc.).
2.2.2 laboratory animal and feedstuff: with embodiment 1.2
2.2.3 experimental apparatus: superspeed refrigerated centrifuge Centrifuge5810R is purchased from Eppendorf company of Germany；Microplate reader light absorbs the MolecularDevices company of microplate reader SpectraMax190 purchased from American
2.3 experimental techniques
2.3.1 zoopery:
SD rat, 72, male.Body weight is 120g～140g, random packet, divides 6 groups, often group 12,1-6 group dosage respectively 0.0002g/kg, 0.0001g/kg, 0.00005g/kg, 0.000025g/kg, 0.0000125g/kg, CMC-Na group, institute's drug is triptolide (CMC-Na of 0.5% is solvent), dissect after being administered 14 days, take blood and isolating cardiac and other organs.Blood use 3.8% sodium citrate (1:9 uses) anti-condensation, be centrifuged 10 minutes under 4 DEG C and 1000g, take blood plasma, subpackage is stand-by.
2.3.2 blood plasma Cardiac Troponin-I concentration measures
The operating procedure that the test of blood plasma Myocardial troponin-i is specified according to production company, in 96 orifice plates, is separately added into standard substance or sample diluting liquid 200 μ L, cover film, hatches 2 hours for 25 DEG C.Discard liquid in hole, wash 5 times, blot every hole residual liquid, after adding anti-cTnl antibody diluent 100 μ L, add HRP enzyme enzyme conjugates working solution 100ml, hatch 1 hour for 25 DEG C.Wash 5 times, blot, every hole add zymolyte TMB100 μ L, after overlay film, 25 DEG C hatch 20 minutes after, add stop buffer 100 μ L, terminate reaction.Immediately by the microplate reader OD value in each hole of 450nm wavelength measurement.
2.3.3 data process and statistical analysis
GraphPadPrism5 software or SPSS15.0 software is used to carry out data process, data represent with mean ± standard deviation (mean ± SD), compare employing pairing t-inspection between group and carry out statistical analysis, employing one factor analysis of variance method (one-wayANOVA) analyzing and processing is compared in group, p 0.05 thinks there is significant difference, has statistical significance.
2.4 experimental results:
In the blood plasma of each dosage group rat, compared with matched group, cardiac muscle troponin I there are no significant change (Fig. 2).As can be seen here, only when cardiac muscle major injury, detection blood plasma Cardiac Troponin-I is only significant.As seen from Figure 1, the rat heart that dosage is 0.1mg/kg group has occurred that pathologic changes, but now the concentration of blood plasma Myocardial Troponin I does not change.This illustrates: cardiac muscle troponin I is the biomarker in a kind of heart and injury later stage, is not suitable for the early monitoring index as Radix Tripterygii Wilfordii cardiac toxicity.
2.5 brief summaries: low dosage triptolide will not cause the concentration of blood plasma Cardiac Troponin-I to change.
Embodiment 3 goes out, by miRNA-PCR cDNA microarray, the biomarker that triptolide in blood plasma is cardio-toxicity
3.1 purposes: find new biomarker as the cardio-toxicity early monitoring of Radix Tripterygii Wilfordii property and evaluation index
3.2 experiment materials:
3.2.1 laboratory animal and feedstuff: with embodiment 1.2
3.2.2 experimental apparatus: superspeed refrigerated centrifuge Centrifuge5810R is purchased from Eppendorf company of Germany
3.2.3miRNA-PCR chip (trade name: ExiqonmicroRNAPCRpanelI+II, V3.0) is purchased from Exiqon company of Denmark.
3.3 experimental techniques
3.3.1 zoopery: with 2.2.1 zoopery
3.3.2 blood plasma entrusts upper Haikang to become biological engineering company limited to carry out with the miRNA-PCR chip testing of heart.
3.4 experimental results:
Table 1 shows the rat dosage gavage 14 days with triptolide 50 μ g/kg, uses the test result of miRNA-PCR chip.Wherein, miRNA-615, miRNA-127-5p, miR-15a-3p and miR-483-3p are consistent with the expression in heart at blood, in rising trend or have significance to rise.
The expression of microRNA after the process of table 1 triptolide
3.5 brief summaries: low dosage triptolide can cause that in myocardial cell, multiple miRNA enters blood plasma, and has significance to change.
Embodiment 4 real-time quantitative RT-PCR detects the expression trend change of multiple miRNA in blood plasma.
4.1 purposes: clear and definite clinical diagnosis Best Times also selects corresponding miRNA as diagnostic detection index
4.2 experiment materials:
4.2.1 laboratory animal and feedstuff: with embodiment 1.2
4.2.2miRNA the TaqMan probe miR-615 (AssayID:002353) of probe and reagent: miRNA, miR-127-5p (AssayID:464703_mat), miR-15a-3p (AssayID:474053_mat), miR-483-3p (AssayID:462642_mat) and U6snRNA (AssayID:001973), PCR reagent MIRVANAPARIS40RXNSEACH (article No. AM1556)MicroRNAReverseTranscriptionKit (article No. 4366596), TaqManMastermix:UniversalMasterMixII, noUNG1-Pack (1x5mL) (article No. 4440040), TrizolReagent (15596-026) all purchased from American Lifetechnologies companies.
4.2.3 experimental apparatus: the full-automatic quick beveller of sample is purchased from Shanghai Jing Xin Industrial Co., Ltd.；Ultrasonic cell disruptor is purchased from NingBo XinZhi Biology Science Co., Ltd；Superspeed refrigerated centrifuge Centrifuge5810R is purchased from Eppendorf company of Germany；PCR reverse transcription instrument (thermocycler, Biometra company of Germany)；ABI7500fastPCR amplification instrument.
4.3 experimental techniques:
4.3.1 zoopery: SD rat, 42, male.Body weight is 120g～140g, random packet, divides 7 groups, often group 6, and 1-7 group is dissected after being followed successively by administration respectively 0,1,2,3,5,7,14 day.Institute's drug is triptolide (CMC-Na of 0.5% is solvent), and dosage is 0.1mg/kg.Dissection core dirty, take blood.Blood use 3.8% sodium citrate (1:9 uses) anti-condensation, be centrifuged 10 minutes under 4 DEG C and 1000g, take blood plasma, subpackage, put into liquid nitrogen to be measured.Internal organs first put into IQF in liquid nitrogen, after transfer to-80 DEG C of Refrigerator stores.
4.3.2 real-time quantitative PCR
4.3.2.1 Total RNAs extraction
4.3.2.1.1 the step extracting cardiac muscular tissue's total serum IgE is:
1) weigh cardiac muscular tissue 30-40mg to be put in the EP pipe of 2ml, add 2 steel balls, add the Trizol reagent of 1ml, be put in tissue grinder instrument, beveller adds liquid nitrogen, grinds, take out steel ball.Ultrasound for homogenization, the ultrasonic 4s of interval 2s, super 4-5 time of each sample, room temperature stands 5min.Centrifugal (4 DEG C, 12000rpm, 10min), takes supernatant.
2) often pipe adds 0.2mL chloroform, and fully vibrate 15s, and room temperature stands 15min；
3) being divided into three layers after being centrifuged (4 DEG C, 12000rpm, 15min), transfer about 450 μ L upper strata aqueous phases are to new 1.5mL centrifuge tube, and add equal-volume isopropanol, and for several times, room temperature places 15min in reverse mixing；
4) centrifugal (4 DEG C, 12000rpm, 10min), abandon supernatant, it is seen that white precipitate at the bottom of pipe；
5) 1mL75% washing with alcohol is added once, 4 DEG C, the centrifugal 5min of 12000rpm, abandon supernatant；
6) add 1mL absolute ethanol washing once, after centrifugal, abandon supernatant, super-clean bench inner drying 10min；
7) about 15-25 μ LDEPC treated water dissolution precipitation is added；
8) after 55 DEG C of water-bath 5min, rifle head blows and beats mixing sample repeatedly, therefrom draws 1 μ L and is diluted to 100 μ L, the concentration of ultraviolet determination RNA.
4.3.2.1.2 the experimental procedure of total serum IgE in blood plasma is extracted
Preparation:
Adding 375 μ L2-mercaptoethanols in 2xdenaturing solution, 4 DEG C of preservations, with front 37 DEG C of dissolvings.
21ml dehydrated alcohol, mixing is added in washing liquid 1.
40ml dehydrated alcohol, mixing is added in washing liquid 2/3.
Extract RNA:
1) in blood plasma, isopyknic 2xdenaturing solution is added, mixing, hatch 5min. on ice
2) adding isopyknic acid-phenol:chloroform, vortex 30-60s mixing in above-mentioned mixed liquor, room temperature 12000g is centrifuged 5min, takes supernatant liquid and puts in new EP pipe, remembers volume.
3) eluent be preheating to 95 DEG C stand-by, dehydrated alcohol place room temperature stand-by.
4) in the liquid taken out in 2, add the dehydrated alcohol of its 1.25 times of volumes, mixing.
5) filter element is contained on collecting pipe, mixed liquor is injected in filter element, be not more than 700 μ L every time, centrifugal 30s, abandon filtrate in pipe.
6) filter element is washed by 700 μ L washing liquids 1, centrifugal 15s, abandon washing liquid in pipe.
7) washing filter element 2 times by 500 μ L washing liquids 2/3, centrifugal 15s, abandons washing liquid in pipe every time.Centrifugal collecting pipe 1min is to remove the residual liquid on filter element again, abandons collecting pipe, is placed in by filter element on a new collecting pipe.
8) the elution liquid of 100 μ L95 DEG C is added to filter element, centrifugal 30s, collects RNA.
9) RNA concentration is surveyed.
4.3.2.2 reverse transcription synthesis cDNA
Illustrate according to Reverse Transcription box, the total serum IgE of extraction is carried out reverse transcription.
Each reagent, after thawing on ice, is operated by the reverse transcription system shown in table 2 on ice:
Table 2 reverse transcription system
RNA amount used by each reaction system is 1-10ng.After each reactant liquor and sample are mixed, centrifugal degassing bubble.Putting samples in PCR instrument, reverse transcription reaction program is set to 16 DEG C × 30min, 42 DEG C × 30min, 85 DEG C × 5min, 16 DEG C × 24h.The cDNA that reverse transcription obtains is placed in-80 DEG C of preservations.
4.3.2.3PCR expand
According to description, PCR reaction system is as shown in table 3:
Table 3PCR reaction system
Putting in PCR instrument after sample blending is centrifugal, response procedures is: 95 DEG C × 10min carries out denaturation, and then two-step method (95 DEG C × 15s → 60 DEG C × 60s, 40 circulations) expands.The Ct value of the Ct U6 of genes of interest is standardized by result, calculates gene expression relative quantity according to relative method, adopts formula 2-ΔΔCtCalculate relative fold's change of gene expression.
4.3.3 data process and statistical analysis
GraphPadPrism5 software or SPSS15.0 software is used to carry out data process, data represent with mean ± standard deviation (mean ± SD), compare employing pairing t-inspection between group and carry out statistical analysis, employing one factor analysis of variance method (one-wayANOVA) analyzing and processing is compared in group, p 0.05 thinks there is significant difference, has statistical significance.
4.4 experimental results:
Fig. 3 is the expression trend change detecting blood plasma miRNA-615, miRNA-127-5p, miR-15a-3p and miR-483-3p with real-time quantitative RT-PCR.Blood plasma and heart is taken to solution after rat triptolide (dosage is 0.0001g/kg) 0,1,2,3,5,7,14 day.The miRNA expression real-time quantitative RT-PCR of rat plasma measures.Data represent the mean+SD of at least three independent experiment.
4.5 brief summaries: in rat plasma miRNA-615 and miRNA-127-5p give expression to present early stage, the expression of miR-15a-3p and miR-483-3p then occurs in after above-mentioned 2 kinds of miRNA decline.So, above-mentioned four kinds of miRNA can provide for the diagnosis of Radix Tripterygii Wilfordii property heart disease different phase and select.
Many aspects involved in the present invention have been done as explained above.However, it should be understood that put before not necessarily departing from spirit of the present invention, it can be carried out equivalent change and modification by those skilled in the art, and described change and modification fall into the coverage of the application claims equally.
Claims (10)
1.miR-15a-3p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-15a-3p is such as shown in SEQIDNo.1, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
2.miR-615 application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-615 is such as shown in SEQIDNo.2, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
3.miR-127-5p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-127-5p is such as shown in SEQIDNo.3, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
4.miR-483-3p application in the biomarker of myocardial damage that preparation detection Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract cause, the base sequence of described miR-483-3p is such as shown in SEQIDNo.4, and described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract are with triptolide for active component.
5. the application as described in Claims 1 to 4 any claim, it is characterised in that the consumption of described Radix Tripterygii Wilfordii or Radix Tripterygii Wilfordii extract will not cause the concentration change of blood plasma Cardiac Troponin-I.
6.miR-15a-3p detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-15a-3p is such as shown in SEQIDNo.1.
7.miR-615 detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-615 is such as shown in SEQIDNo.2.
8.miR-127-5p detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-127-5p is such as shown in SEQIDNo.3.
9.miR-483-3p detects the application in the biomarker of the myocardial damage that triptolide causes in preparation, and the base sequence of described miR-483-3p is such as shown in SEQIDNo.4.
10. the application as described in claim 6～9 any claim, it is characterised in that the consumption of described triptolide will not cause the concentration change of blood plasma Cardiac Troponin-I.
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