CN103911448B - The genetic chip of detection traumatic injury analgesic plaster effect and application thereof - Google Patents
The genetic chip of detection traumatic injury analgesic plaster effect and application thereof Download PDFInfo
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- CN103911448B CN103911448B CN201410132129.8A CN201410132129A CN103911448B CN 103911448 B CN103911448 B CN 103911448B CN 201410132129 A CN201410132129 A CN 201410132129A CN 103911448 B CN103911448 B CN 103911448B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
Abstract
The genetic chip of detection traumatic injury analgesic plaster effect and application thereof, have a probe that RNA, cRNA or the cDNA produced for following gene carries out detecting: 1) inflammatory reaction relevant, 2) wound response is relevant, 3) immune response is relevant, 4) defense response is relevant, 5) angiogenic growth is relevant, 6) energy is correlated with.The genetic chip of the present invention, it is possible to perform well in detecting drug effect and the mechanism of action of traumatic injury analgesic cream.
Description
Technical field
The present invention relates to detect genetic chip and the application thereof of traumatic injury analgesic plaster effect.
Background technology
At present, the medicine effect of China's traumatic injury analgesic cream series products and the research of mechanism of action are nearly at starting
In the stage, document is based on clinical observation on the therapeutic effect, as made the observation of curative effect of blood-circulation and removing blood stasis ointment, injuries from falls as well cream by oneself
The clinical observation etc. for the treatment of acute and closed soft tissue injury.701 traumatic injury analgesic cream also do not carry out deep mechanism
Research.At present, only 1, the document of report, i.e. about the Study of Marketing Tactics of 701 traumatic injury analgesic cream, not
Relate to effect and Mechanism Study.
Summary of the invention
Present invention provide for detecting genetic chip and the application thereof of traumatic injury analgesic plaster effect, it is possible to well
Disclose mechanism of action and the principle of traumatic injury analgesic cream.
For solving the problems referred to above, the technical solution adopted in the present invention is as follows:
The genetic chip of detection traumatic injury analgesic plaster effect, it is characterised in that:
Described genetic chip has the spy that RNA, cRNA or the cDNA produced for following gene carries out detecting
Pin:
1) inflammatory reaction is correlated with: CC CCL11, II type trans-activator, interleukin-11 β, blood
Platelet selects element, p75TNFR；
2) wound response is correlated with: CC CCL11, II type trans-activator, hypoxia inducible factor
1 α, interleukin-11 β, palatelet-selectin, p75TNFR；
3) immune response is correlated with: CC CCL11, II type trans-activator, interleukin-11 β, phosphorus
Fat is creeped enzyme 1, p75TNFR；
4) defense response is correlated with: CC CCL11, II type trans-activator, hypoxia inducible factor
1 α, interleukin-11 β, palatelet-selectin, p75TNFR；
5) angiogenic growth is correlated with: Kruppel like factor 5, hypoxia-inducible factor-1 alpha, interleukin-11 β, blood platelet
Select element；
6) energy is correlated with: Sarcoplasmic reticulum calcium ion ATP enzyme 2, II type trans-activator, hexokinase 3,
Cardiac muscle β-myosin heavy chain 7B, muscle phospho fructokinase, blood platelet class phosphofructokinase, runt are correlated with
Transcription factor 1, Sulfotransferase 1A1.
A kind of method utilizing genechip detection traumatic injury analgesic plaster to imitate, in turn includes the following steps:
A: take a rabbit, then uses the weight method of falling to prepare soft tissue injury model: selection weight is 1kg
Weight, freely fall at vertical direction 20cm of rabbit back leg root soft tissue, it is thus achieved that soft tissue injury
Model；Step B: behind 24 hours that step A completes, measures the scoring of rabbit soft tissue injury, wherein measures
The instrument that valve uses bitterly is tenderness instrument, and red and swollen scope is not less than 1 square centimeter；Step C: in the damage of rabbit
Traumatic injury analgesic cream is sticked at position；Step D: extract RNA at the damage location of rabbit, standby；Step E:
Select DNA chip, with step D extract RNA carry out in situ hybridization, wherein said DNA chip have as
Under RNA, cRNA or cDNA of producing for following gene carry out the probe that detects: 1) inflammatory reaction is correlated with:
CC CCL11 (chemokine (C-C motif) ligand 11), II type trans-activator (class
II transactivator), interleukin-11 β (interleukin-1 beta), palatelet-selectin (selectin, platelet),
P75TNFR (tumor necrosis factor receptor superfamily,
member 1B)；2) wound response is correlated with: CC CCL11 (chemokine (C-C motif) ligand
11), II type trans-activator (class II transactivator), hypoxia-inducible factor-1 alpha
(hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix transcription
Factor)), interleukin-11 β (interleukin-1 beta), palatelet-selectin (selectin, platelet), tumour
Mecrosis factor receptors superfamily member 1B (tumor necrosis factor receptor superfamily, member
1B)；3) immune response is correlated with: CC CCL11 (chemokine (C-C motif) ligand 11),
II type trans-activator (class II transactivator), interleukin-11 β (interleukin-1 beta), phosphatide
Creep enzyme 1 (phospholipid scramblase 1), p75TNFR (tumor
necrosis factor receptor superfamily,member 1B)；4) defense response is correlated with: CC chemotactic factor (CF)
Ligand 11 (chemokine (C-C motif) ligand 11), II type trans-activator (class II
Transactivator), hypoxia-inducible factor-1 alpha (hypoxia-inducible factor 1, alpha subunit (basic
Helix-loop-helix transcription factor)), interleukin-11 β (interleukin-1 beta), blood platelet select
Element (selectin, platelet), p75TNFR (tumor necrosis factor
receptor superfamily,member 1B)；5) angiogenic growth is correlated with: Kruppel like factor 5 (Kruppel-like
Factor 5), hypoxia-inducible factor-1 alpha (hypoxia-inducible factor 1, alpha subunit (basic
Helix-loop-helix transcription factor)), interleukin-11 β (interleukin-1 beta), blood platelet select
Element (selectin, platelet)；6) energy is correlated with: Sarcoplasmic reticulum calcium ion ATP enzyme 2 (sacroplasmic
reticulum Ca2+-ATPase 2), II type trans-activator (class II transactivator), hexokinase 3
(hexokinase 3), cardiac muscle β-myosin heavy chain 7B (myosin, heavy chain 7B, cardiac muscle,
Beta), muscle phospho fructokinase (phosphofructokinase, muscle), blood platelet class phosphofructose swash
Enzyme (phosphofructokinase, platelet), runt associated transcription factor 1 (runt-related transcription
Factor 1), Sulfotransferase 1A1 (sulfotransferase family, cytosolic, 1A, phenol-preferring,
member 1)；Step F: use gene chip scanning instrument to be scanned, obtain scanning signal and data,
After signal and data separate Quantile algorithm are normalized.
In the present invention, preferred scheme is that described F step is: utilize Agilent Microarray Scanner
(Cat#G2565CA, Agilent technologies, Santa Clara, CA, US) is scanned；Wherein scanner
Software design patterns is: Dye channel:Green, Scan resolution=5 μm, PMT 100%, 10%, 16bit；
Then with Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US)
Software reads data and signal；Finally use Gene Spring Software 11.0 (Agilent technologies,
Santa Clara, CA, US) software is normalized, and algorithm used is Quantile.
In the present invention, what Ji metric system pharmaceutical factory of the preferred Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City of traumatic injury analgesic cream is (wide
What Ji metric system pharmaceutical factory, White Cloud Mountain, state Pharmacy stock Co., Ltd White Cloud Mountain) " the 701 traumatic injury analgesic cream " that produces.
Compared with traditional traumatic injury analgesic cream Composition analyzed, the invention have the advantage that the present invention utilizes gene core
Chip technology, uses new zealand rabbit as the acquisition object of soft tissue injury model, then utilizes instrument to carry out
Scanning, detection, data process, and have understood mechanism of action and effect of traumatic injury analgesic cream according to data, with biography
The employing naked eyes of system or microscope are observed, and have the most progressive；This detection method is quick, easy simultaneously,
The experiment parameter filtered out through applicant's great many of experiments and probe combinations, it is possible to testing result is more as the criterion
Really.
Below in conjunction with the accompanying drawings and the present invention is described in further detail by detailed description of the invention.
Accompanying drawing
DNA Microarray Scanning figure used by Fig. 1: embodiment 1；
Box traction substation measured by Fig. 2: embodiment 1；
Scatter diagram measured by Fig. 3: embodiment 1；
Volcano figure measured by Fig. 4: embodiment 1；
Detailed description of the invention
Embodiment 1
A kind of method utilizing genechip detection traumatic injury analgesic plaster to imitate, in turn includes the following steps:
A: take a rabbit, then uses the weight method of falling to prepare soft tissue injury model: selection weight is 1kg
Weight, freely fall at vertical direction 20cm of rabbit back leg root soft tissue, it is thus achieved that soft tissue injury
Model；Step B: behind 24 hours that step A completes, measures the scoring of rabbit soft tissue injury, wherein measures
The instrument that valve uses bitterly is tenderness instrument, and red and swollen scope is not less than 1 square centimeter；Step C: in the damage of rabbit
Traumatic injury analgesic cream is sticked at position, and wherein traumatic injury analgesic cream is how Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City helps public affairs
Pharmaceutical factory (i.e. Baiyunshan Hejigong Medicine Factory of Guangzhou Baiyunshan Pharmaceutical Co., Ltd.) produce " 701 fall
Beat analgesia ointment "；Step D: extract RNA at the damage location of rabbit, standby；Step E: select DNA
Chip, the RNA extracted with step D carries out in situ hybridization, and wherein said DNA chip has following for such as
RNA, cRNA or cDNA that lower gene produces carries out the probe that detects: 1) inflammatory reaction is correlated with: chemokine
(C-C motif)ligand 11、class II transactivator、interleukin-1 beta、selectin,platelet、
tumor necrosis factor receptor superfamily,member 1B；2) wound response is correlated with:
chemokine(C-C motif)ligand 11、class II transactivator、hypoxia-inducible factor 1,
alpha subunit(basic helix-loop-helix transcription factor)interleukin-1 beta、selectin,
platelet、tumor necrosis factor receptor superfamily,member 1B；3) immune response is correlated with:
chemokine(C-C motif)ligand 11、class II transactivator、interleukin-1 beta、
phospholipid scramblase 1、tumor necrosis factor receptor superfamily,member 1B；
4) defense response be correlated with: chemokine (C-C motif) ligand 11, class II transactivator,
hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix transcription
factor)、interleukin-1 beta、selectin,platelet、tumor necrosis factor receptor
superfamily,member 1B；5) angiogenic growth is correlated with: Kruppel-like factor 5, hypoxia-inducible
factor 1,alpha subunit(basic helix-loop-helix transcription factor)、interleukin-1
beta、selectin,platelet；6) energy is correlated with: sacroplasmic reticulum Ca2+-ATPase 2、class
II transactivator、hexokinase 3、myosin,heavy chain 7B,cardiac muscle,beta、
phosphofructokinase,muscle、phosphofructokinase,platelet、runt-related transcription
factor 1、sulfotransferase family,cytosolic,1A,phenol-preferring,member 1；Step F:
Utilize Agilent Microarray Scanner (Cat#G2565CA, Agilent technologies, Santa Clara,
CA, US) it is scanned；Wherein scanner software is set to: Dye channel:Green, Scan
Resolution=5 μm, PMT 100%, 10%, 16bit；Then with Feature Extraction software 10.7
(Agilent technologies, Santa Clara, CA, US) software reads data and signal；Finally use Gene
Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US) software is normalized
Processing, algorithm used is Quantile.
Embodiment 2
A kind of method utilizing genechip detection traumatic injury analgesic plaster to imitate, in turn includes the following steps:
A: take a rabbit, then uses the weight method of falling to prepare soft tissue injury model: selection weight is 1kg
Weight, freely fall at vertical direction 20cm of rabbit back leg root soft tissue, it is thus achieved that soft tissue injury
Model；Step B: behind 24 hours that step A completes, measures the scoring of rabbit soft tissue injury, wherein measures
The instrument that valve uses bitterly is tenderness instrument, and red and swollen scope is not less than 1 square centimeter；Step C: in the damage of rabbit
Traumatic injury analgesic cream is sticked at position, and wherein traumatic injury analgesic cream is how Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City helps public affairs
Pharmaceutical factory (i.e. Baiyunshan Hejigong Medicine Factory of Guangzhou Baiyunshan Pharmaceutical Co., Ltd.) produce " 701 fall
Beat analgesia ointment "；Step D: extract RNA at the damage location of rabbit, standby；Step E: carry with step D
The RNA taken carries out in situ hybridization, wherein said DNA chip have produce for following gene as follows RNA,
CRNA or cDNA carries out the probe detected: 1) inflammatory reaction is correlated with: chemokine (C-C motif) ligand
11、class II transactivator、interleukin-1 beta、selectin,platelet、tumor necrosis factor
receptor superfamily,member 1B；2) wound response is correlated with: chemokine (C-C motif) ligand
11、class II transactivator、hypoxia-inducible factor 1,alpha subunit(basic
helix-loop-helix transcription factor)、interleukin-1 beta、selectin,platelet、tumor
necrosis factor receptor superfamily,member 1B；3) immune response is correlated with: interleukin-1
beta、phospholipid scramblase 1、tumor necrosis factor receptor superfamily,member
1B、Csf2、Tnfrsf6；4) defense response is correlated with: chemokine (C-C motif) ligand 11, class II
transactivator、hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix
transcription factor)、interleukin-1 beta、selectin,platelet、tumor necrosis factor
receptor superfamily,member 1B；5) angiogenic growth is correlated with: Kruppel-like factor 5,
hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix transcription
factor)、interleukin-1 beta、selectin,platelet；6) energy is correlated with: sacroplasmic reticulum
Ca2+-ATPase 2、class II transactivator、hexokinase 3、myosin,heavy chain 7B,cardiac
muscle,beta、phosphofructokinase,muscle、phosphofructokinase,platelet、
runt-related transcription factor 1、sulfotransferase family,cytosolic,1A,
phenol-preferring,member 1；Step F: utilize Agilent Microarray Scanner
(Cat#G2565CA, Agilent technologies, Santa Clara, CA, US) is scanned；Wherein scanner
Software design patterns is: Dye channel:Green, Scan resolution=5 μm, PMT 90%, 10%, 15bit；So
Soft with Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) afterwards
Part reads data and signal；Finally use Gene Spring Software 11.0 (Agilent technologies,
Santa Clara, CA, US) software is normalized, and algorithm used is Quantile.
Embodiment 3
A kind of method utilizing genechip detection traumatic injury analgesic plaster to imitate, in turn includes the following steps:
A: take a rabbit, then uses the weight method of falling to prepare soft tissue injury model: selection weight is 1kg
Weight, freely fall at vertical direction 20cm of rabbit back leg root soft tissue, it is thus achieved that soft tissue injury
Model；Step B: behind 24 hours that step A completes, measures the scoring of rabbit soft tissue injury, wherein measures
The instrument that valve uses bitterly is tenderness instrument, and red and swollen scope is not less than 1 square centimeter；Step C: in the damage of rabbit
Traumatic injury analgesic cream is sticked at position, and wherein traumatic injury analgesic cream is how Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City helps public affairs
Pharmaceutical factory (i.e. Baiyunshan Hejigong Medicine Factory of Guangzhou Baiyunshan Pharmaceutical Co., Ltd.) produce " 701 fall
Beat analgesia ointment "；Step D: extract RNA at the damage location of rabbit, standby；Step E:1) inflammation is anti-
Should be correlated with: chemokine (C-C motif) ligand 11, class II transactivator, interleukin-1 beta,
selectin,platelet、tumor necrosis factor receptor superfamily,member 1B；2) wound
Response is correlated with: Vegefa, Gnrh2, hypoxia-inducible factor 1, alpha subunit (basic
helix-loop-helix transcription factor)、interleukin-1 beta、selectin,platelet、tumor
necrosis factor receptor superfamily,member 1B；3) immune response is correlated with: chemokine (C-C
motif)ligand 11、RT1-Bb、interleukin-1 beta、phospholipid scramblase 1、tumor
necrosis factor receptor superfamily,member 1B；4) defense response is correlated with: chemokine (C-C
motif)ligand 11、class II transactivator、hypoxia-inducible factor 1,alpha subunit
(basic helix-loop-helix transcription factor)、interleukin-1 beta、selectin,platelet、
tumor necrosis factor receptor superfamily,member 1B；5) angiogenic growth is correlated with:
Kruppel-like factor 5、hypoxia-inducible factor 1,alpha subunit(basic
helix-loop-helix transcription factor)、interleukin-1 beta、selectin,platelet；6) energy
Amount is relevant: sacroplasmic reticulum Ca2+-ATPase 2、class II transactivator、hexokinase
3、myosin,heavy chain 7B,cardiac muscle,beta、phosphofructokinase,muscle、
phosphofructokinase,platelet、runt-related transcription factor 1、sulfotransferase
family,cytosolic,1A,phenol-preferring,member 1；Step F: utilize Agilent Microarray
Scanner (Cat#G2565CA, Agilent technologies, Santa Clara, CA, US) is scanned；Its
Middle scanner software is set to: Dye channel:Green, Scan resolution=5 μm, PMT 95%, 10%,
18bit；Then with Feature Extraction software 10.7 (Agilent technologies, Santa Clara,
CA, US) software reads data and signal；Finally use Gene Spring Software 11.0 (Agilent
Technologies, Santa Clara, CA, US) software is normalized, and algorithm used is Quantile.
Embodiment 4
A kind of method utilizing genechip detection traumatic injury analgesic plaster to imitate, in turn includes the following steps: A: take
One rabbit, then uses the weight method of falling to prepare soft tissue injury model: selecting weight is the weight of 1kg, from
Freely fall at vertical direction 30cm of rabbit back leg root soft tissue, it is thus achieved that soft tissue injury model；Step B:
Behind 24 hours that step A completes, measure the scoring of rabbit soft tissue injury, wherein measure the instrument that pain valve uses
For tenderness instrument, red and swollen scope is not less than 1 square centimeter；Step C: the damage location rabbit sticks Die Da town
Cream bitterly, wherein traumatic injury analgesic cream is what Ji (i.e. Guangzhou, metric system pharmaceutical factory of Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City
What Ji metric system pharmaceutical factory, White Cloud Mountain Pharmacy stock Co., Ltd White Cloud Mountain) " the 701 traumatic injury analgesic cream " that produces；Step
Rapid D: extract RNA at the damage location of rabbit, standby；Step E: the RNA extracted with step D is carried out
In situ hybridization, wherein said DNA chip have RNA, the cRNA produced for following gene as follows or
CDNA carries out the probe detected: 1) inflammatory reaction is correlated with: chemokine (C-C motif) ligand 11, class
II transactivator、interleukin-1 beta、selectin,platelet、tumor necrosis factor receptor
superfamily,member 1B；2) wound response is correlated with: Lcn2, Bag3, class II transactivator,
hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix transcription
factor)、interleukin-1 beta、selectin,platelet、tumor necrosis factor receptor
superfamily,member 1B；3) immune response be correlated with: chemokine (C-C motif) ligand 11, Spn,
F3、Ccl5、interleukin-1 beta、phospholipid scramblase 1、tumor necrosis factor
receptor superfamily,member 1B；4) defense response is correlated with: chemokine (C-C motif) ligand
11、class II transactivator、hypoxia-inducible factor 1,alpha subunit(basic
helix-loop-helix transcription factor)、interleukin-1 beta、selectin,platelet、tumor
necrosis factor receptor superfamily,member 1B；5) angiogenic growth is correlated with: Kruppel-like
factor 5、hypoxia-inducible factor 1,alpha subunit(basic helix-loop-helix
transcription factor)、interleukin-1 beta、selectin,platelet；6) energy is correlated with: sacroplasmic
reticulum Ca2+-ATPase 2、class II transactivator、myosin,heavy chain 7B,cardiac
muscle,beta、phosphofructokinase,muscle、phosphofructokinase,platelet、
runt-related transcription factor 1、sulfotransferase family,cytosolic,1A,
phenol-preferring,member 1；Step F: utilize Agilent Microarray Scanner
(Cat#G2565CA, Agilent technologies, Santa Clara, CA, US) is scanned；Wherein scanner
Software design patterns is: Dye channel:Green, Scan resolution=5 μm, PMT 100%, 8%, 14bit；So
Soft with Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) afterwards
Part reads data and signal；Finally use Gene Spring Software 11.0 (Agilent technologies,
Santa Clara, CA, US) software is normalized, and algorithm used is Quantile.
Embodiment 5
According to the detection method of embodiment 1, detect that the target gene of 701 traumatic injury analgesic cream is 302, rise
Gene has 193, and the gene of downward has 109.Target gene carries out GO analyze and show, with inflammation and stress
Relevant down regulation of gene expression, the gene expression of regeneration and multicellular tissue growth is raised.Additionally, capilary
Form gene and ossified gene etc. to raise.The Pathway analysis result of target gene is shown, the letter of differential gene
Number path is mainly relevant, in addition with the inflammatory factor path such as IL-1R signal path, leukocyte adhesion signal path
Also have φt cell receptor signal path, MAPK signal path, nucleic acid acceptor toxicant metabolism path etc. relevant, this
Path is relevant with the action pathway of medicine.
1) chip differential gene data (701 compared with model group)
2) chip differential gene data (model group is compared with negative control group)
The ideograph (Box-whisker Plot) of the box traction substation of Fig. 2 GeneSpring Software on Drawing, is to utilize number
Five statistics according to: minimum of a value, first quartile (25%), median (50%), the 3rd
Quartile (75%) and maximum describe a kind of method of data, and it can also find out that data are roughly
The information such as no have symmetry, point spread of distribution, are especially possible for the comparison to several samples.
Fig. 3 illustrates: the initial data of single Fluorescence chip, through standardization, is converted into log with 2 as the end
Logarithm after, in a two-dimensional direct angle coordinate system plane, draw scatter diagram (scatter plot).Chip data
Scatter diagram be usually used in assessing two groups of data overall distribution central tendencies.In scatter diagram, each point represents chip
On probe points, this position in two dimensional surface is determined by its X-axis coordinate and Y-axis coordinate: X
Axle: this signal value after sample chip Plays.Y-axis: this point is at comparison chip Plays
Later signal value.Fall the point on y=x straight line (neutrality line on figure) in the graphic, and it represents this and visits
Pin mark signal value difference Fold Change=1 in two chips falls in the graphic outside the line of 45 °, bit line both sides
Point, it represents this probe points signal value difference Fold Change > 2 in two chips；A district: 2 times
Up-regulated gene, B district: 2 times of down-regulated genes.
Fig. 4 illustrates: jointly draw volcano with analyzing the P-value value obtained with two factors of Fold change value
Plot volcano figure, for the significant difference of two groups of sample datas of display.In the figure of volcano, one of them is sat
Mark shows the negative log of p-value evolved by T-test, and another one coordinate shows that two conditions compare
Fiducial value after lower log2 conversion.X-axis: Log2 (Fold Change)；Y-axis :-Log10 (p-value)；X
Axle parallel lines: p-value=0.05；Y-axis parallel lines: Fold Change=2；Dark colour part: p-value < 0.05
Fold Change >=2 differential genes.
Rabbit damage model genetic chip result of study shows that model group is 414 with the differential gene of control group,
The gene raised has 346, and the gene of downward has 68.The GO of differential gene analyzes and shows in immunological regulation
The gene (leukocyte recruitment and adhesion, the cytotoxicity of leucocyte mediation) of functional gene and inflammation-related
Increase Deng expression, the gene expression of stress reaction increases, gene of taking action, multicellular tissue growth hormone gene are expressed
Reduce.
Embodiment 6
Detection method to embodiment 1-4 carries out Stability and veracity experiment respectively, and each embodiment carries out five
Secondary experiment, then evaluates its Stability and veracity, and wherein accuracy experimental evaluation data are five experiments
Average, stability experiment utilizes the standard deviation of experimental data to be evaluated, its result such as following table:
| Accuracy (mean value) | Stability (standard deviation) | |
| Embodiment 1 | 90% | 5.2% |
| Embodiment 2 | 83% | 10.1% |
| Embodiment 3 | 85% | 9.5% |
| Embodiment 4 | 86% | 11.3% |
As can be seen from the above table, detection method of the present invention, Stability and veracity is higher, energy
Enough good drug effect and mechanism to traumatic injury analgesic cream is evaluated.
Above-mentioned embodiment is only the preferred embodiment of the present invention, it is impossible to limit present invention protection with this
Scope, the change of any unsubstantiality that those skilled in the art is done on the basis of the present invention and replacement
Belong to scope of the present invention.
Claims (3)
1. the genetic chip of detection traumatic injury analgesic plaster effect, it is characterised in that:
Described genetic chip has the spy that RNA, cRNA or the cDNA produced for following gene carries out detecting
Pin:
1) inflammatory reaction is correlated with: CC CCL11, II type trans-activator, interleukin-11 β, blood
Platelet selects element, p75TNFR；
2) wound response is correlated with: CC CCL11, II type trans-activator, hypoxia inducible factor
1 α, interleukin-11 β, palatelet-selectin, p75TNFR；
3) immune response is correlated with: CC CCL11, II type trans-activator, interleukin-11 β, phosphorus
Fat is creeped enzyme 1, p75TNFR；
4) defense response is correlated with: CC CCL11, II type trans-activator, hypoxia inducible factor
1 α, interleukin-11 β, palatelet-selectin, p75TNFR；
5) angiogenic growth is correlated with: Kruppel like factor 5, hypoxia-inducible factor-1 alpha, interleukin-11 β, blood platelet
Select element；
6) energy is correlated with: Sarcoplasmic reticulum calcium ion ATP enzyme 2, II type trans-activator, hexokinase 3,
Cardiac muscle β-myosin heavy chain 7B, muscle phospho fructokinase, blood platelet class phosphofructokinase, runt are correlated with
Transcription factor 1, Sulfotransferase 1A1.
The genetic chip of detection traumatic injury analgesic plaster the most according to claim 1 effect, it is characterised in that:
Described traumatic injury analgesic cream is the traumatic injury analgesic what Ji metric system pharmaceutical factory of Baiyunshan Pharmaceutics Stock-sharing Co., Ltd., Guangzhou City produces
Cream.
The genetic chip of detection traumatic injury analgesic plaster the most according to claim 1 effect, it falls in detection beats
Application in analgesia ointment drug effect.
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| CN201410132129.8A CN103911448B (en) | 2014-04-02 | 2014-04-02 | The genetic chip of detection traumatic injury analgesic plaster effect and application thereof |
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