CN109846864A - Isoliquiritigenin and its application in the acute lungs liver for the treatment of and brain tissue impairment - Google Patents

Abstract

The present invention provides isoliquiritigenins to treat the application in lungs and the drug of liver damage disease caused by septicopyemia, and, application of the isoliquiritigenin in the drug that treatment wound causes brain tissue damage disease.The drug will be helpful to that disease progression is delayed to deteriorate, and substantially reduce case fatality rate and disability rate, improve life quality.The strong operability for the treatment of, expense is low, can significantly reduce the cost of medical service of patient and medical institutions.

Description

Isoliquiritigenin and its application in the acute lungs liver for the treatment of and brain tissue impairment

Technical field

The present invention relates to a kind of drugs for treating organ injury caused by acute illness, are directed to purulence more particularly to one kind The acute lungs of toxaemia induction and the therapeutic agent of hepar damnification and the acute brain injury as caused by wound.

Background technique

Septicopyemia and traumatic brain injury are two kinds of diseases most commonly seen in Severe acute disease disease.Pyemia (sepsis) refer to systemic inflammatory response syndrome (the systemic inflammatory response caused by infecting Syndrome, SIRS), multiple organ failure is often resulted in, grave danger, pyemic case fatality rate are caused to human life and health Still it is up to 30%~70%.

Traumatic brain injury (traumatic brain injuriy, TBI) refers to the cerebral function as caused by external force or disease Change of science.In recent years, with society development and the vehicles it is prevailing, traumatic brain injury disease incidence increases year by year. In the U.S., 30% traumatic death be by traumatic brain injury caused by, case fatality rate and irreversible neurotrosis occupy high Under not, human health is seriously threatened.

Septicopyemia and traumatic brain injury, which serve not only as independent Severe acute disease, to be occurred, and can be merged simultaneously when more and be deposited The degree of danger of the state of an illness and the difficulty for the treatment of is being further aggravated.Therefore, septicopyemia and traumatic brain damage are understood in depth The pharmaceutical intervention measure new with discovery of the precise mechanism of wound is very necessary.

Current numerous drugs, although as steroid hormone and non-steroid anti-inflammatory drug play certain effect in a short time.But It is to make it in Severe sepsis for the case where crucial pathogenic bacteria of drug-resistant antibiotic ineffective in recent years in terms of the treatment of septicopyemia Effect in disease therapeutic process is greatly limited；The Patients Treated with Steroid of large dosage is to Severe sepsis not energy band Carry out benefit；And low dose steroid therapy effect is unclear, and the curative effect of non-steroidal anti-inflammatory drug also needs further Verifying.Therefore there has been no special anti-inflammatory drugs, and septicopyemia can be effectively relieved at present.

In terms of traumatic brain injury treatment, although multiple organ support therapy technology achieves significant progress, but at present still The progress for having drug that can fast and effeciently delay damage without confirmation.

Septicopyemia is the hot and difficult issue treated at present in Severe acute disease field with organ damage caused by wound at present, is had Effect ground, which is alleviated, even corrects irreversible important organ damage brought by septicopyemia and wound, delays disease progression, saves Life is current urgent problem to be solved.

Summary of the invention

To solve the above-mentioned problems, present inventor has performed sharp studies, as a result, it has been found that: as chalcone, family is important Member, isoliquiritigenin (Isoliquiritigenin, ISO) are widely present in the natural materials such as water fruits and vegetables and herbal medicine.? In terms of pharmacological effect, isoliquiritigenin has the effects that include anti-inflammatory, anti-oxidant and antitumor.Isoliquiritigenin can be by inhibiting inflammation Link is reacted by the lungs and hepar damnification caused by septicopyemia；Alleviated by er stress and autophagocytosis by trauma Caused cerebral injury.

The present invention provides isoliquiritigenin (ISO) as lungs and liver damage disease caused by alleviation and treatment septicopyemia Drug in application；

The present invention provides isoliquiritigenin (ISO) as alleviation and treatment wound and causes answering in the drug of brain tissue damage disease With.

Specifically, the purpose of the present invention is to provide following aspect:

1. application of the isoliquiritigenin in the drug for treating pulmonary lesions disease caused by septicopyemia.

2. application of the isoliquiritigenin in the drug for treating liver damage disease caused by septicopyemia.

3. application of the isoliquiritigenin in the drug that treatment wound causes brain tissue damage disease.

The isoliquiritigenin provided according to the present invention and its application in the acute lungs liver for the treatment of and brain tissue impairment, tool Have it is following the utility model has the advantages that

It is (1) the present invention is intended to provide a kind of for treating the therapeutic agent of organ injury caused by septicopyemia and wound, The advantages such as the drug improves inflammation and/or the damage effect of wound induction is good, has no toxic side effect, and expense is low；

(2) ISO provided by the invention can effectively improve brain injury caused by wound, it will help delays disease progression to deteriorate, shows It writes and reduces case fatality rate and disability rate, improve life quality.

(3) using the strong operability of organ damage caused by ISO provided by the invention treatment septicopyemia and wound, expense It is low, it can significantly reduce the cost of medical service of patient and medical institutions.

Detailed description of the invention

Fig. 1 (A) shows MTT testing result in experimental example 1；

Fig. 1 (B) is shown in experimental example 2 and is induced using ISO alleviation in qRT-PCR method detection primary macrophage by LPS The testing result excessively secreted of inflammatory factor TNFalpha；

Fig. 1 (C) is shown in experimental example 2 and is induced using ISO alleviation in qRT-PCR method detection primary macrophage by LPS The testing result excessively secreted of inflammatory factor IL-6；

Fig. 1 (D), which is shown in experimental example 2, to be alleviated using ISO in Western blot method detection primary macrophage by LPS The testing result of the degradation of the I κ B of induction；

Fig. 1 (E) shows the repetition verification experimental verification that I κ B changes in experimental example 2；

In Fig. 2 (A) A, B, C be shown respectively in experimental example 3 using qRT-PCR technology detection mRNA level in-site ISO alleviate by The testing result that TNF α, IL-6 and MCP-1 excessively secrete in the lung tissue of LPS induction；

In Fig. 2 (B) D, E, F be shown respectively in experimental example 3 using qRT-PCR technology detection mRNA level in-site ISO alleviate by The testing result that TNF α, IL-6 and MCP-1 excessively secrete in the liver organization of LPS induction；

It is shown in Fig. 2 (C) in experimental example 3 and the lungs induced by LPS is alleviated using Hematoxylin-eosin decoration method detection ISO With the testing result of liver organization damage and inflammatory cell infiltration；

It shows in Fig. 2 (D) in experimental example 3 and is lured using ISO alleviation in Western blot method detection lung tissue by LPS The testing result of the degradation of the I κ B led；

It shows in Fig. 2 (E) in experimental example 3 and is lured using ISO alleviation in Western blot method detection liver organization by LPS The testing result of the degradation of the I κ B led；

A is shown in experimental example 4 and is alleviated using Nissl's staining method detection ISO by the brain tissue of wound-induced in Fig. 3 The testing result of the quantity reduction of Nissl corpusculum；B shows slow using Hematoxylin-eosin colouring method detection ISO in experimental example 4 The testing result that solution is damaged by the brain tissue mesencephalic tissue of wound-induced；

A shows mNSS appraisal result in experimental example 5 in Fig. 4；B shows Garcia neurological score result；

A is shown in experimental example 6 using the result of Western blot method detection brain tissue ATF4 in Fig. 5；B shows A's Repeat verification experimental verification；C shows the result of Western blot method detection brain tissue Beclin-1；The repetition that D shows C is tested Card；

A shows the result that immuno-fluorescence assay brain tissue ATF4 is used in experimental example 6 in Fig. 6；B shows brain tissue The result of Beclin-1；

In Fig. 7 (A) C show in experimental example 7 using Western blot method detection SH-SY5Y cell in ISO alleviate by The testing result that the ATF4 of OGD stimulation increases；D shows the repetition verification experimental verification of C；E shows the testing result that CHOP increases；F shows The repetition verification experimental verification of E out；

In Fig. 7 (B) A show in experimental example 7 using Western blot method detection SH-SY5Y cell in ISO alleviate by The testing result that the p62 of OGD stimulation increases；B shows the repetition verification experimental verification of A；C shows the testing result that Beclin-1 increases；D The repetition verification experimental verification of C is shown；

G, which is shown in experimental example 7, in Fig. 7 (C) alleviates the SH- stimulated by OGD using immunofluorescence dyeing method detection ISO The testing result that SY5Y cell ATF4 albumen increases；H shows the testing result that CHOP increases；

E, which is shown in experimental example 7, in Fig. 7 (D) alleviates the SH- stimulated by OGD using immunofluorescence dyeing method detection ISO The testing result that SY5Y cell p62 increases；F shows the testing result that Beclin-1 albumen increases.

Specific embodiment

Present invention will now be described in detail, and the features and advantages of the invention will become more with these explanations It is clear, clear.Any embodiment described herein should not necessarily be construed as preferred or advantageous over other embodiments.Although in the accompanying drawings The various aspects of embodiment are shown, but unless otherwise indicated, it is not necessary to attached drawing drawn to scale.

In the description of the present invention, it should be noted that term " first ", " second ", " third " and " the 4th " are only used for Purpose is described, relative importance is not understood to indicate or imply.

The present invention described below.

Isoliquiritigenin (ISO), molecular weight 256.25, structural formula is as follows:

The study found that isoliquiritigenin can improve the inflammatory signals of lipopolysaccharides (lipopolysaccharideLPS) mediation Access and glycosyloxy deprive the er stress of (Oxygen and glucosedeprivation, OGD) mediation and autophagy is led to Road.

Further study show that ISO can reduce the acute inflammation of the in vitro and in vivo of bacterium inflammation-causing substance LPS induction, it is special It is not that there is potential curative effect in terms of the lungs and hepar damnification that septicopyemia induces；

Wherein, the present invention provides isoliquiritigenins in the drug for treating lipopolysaccharide-induced lungs and liver damage disease Application, especially isoliquiritigenin is in the tissue damage and inflammatory cell infiltration disease for treating lipopolysaccharide-induced lungs and liver Drug in application.

Isoliquiritigenin (ISO) can also be by inhibiting er stress and autophagocytosis to alleviate the damage of the brain as caused by trauma Wound.

Wherein, the present invention provides the brain tissue ATF4 albumen that the isoliquiritigenin is induced in treatment by traumatic brain injury Increase the application in the drug of disease with Beclin-1 albumen.It is lured in addition, additionally providing isoliquiritigenin and being deprived in treatment by glycosyloxy Stress factor ATF4 and CHOP, autophagy factor p62 and the beclin-1 albumen led increase the application in the drug of disease.

The present invention tests two parts by vitro and in vivo and verifies.

In vivo studies level is verified by using models of traumatic brain injury, we carry out pathology to injury tissue Check the damaging pathological change of significant visible brain tissue in discovery brain tissue Hematoxylin-eosin (HE) dyeing and Nissl's staining. And isoliquiritigenin can significantly inhibit trauma induction cerebral injury substantially with the damaging change of local pathology.

In cell level (in vitro test), we verify possible mechanism of action using OGD model.Research finds endoplasmic reticulum Stress important indicator be activating transcription factor 4 (ATF4), CHOP (C/EBP homologous protein), the important indicator of autophagy be p62 and Beclin-1 is significantly increased in modeling group, but is relieved in administration group.

Studies have shown that ISO is by inhibiting er stress and autophagy to improve traumatic brain injury, it is before one kind has exploitation The anti-inflammatory antibody Monoclonal drug of scape.

Experimental example

Experimental example 1

Drug toxicity detection

Incubation of tetrazolium-based colorimetric assay (MTT) the testing inspection hepatic cell line (HepG2) in the ISO of various concentration The variation of lower cell activity: HepG2 cell inoculation is incubated for 24 hours in 96 orifice plates, every 5000 cells at 37 DEG C；It then will be thin After born of the same parents are incubated for 24 hours with isometric 37 DEG C of 1.25,2.5,5,10,20 or 40 μM of ISO or DMSO (dimethyl sulfoxide), carry out Mtt assay.

Cell activity is detected using tetrazolium-based colorimetric assay, analyzes influence of the ISO to liver cell (HepG2), knot Shown in fruit such as Fig. 1 (A), in 1.25 μ Μ, into 40 μ Μ concentration gradients, hepatocyte activity is good, prompts without overt toxicity.

Experimental example 2

Macrophage Model building, inflammatory factor and related pathways detection

The study found that mainly macrophage takes part in inflammatory reaction in lungs or liver organization, therefore in cell level Select macrophage as in vitro study object.

Primary macrophage is extracted:

Primary macrophage extracting method see reference document (J Nutr Biochem 24,146-155, PMID: 22819547, DOI:10.1016/j.jnutbio.2012.03.012).

QRT-PCR technology:

According to Trizol reagent specification, cDNA is synthesized using M-MLV Reverse Transcriptase kit, takes the cDNA reaction solution of preparation 2 μ L are that template (guarantees internal reference and target gene Ct value in 10-40 range, the Ct value is the fluorescence signal in each reaction tube Reach recurring number experienced when the threshold value of setting), reaction system is prepared according to fluorescent quantitative measurement kit specification.PCR Amplification condition are as follows: 95 DEG C, 2min；95 DEG C, 15s are denaturalized, anneal 60 DEG C, 30s, 40 circulations.Each reaction is all provided with 3 multiple holes, Detect the mRNA expression of the Tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in heart tissue.

Primer sequence:

TNF-α:

Upstream 5 '-TGATCCG-CGACGTGGAA-3 ',

Downstream 5 '-ACCGCCTGGAGTTCTGGA-A-3 '；

IL-6:

Upstream 5 '-GAGGATACCACTCCCAACAGACC-3 ',

Downstream 5 '-AAGTGCATCATC-GTTGTTCATACA-3 '；

MCP-1:

Upstream 5 '-TCACCTGCTGCTACTCATTCACCA-3 ',

Downstream 5 '-TACAGCTTCTTTGGGACACCTGCT-3 '；

β-actin (internal reference):

Upstream 5 '-CCGTGAAAAGATGACCCA-GA-3 ',

Downstream 5 '-TACGACCAGAGGCATACAG-3 '.

In primary macrophage, using Real-Time Fluorescent Quantitative PCR Technique (Quantitative RealTime PCR, QRT-PCR) technology confirms, alleviates the excessive secretion by the LPS inflammatory factor TNF-α induced in mRNA level in-site ISO；As a result as schemed Shown in 1 (B).

It in primary macrophage, is confirmed using qRT-PCR technology, alleviates the inflammation induced by LPS in mRNA level in-site ISO The excessive secretion of factor IL-6；As a result as shown in Fig. 1 (C).

Western blot method:

Mouse anesthesia takes lungs or hepatic homogenate after putting to death, 4 DEG C, 12000r/min centrifugation 10min take supernatant, examine horse This brilliant blue detects protein concentration and trim, and Westernblott method detects NF- κ B signal pathway molecule I κ B Degradation Level.Transferring film Primary antibody is incubated overnight afterwards, and TBST buffer is incubated for secondary antibody 1 hour after cleaning, and exposure instrument detects protein expression, measurement purpose and interior Join the integrated absorbance value of band.Wherein, primary antibody is that (dilution ratio 1:1000 is purchased from Cell Signaling biotechnology to I κ B Company)；Secondary antibody is that (it is limited to be purchased from Shanghai assist sage's biotechnology to peroxidase labelling goat anti-rabbit igg by dilution ratio 1:5000 Company)；Internal reference is glyceraldehyde phosphate dehydrogenase (GAPDH).

It is confirmed using Western blot method, as shown in Fig. 1 (D), confirms that ISO is alleviated in protein level and induced by LPS I κ B degradation.The degradation of I κ B make the p65 subunit of NF- κ B from Chromosome migration to nucleus in, with corresponding inflammation phase Correlation gene combines, and starting inflammatory cytokine transcription induces inflammation.Therefore, ISO can alleviate the inflammation induced by LPS.

And the repetition verification experimental verification of the degradation of I κ B has been carried out, as a result as shown in Fig. 1 (E), wherein * P < 0.05, * * P < 0.01, compared with LPS group.

Experimental example 3

Septicopyemia causes lungs and the building of hepar damnification animal model and intervenes

Cleaning grade male C57BL/6 mouse 30,10~12 week old, 23~25g of weight is purchased from the experiment of Beijing dimension tonneau China Zoo technical Co., Ltd uses LPS (20mgkg after adapting to environment 1 week^-1Tail vein injection) induction septicopyemia cause lung Dirty and liver damage model.

Experimental group:

Model group (10), using above-mentioned modeling mode；

Administration group (10) gives ISO (20mgkg in 15 minutes after LPS induction^-1Tail vein injection)；

Control group (10), tail vein gives the physiological saline of same volume.

Tumor necrosis factor-in lungs and liver organization is detected using real-time fluorescence quantitative PCR (qRT-PCR) The mRNA of α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) are expressed, Western blot Method detects I κ B Degradation Level in lungs and liver organization nuclear transcription factor-kappa B (NF- κ B) signal path.

It is tested using the qRT-PCR technology in embodiment 2, wherein

It is confirmed using qRT-PCR technology, alleviates TNF-α, IL-6 in the lung tissue induced by LPS in mRNA level in-site ISO With the excessive secretion of MCP-1.As a result as shown in A, B and C in Fig. 2 (A).

It is confirmed using qRT-PCR technology, alleviates TNF-α, IL-6 in the liver organization induced by LPS in mRNA level in-site ISO With the excessive secretion of MCP-1；As a result as shown in D, E and F in Fig. 2 (B).

Lungs and liver organization pathologic damage are observed using Hematoxylin-eosin decoration method, as a result such as G institute in Fig. 2 (C) Show, ISO is prompted to alleviate by the LPS lung tissue damage induced and inflammatory cell infiltration；

H is prompted in Fig. 2 (C), and ISO is alleviated by the LPS liver organization damage induced and inflammatory cell infiltration.

It is tested using the Western blot method in embodiment 2, in which:

Lung tissue level is confirmed using Western blot method, as shown in Fig. 2 (D), confirms ISO in protein level Alleviate the degradation by the LPS I κ B induced.

Liver organization level is confirmed using Western blot method, as shown in Fig. 2 (E), confirms ISO in protein level Alleviate the degradation by the LPS I κ B induced.

Experimental example 4

The building of traumatic brain injury animal model and intervention

Cleaning grade male C57BL/6 mouse 30,10~12 week old, 23~25g of weight is purchased from the experiment of Beijing dimension tonneau China Zoo technical Co., Ltd establishes mouse traumatic brain injury (TBI) mould using freely falling body punch method after adapting to environment 1 week Type.

Model building method:

Mouse is after chloraldurate (3ml/kg) intraperitoneal injection of anesthesia, calvarium portion preserved skin, complexing iodine disinfectant disinfection, fixed In stereotaxic apparatus, in sagittal midsection scalp, exposed fascia and blunt separation, removing appear left side skull, in bregma About 1.5mm, middle line side 1.5mm afterwards, are carefully bored with dental burr and open 3mm bone window, guarantee that endocranium is complete.Then adjustment cranium brain is beaten Device is hit, diameter 2.5mm 20g striker face bone window center, control strike depth 2mm, the vertical strike from 20cm are made.Modeling at Function standard are as follows: after strike, the transient tic of limbs of mouse, apnea, but can voluntarily alleviate after the several seconds.Postoperation hemostatic, with bone Wax is closed bone window, sutures scalp.Postoperative detection mouse breathing, heartbeat give isothermal holding, keep anus temperature (37.0 ± 0.5) DEG C, Until mouse is awake.

Experimental group:

Model group, using above-mentioned modeling mode；

Administration group gives ISO (20mgkg in 15 minutes after modeling^-1Tail vein injection)；

Control group, tail vein give the physiological saline of same volume.

Cerebral morphology detection:

Preference pattern group, administration group and control group mice each 10 put to death all mouse, stone after anesthesia in 24 hours after administration Wax embeds brain tissue, detects cerebral morphology by Nissl (Nissl) colouring method and Hematoxylin-eosin (HE) colouring method Change, fluorescence microscopy microscopic observation neurotrosis situation.

As shown in A in fig. 3, in model group, the quantity of Nissl corpusculum is reduced Nissl coloration result, prompt nerve cell by To damage；In administration group, ISO improves to be reduced by the quantity of Nissl corpusculum in the brain tissue of wound-induced.

HE coloration result as shown by B in fig. 3, the results show that the loose fracture of model group mesencephalic tissue；And in administration group, ISO improves to be damaged by the brain tissue mesencephalic tissue of wound-induced.

Experimental example 5

The assessment of traumatic brain injury animal model nervous function damage

Model group and administration group mouse each 6 in choice experiment example 4, according to the above method modeling and the 1st after being administered, 3,7, 14,21 and 28 day set time do neurological deficits score (modified neurological severity score, MNSS), food and water are during which supplemented on time, regularly replace padding.

According to mNSS marking scales, is judged using double-blind study and record score value.MNSS standards of grading from mouse movement, sense Feel, balanced capacity and reflection case etc. are assessed, score value 0 (most serious) -18 (normal).

Preference pattern group and administration group mouse each 6, according to the above method modeling and the 1st, 3,7,14,21 and 28 day after being administered Set time does Garcia neurological score, during which supplements food and water on time, regularly replaces padding.

According to Garcia marking scales, is judged using double-blind study and record score value.Garcia standards of grading are before and after mouse Limb harmony, balance etc. are assessed, score value 0 (most serious) -21 (normal).

As a result as shown in A and B in Fig. 4, assessed by two different nervous function damages it is found that ISO can improve by The brain tissue impairment of wound-induced.

Experimental example 6

The detection of Protein Expression of Brain level

Western blot method:

Preference pattern group, administration group and control group each group mouse each 10 put to death all small after anaesthetizing within 24 hours after administration Mouse takes brain tissue homogenate, and 4 DEG C, 12000r/min centrifugation 10min take supernatant, and Coomassie brilliant blue detection protein concentration is simultaneously matched Flat, Westernblot method detects the expression of binding protein ATF4 and endoplasmic reticulum autophagy proteins Beclin-1.

By A in Fig. 5 and C it is found that ISO can alleviate brain tissue ATF4 albumen and the increasing of Beclin-1 albumen by TBI induction It is high.

Histogenic immunity fluorescence detection method: by the binding protein ATF4 of immuno-fluorescence assay brain tissue and interior The expression of matter net autophagy proteins Beclin-1 changes, specific as follows:

(phosphate-buffered salt is molten in PBS for take model group, administration group and control group each group Mice brain tissues tissue freezing section Liquid) in impregnate 15min, with 1%BSA (fetal calf serum) close 30min；Glass slide and first antibody ATF4, Beclin-1 (dilution Ratio is 1:200, is purchased from abcam company) it is placed in wet box after 4 DEG C of overnight incubations, it is placed at room temperature for 2h, PBS washing；It is used in darkroom Two anti-FITC of fluorescence and TRITC (dilution ratio 1:1000 is purchased from abcam company) incubation at room temperature 1h, PBS washing；It reuses DAPI (4', 6- diamidino -2-phenylindone) is incubated for 5min, PBS washing；Plus anti-fluorescence quenching (purchased from green in darkroom Skies company), mounting；Image is obtained using Laser Scanning Confocal Microscope.

As a result as shown in A and B in Fig. 6, it is known that, ISO can alleviate by TBI induction brain tissue ATF4 albumen and Beclin-1 albumen increases.

Er stress is prompted to increase when ATF4 albumen increases, and Beclin-1 albumen increases prompt autophagy and increases.Therefore, ISO can alleviate brain tissue er stress and autophagy.

Experimental example 7

The building of traumatic brain injury cell model and intervention

OGD induction SH-SY5Y endocytoplasmic reticulum stress and autophagy

Western blot method:

1.2×10⁶A SH-SY5Y cell is with DMEM culture medium in 37 DEG C, 5%CO₂Under the conditions of cultivate, more every 24 hours New culture solution is that tested (final concentration of 10 μM) of ISO of 70% or so addition pre-processes 2 hours to cell density, and OGD is continued with 6 hours.Collect group of cells lysate, 4 DEG C, 12000r/min centrifugation 10min take albumen supernatant, Coomassie brilliant blue detects albumen Concentration and trim measure er stress factors A TF4 and CHOP albumen and autophagy factor p62 and beclin-1 albumen water It is flat.

As a result as shown in Fig. 7 (A) and Fig. 7 (B), confirm that ISO alleviates the stress factor ATF4 induced by OGD in protein level With the raising of CHOP, autophagy factor p62 and beclin-1 albumen.

It has also carried out repeating verification experimental verification, as a result as shown in Fig. 7 (A) and Fig. 7 (B), wherein P < 0.01 * *, with OGD group ratio Compared with.

Immunofluorescence dyeing method:

1.2×10⁶A SH-SY5Y cell is with DMEM culture medium in 37 DEG C, 5%CO₂Under the conditions of cultivate, more every 24 hours New culture solution is that tested (final concentration of 10 μM) of ISO of 70% or so addition pre-processes 2 hours to cell density, and OGD is continued with 6 hours.Using immunofluorescence dyeing method, er stress factors A TF4 and CHOP and endoplasmic reticulum autophagy factor p62 is detected With beclin-1 protein level.

As a result as shown in Fig. 7 (C), ISO is alleviated cell ATF4 and the CHOP albumen induced by OGD and is increased；

As a result as shown in Fig. 7 (D), ISO is alleviated cell p62 and the Beclin-1 albumen induced by OGD and is increased.

Traumatic brain injury can be alleviated and treat by demonstrating ISO again in cell level.

It is described the invention in detail above in conjunction with detailed description and exemplary example, but these explanations are simultaneously It is not considered as limiting the invention.It will be appreciated by those skilled in the art that without departing from the spirit and scope of the invention, Can be with various equivalent substitutions, modifications or improvements are made to the technical scheme of the invention and its embodiments, these each fall within the present invention In the range of.Scope of protection of the present invention is subject to the appended claims.

Claims (9)

1. application of the isoliquiritigenin in the drug for treating pulmonary lesions disease caused by septicopyemia.

2. application according to claim 1, which is characterized in that the isoliquiritigenin is treating lipopolysaccharide-induced lungs damage Application in the drug of evil disease.

3. application according to claim 2, which is characterized in that the isoliquiritigenin is treating lipopolysaccharide-induced lungs group Knit the application in the drug of damage and inflammatory cell infiltration disease.

4. application of the isoliquiritigenin in the drug for treating liver damage disease caused by septicopyemia.

5. application according to claim 4, which is characterized in that the isoliquiritigenin is treating lipopolysaccharide-induced liver damage Application in the drug of evil disease.

6. application according to claim 5, which is characterized in that the isoliquiritigenin is treating lipopolysaccharide-induced liver group Knit the application in the drug of damage and inflammatory cell infiltration disease.

7. application of the isoliquiritigenin in the drug that treatment wound causes brain tissue damage disease.

8. application according to claim 7, which is characterized in that the isoliquiritigenin is induced in treatment by traumatic brain injury Brain tissue ATF4 albumen and Beclin-1 albumen increase the application in the drug of disease.

9. application according to claim 8, which is characterized in that the isoliquiritigenin is answered in treatment by glycosyloxy is deprivation induced Swash factors A TF4 and CHOP, autophagy factor p62 and beclin-1 albumen increases the application in the drug of disease.