CN103301459A - Application of anti-heat shock protein 70 (HSP70) in preventing and/or treating hypertensive heart disease and heart failure - Google Patents
Application of anti-heat shock protein 70 (HSP70) in preventing and/or treating hypertensive heart disease and heart failure Download PDFInfo
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Abstract
The invention relates to an application of an anti-heat shock protein 70 (HSP70) in preventing and/or treating hypertensive heart disease and heart failure. An anti-HSP70 neutralizing antibody (Anti-HSP70Ab) of the HSP70 is utilized to inhibit the activity of an extracellular HSP70 molecule on a mouse hypertensive heart disease model, a subacute heart failure model and a chronic hear failure model, so that the animal survival rate can be obviously increased, and cardiac dysfunctions can be partially converted; the HSP70 can be used for inhibiting myocardial hypertrophy caused by hypertension and myocardial fibrosis caused by various reasons, and reducing collagen deposition and alpha-sooth muscle actin (SMA) expression; the HSP70 has functions of regulating the heart local immune microenviroment, inhibiting the inflammatory response and enabling inflammatory reaction to shift towards active inflammation outcome. Results prove that the HSP70 can inhibit the extracellular HSP70 molecular activity and has an obvious treatment function on hypertensive heart disease and heart failure.
Description
Technical field
The present invention relates to suppress the application of activity in preparation treatment hypertensive heart disease and heart failure medicine of the outer heat shock protein 70 of born of the same parents, belong to medical technical field.
Background technology
Congested or chronic heart failure is called for short heart failure, is the eventually end stage of the caused heart disease of a variety of causes, heart contraction and (or) diastolic function obviously reduces so that the heart blood discharge amount can not to satisfy organism metabolism required.Its cause of disease is a lot, mainly comprises coronary heart disease, hypertension, cardiomyopathy, cardiotoxin, valve disease and congenital heart disease etc.According to EPDML statistics, the prevalence of heart failure is made a definite diagnosis 20% death in rear 1 year up to 1-2%, and 5 annual death rate are higher than 50%, and China dies from the hundreds of thousands of them that reaches of heart failure every year.At present, although the Drug therapy of anti-heart failure, interventional therapy and surgical operation etc. have been obtained considerable progress, but these Therapeutic Method can only improve patients with heart failure sings and symptoms, improve its quality of life and delay the progress of the state of an illness, Serious heart Failure is not still had more effective Therapeutic Method except heart transplantation.The core pathological change of hypertensive heart disease comprises myocardial hypertrophy and cardiac fibrosis, also is the main cause that causes chronic heart failure.Research in recent years finds that also inflammatory reaction brought into play important function in hypertensive heart disease and heart failure.Endogenous damage associated molecular pattern (DAMPs) enable mode identification receptor (such as Toll sample receptor), the activated immune inflammatory reaction that cell injury discharges also causes tissue injury, participates in the genesis of heart failure.The molecular mechanism that the reconstruct of further investigation heart failure infection center flesh produces, and find on this basis the new drug target, will have important reference value to the medicine of the multiple heart disease of exploitation treatment.
Heat shock protein 70 (HSP70) is the important member of heat-shock protein family (HSPs), under non-stress state, there is the expression of foundation level in HSP70 in born of the same parents, participate in the processes such as folding, the transhipment of intracellular protein and assembling as molecular chaperones, keeps the stable state of cell.And under the effect of various stressors, the HSP70 that cell discharges then plays a role as a kind of important DAMP molecule, can participate in tissue injury's processes such as oxidative stress, inflammation by activating innate immune system.Present existing research finds that HSPs has participated in the genesis of multiple cardiovascular disease, and for example: in atherosclerotic atherosclerosis plaque forming process, HSP70 promotes macrophage secretion inflammatory factor in the mode of paracrine; In the situation that pressure load increases, the expression of HSP70 significantly increases in laboratory animal aorta and the heart tissue; When the human body hypertension was also found in clinical research, the concentration of HSP70 also increased in the blood circulation, detected the concentration of HSP70 in the blood circulation, damage that can anticipation hyperpietic vitals.In view of the not same-action that HSP70 has with the extracellular in cell, suppress targetedly the activity of the outer HSP70 of born of the same parents, can affect the effect that HSP70 brings into play as a kind of DAMP molecule.
Summary of the invention
High for disease incidences such as hypertensive heart disease and heart failures, mortality rate is high and lack the present situation of clinical active drug Therapeutic Method, the invention provides a kind of new technical scheme, namely suppresses active treatment hypertensive heart disease and the heart failure of the outer HSP70 molecule of born of the same parents.
The inventor of present patent application finds by systematic research, for the core pathological change of hypertensive heart disease and heart failure, utilizes neutrality antibody functionally to block the activity of the outer HSP70 of born of the same parents, can significantly improve the survival rate of animal, improves cardiac function; Suppress the myocardial hypertrophy that hypertensive heart disease causes; Suppress the myocardial fibrosis in hypertensive heart disease and the heart failure process, reduce the expression of collagen deposition and α-SMA; Regulate heart local immunity microenvironment, suppress inflammatory reaction, promote that inflammatory lapses to.
Illustrate that the activity that suppresses the outer HSP70 molecule of born of the same parents has significant therapeutic effect for hypertensive heart disease and heart failure.
" activity that suppresses the outer heat shock protein 70 of born of the same parents " described in the present invention comprises the inhibitory action of utilizing the active of outer heat shock protein 70 molecules of neutrality antibody blocking-up born of the same parents or utilizing other inhibitor, antagonist and other method that the activity of outer this molecule of born of the same parents is produced.
Described hypertensive heart disease and heart failure refer to hypertensive heart disease and the heart failure of people or animal.
With respect to prior art, beneficial effect of the present invention is as follows: the invention provides a kind of new treatment hypertensive heart disease and the scheme of heart failure---suppress the activity of the outer HSP70 of born of the same parents, and evident in efficacy aspect treatment hypertensive heart disease and heart failure, use safety.
Term and abbreviation
The HSP-heat shock protein
Description of drawings
Control: normal mouse group
The Sham:AAC sham operated rats
The Model:Dox/AAC model group
IgG: homotype contrast IgG treatment group
HSP70Ab: anti-HSP70 neutrality antibody treatment group
Fig. 1. anti--HSP70Ab improves the survival rate of heart failure mice
Fig. 2. anti--HSP70Ab significantly improves the cardiac dysfunction of the subacute Heart Failure Model mice that Dox causes
A, the ultrasonic cardiography that the mouse core function is respectively organized in the Dox experiment detects typical figure (M is super)
B, C, the cardiac ejection fraction (EF), left ventricle antetheca thickness (LVAWd) and the interventricular septal thickness (IVSd) that characterize the mouse core function after giving Antybody therapy over time
Fig. 3. anti--HSP70Ab significantly improves the cardiac dysfunction of the Chronic heart failure model mice that Dox causes
A, the ultrasonic cardiography that the mouse core function is respectively organized in the Dox experiment detects typical figure (M is super)
B, C, the ejection fraction, left ventricle antetheca thickness and the interventricular septal thickness that characterize the mouse core function after giving Antybody therapy over time
Fig. 4. anti--HSP70Ab suppresses the myocardial hypertrophy that AAC induces
A, the H﹠amp of the whole section of AAC mouse heart; The E colored graph
B, the HE dyeing of mouse heart tissue is respectively organized in the AAC experiment
C, the M ultrasonic cardiography that the mouse core function is respectively organized in the AAC experiment detects typical figure
D, RT-PCR detects " fetus gene " expression in heart tissue such as ANP, BNP
E, AAC mice interventricular septal thickness cartogram
F, AAC mice left ventricular posterior wall thickness cartogram
G, AAC mouse cardiac myocytes diameter cartogram
H, I, the mRNA of " fetus gene " ANP, BNP analyzes
Fig. 5 .HE demonstration of dyeing, anti--HSP70Ab significantly alleviates the dissolving fracture of the heart tissue cell that Dox causes
Fig. 6. anti--HSP70Ab suppresses the heart tissue fibrosis that attenuates pressure-overload left causes
A, B, anti--HSP70Ab alleviate matter and perivascular fibrosis between the cardiac muscle that AAC induces significantly
D, anti--HSP70Ab reduces heart tissue Collagen volume fraction (p＜0.01) significantly
C, E, immunohistochemical staining and the statistical analysis that mouse heart is organized α-SMA respectively organized in the AAC experiment
F, the statistical analysis of the expression of collagen I mRNA
Fig. 7. anti--HSP70Ab suppresses the heart tissue fibrosis that Dox causes.
A, the Masson dyeing of heart tissue pathological section is respectively organized in the Dox experiment
B, collagen I is respectively organized the expression of heart tissue in the Dox experiment
Fig. 8. anti--HSP70Ab significantly reduces the protein expression that Dox causes heart tissue α-SMA
A, the immunohistochemical staining of α-SMA
B, α-SMA is in the expression of different experiments group heart tissue
Fig. 9. inflammation-associated cytokine is in the expression of Dox different experiments group heart tissue
Figure 10. short inflammation lapses to correlation molecule in the expression of Dox different experiments group heart tissue
The specific embodiment
Below with reference to embodiment invention is described further, but does not limit the scope of the invention.
1. test main agents and laboratory animal
(male, 8-10 is all, 18-23g) available from Beijing Vital River Experimental Animals Technology Co., Ltd. for C57BL/6J mice animal.All animal feedings are in the institute of Materia Medica,Chinese Academy of Medical Sciences Experimental Animal Center, constant temperature and humidity, free diet.
Amycin (Doxorubicin, Dox) is available from Wanle Pharmaceutical Co Ltd, Shenzhen; Anti--IgG antibody is available from Biolegend; Anti--HSP70 antibody is available from Santa Cruz company; Anti--α-SMA antibody is available from Boster company; Employed DAB developer comes Bioisystech Co., Ltd available from Beijing Puli in the Western Blot experiment; Haematoxylin dyeing liquid, Yihong reagent and Masson trichrome stain test kit all step neoplasm scientific and technological development company limited available from Foochow.
2 experimental techniques
2.1 preparation and the packet design of the hypertension induced heart disease model of mice attenuates pressure-overload left (AAC)
The preparation of AAC model
Male C 57 BL/6 J mouse after the animal via abdominal cavity pentobarbital sodium anesthesia (50mg/kg, i.p.), carries out constriction of abdominal aorta or sham-operation on the kidney.The mice dorsal position is fixed in the Mus plate, cut off operation place fur, sterilization after, in the abdominal part median incision, cut skin and flesh layer, otch is 0.5-1cm approximately; After exposing internal organs, stretch into the abdominal cavity with finger, the abdominal cavity is aortal beats, to judge its position, then push to the right side such as Mouse Stomach and mesentery with the sterilization dry cotton ball that is soaked with normal saline and expose ventral aorta before kidney and the spinal column, above renal artery branch, separate ventral aorta 1cm long, be close to the parallel placement of ventral aorta with No. 4 injection needles, with the two together ligation, then extract syringe needle with 1 trumpeter's art stitching thread out, can the constriction ventral aorta.Postoperative is used penicillin 50,000 U/, one week an of lumbar injection every day, in order to avoid infect.
Packet design
Table 1. is anti--and the HSP70 neutrality antibody is to the pharmacodynamic study experiment packet design of AAC model
Mice is divided into 4 groups at random by body weight.Front 1 day of anti--HSP70 antibody group and the capable AAC art of homotype contrast IgG treated animal, postoperative the 2nd, 4,6,13,20,27,34 days dosage tail vein injection HSP70 antibody or IgG according to 200ug/kg.Matched group (Sham) and model group (Model) are injected isopyknic normal saline at same time, and experiment in the 35th day finishes.
2.2 the preparation of subacute heart failure mouse model and packet design
The preparation of subacute heart failure mouse model
Male C57BL/6J (age in 8-10 week, 18-23g) mice, through the sterilization of 75% ethanol abdominal part, single intraperitoneal injection amycin (15mg/kg) modeling, the death condition of every day entry body weight and animal after the modeling finished experiment to the 36th day.
Packet design
Table 2. is anti--the pharmacodynamic study experiment packet design of the subacute heart failure that the HSP70 neutrality antibody causes Dox
The effect of the subacute heart failure that-HSP70 neutrality antibody anti-in order to observe causes Dox, it is 4 groups that experiment is divided into, i.e. normal group (Control), model group (Model), homotype contrast IgG group and anti--HSP70Ab treatment group (HSP70Ab).After the Dox modeling the 10th, 14,18, the tail vein gave injecting normal saline (N.S), IgG or anti--HSP70 neutrality antibody (100 μ g/kg in 25 and 32 days, first dosage doubles), finished experiment on the 36th day, choose 8 time points (the 1st, 3,7,11,13,17,24 and 36 day) in the whole experimentation, treating excess syndrome is tested and is respectively organized the mouse heart tissue and detect.Not modeling of normal group, model group give the normal saline of equivalent and treat.
2.3 the preparation of chronic heart failure mouse model and packet design
The preparation of chronic heart failure mouse model
The C57BL/6 mice is sterilized through 75% ethanol abdominal part, and repeatedly lumbar injection low dosage amycin (3.5mg/kg) is weekly, totally 8 weeks, have a rest a week to recover the mouse bone marrow cells hemopoietic function, and the cardiac function of mice is obviously lacked of proper care in the time of the 64th day, and Chronic heart failure model is successfully prepared.
Packet design
Table 3. is anti--the pharmacodynamic study experiment packet design of the chronic heart failure that the HSP70 neutrality antibody causes Dox
2.4 measure the mice survival rate
Be the same day to calculate in the 0th day from modeling, statistics was respectively organized the laboratory animal death condition and was calculated every day, and certain treated animal dead survival rate do not occur and is calculated as 100%, and certain treated animal all dead survival rates is calculated as 0%.
2.5 ultrasonic cardiography detects the mouse core function
After mice is anaesthetized by 0.32g/kg dosage lumbar injection tribromoethanol, to the processing of losing hair or feathers of its breast abdominal part, dorsal position is fixed in the thermostatic ultrasonic monitor station, use VisualSonics Vevo 770 (Visual Sonics, Canada) ultrasonic system detects: with the 30MHz ultrasonic probe mouse heart long axis direction is carried out the Type B ultrasonic monitoring first, after determining the papillary muscles of left ventricle position, switch to the M pattern and record 10-20 the ultrasonic variation diagram of cardiac cycle; Next adjust again probe orientation (clockwise approximately 90 °) and become the heart short-axis direction and monitor, and 10-20 ultrasonic variation diagram of cardiac cycle of record.Measure respectively the parameters of left ventricular function such as ejection fraction (EF), left ventricle antetheca thickness (LVAWd) and interventricular septal thickness (IVSd) with Vevo770 software.
2.6 pathological section and interpretation of result
Heart is fixed through 4% paraformaldehyde, paraffin embedding, prolongs sagittal direction and cuts into slices.Observe inflammation and fibrosis situation by haematoxylin/Yihong (HE) and Masson dyeing respectively.Use the pathological picture that Spot Advanced 3.0 softwares obtain high-resolution, HE dyeing is observed pathology and is substantially changed.Analyze stained area and the heart tissue area of the rear collagen of Masson dyeing by Image-Pro Plus 5.1.Use Image-Pro Plus 5.1 to measure heart tissue area under the stained area, the visual field of the collagen after each visual field Masson dyeing.The relative amount that represents this collagen with stained area, stained area and visual field heart tissue Area Ratio.10 specimen of every group analysis, each specimen is got 6 visuals field at random, gets average and represents an animal collagen and be organized in relative amount and expression intensity in the heart tissue; Relatively each organizes " collagen absolute area " by the nonparametric variance analysis.
2.7 immunohistochemistry detects the expression of heart tissue cytokine and associated protein
Immunohistochemical staining detects heart tissue α-SMA and expresses variation, and developer DAB (ZLI-9032) is provided by Bioisystech Co., Ltd of China fir Golden Bridge in Beijing.Adopt the SABC method, key step comprises: the conventional dewaxing of tissue slice, aquation; The antigen retrieval of microwave mediation; 3% hydrogen peroxide removal endogenous peroxydase; 0.5%Triton-X100 changes processing thoroughly; The normal serum sealing; Adding 4 ℃ of primary antibodies spends the night; Add biotinylation two anti-, hatch 30min for 37 ℃, ABC complex incubated at room 30min, DAB colour developing; Transparent, mounting.Positive staining is endochylema or karyon brown particle, and negative control replaces with PBS or with the normal IgG of the identical Species origin of primary antibodie.Use the immunohistochemical staining pathological picture that high-resolution color pathological picture and text analytical system Spot Advanced 3.0 obtains high-resolution.8 specimen of every group analysis, each specimen is chosen 15-20 the visual field at random, adopts the positive signal optical density value in each image of Image-Pro Plus 5.1 software analysis, with the relative amount of average optical density value as each sample α-SMA.
2.8 the PCR of plump relevant fetus gene detects
Use " Primer primer 5.0 " software design primer, and carry out homology analysis by Blast software, guarantee its specificity; Get the heart tissue of depositing in the RNA preservation liquid, extract total RNA according to the Trizol method, be dissolved in 20～40ul Erasol water, packing ,-80 ℃ of preservations are stand-by.Get the reverse transcription that 5ug finishes mRNA quantitatively.Utilize RT-PCR, sepharose electrophoresis (EB dyeing), detect plump relevant fetus gene (ANP, BNP etc.) in the heart tissue, with the expression of fibrosis correlation factor (ProCol I etc.).
2.9 Western Blot detects
Extract endochylema and karyon albumen after getting the homogenate of part heart tissue, measure protein concentration by the Branford method.Regulate protein concentration to identical rear adding loading buffer, 96 ℃ of degeneration 6min.Behind the SDS-PAGE electrophoresis, albumen turns the method marking in pvdf membrane by wet; 5% defatted milk powder-TBST solution room temperature sealing 30min adds corresponding primary antibodie 4 degree overnight incubation, and 1%TBST washes film, each 8min, and 3-4 time, add corresponding two anti-incubated at room 1h, wash film 3 times.Specific reaction band is by ECL chemiluminescence system (Amersham Biosciences) colour developing.Calculate each band optical density value with Western blot engram analysis software (Gel pro3.2), analyze multiple proteins and express.
2.10 statistical method
Experimental result mean value ± standard error
Expression, user's difference analysis ANOVA, the suitable method of inspection of the relatively rear variant selection of each group is carried out comparing in twos of many group sample averages.The as a result p of statistical analysis＜0.05 thinks variant, and p＜0.01 is thought significant difference.
3 experimental results
3.1 anti--HSP70Ab improves the animal survival rate
The survival rate result shows, in the chronic heart failure mouse model, except the normal group mice, all the other respectively organize mice in various degree death.During the experiment terminal point, 12 of model group dead mouses, mortality rate are 60%; 5 of HSP70Ab group dead mouses, mortality rate is 25%, has compared significant difference (P＜0.05) with model group; And homotype contrast IgG organizes 11 of dead mouses, and mortality rate is 55%, compares zero difference with model group, shows to utilize anti--HSP70Ab blocking-up HSP70 activity obviously to improve the survival rate (Fig. 1) of heart failure mice.
3.2 anti--HSP70Ab significantly reverses cardiac dysfunction
Clinically, the improvement of Cardiac Function of Patients is the most direct index of reaction Drug therapy effect.Accordingly, we detect the cardiac function of having estimated mice by the mice ultrasonic cardiography.Result from the detection of M type ultrasonic cardiography, subacute and chronic heart failure mouse core chamber obviously enlarges, utilize anti--HSP70Ab neutrality antibody blocking-up HSP70 activity then can partly suppress the expansion of chamber, IgG group mice chamber size (Fig. 2 close to model group, 3), the AAC model mice then owing to hypertension and myocardial hypertrophy, obviously dwindle by model group mouse core chamber, and anti--HSP70Ab treatment can make the ventricular chamber of mice closer to normal mouse.From statistical result, compare with normal mouse, the EF of Dox model group mice, LVAWd, IVSd etc. decrease, and show that Dox causes the left chamber of mice to be shunk and diastolic function obviously descends; And therapeutic resists-and HSP70Ab can partly suppress the deterioration of cardiac function, shows as EF, LVAWd, and the parameters such as IVSd are significantly increased (P＜0.05) (Fig. 2,3) than model group.
3.3 anti--HSP70Ab suppresses the myocardial hypertrophy that hypertensive heart disease causes
35 days histopathology of AAC postoperative and ultrasonic cardiography detection display, the plump degree of model group animal cardiac muscle enlarges markedly (Fig. 4 A) than sham operated rats, be mainly reflected in: interventricular septal thickness (IVS) increases by 50% (Fig. 4 C, 4E) nearly, the left ventricular posterior wall thickness increase is 25% (p＜0.01) (Fig. 4 C, 4F) nearly, and diameter of myocytes has increased nearly 40% (p＜0.01) (Fig. 4 B, 4G).Before mice AAC art and postoperative give the HSP70 antibody capable and suppress significantly interventricular septum, left ventricular posterior wall that hypertension causes and thicken (Fig. 4 C, 4E, 4F) (p＜0.05), diameter of myocytes is compared than model group and is also significantly reduced (p＜0.01); And the degree that gives the mouse cardiac muscle plumpness of homotype contrast IgG has no and alleviates.ANP, BNP etc. " fetus gene " high expressed is one of sign of myocardial hypertrophy, and we have detected the expression of said gene in heart tissue with RT-PCR, result's demonstration, and AAC causes ANP and BNP mrna expression to increase by 2 times; The HSP70 Antybody therapy can reduce the expression (Fig. 4 D, 4H, 4I) (p＜0.05) of above-mentioned fetus gene significantly, and IgG can not reduce the high expressed of said gene.
3.4 anti--HSP70Ab significantly alleviates the dissolving fracture of heart tissue cell
The HE of the cardiac muscular tissue demonstration of dyeing, in the heart failure mice: Normal group myocardial cell form is normal, the kytoplasm clean mark; Model group mouse heart tissue presents kitchen range shape degeneration necrosis, and myocardial cell obvious tumefaction, myocardial cell are degeneration and dissolving fracture in various degree; Anti--HSP70Ab treatment group is than model group have clear improvement (Fig. 5).
3.5 anti--HSP70Ab suppresses the heart tissue fibrosis
Collagen is the most important water-insoluble fibrin in extracellular, is the skeleton that consists of extracellular matrix.Collagen forms the fiber of semi-crystal in extracellular matrix, provide tension stress and elasticity to cell, and works in the migration of cell with in growing.The Etiological of multiple various organ fibrosis is excessive generation and the degraded wretched insufficiency of collagen fiber clinically, thereby cause the extracellular matrix over-deposit, think that at present the fibrosis of cardiac muscular tissue is to make myocardial structural disorderly, cause the infull major reason of diastolic function.Mouse heart is carried out Masson dyeing for we and pathological analysis is found, mice row AAC postoperative 35 days, and the degree that myocardium interstitial fibrosis (Fig. 6 A), heart tissue perivascular fibrosis and tube wall thicken (Fig. 6 B) is significantly higher than the sham operated rats animal.Choose at random 15～20 visuals field the heart tissue Collagen volume fraction is carried out the semi-quantitative analysis discovery, the cardiac fibrosis that the pressure overload causes is 15 times (p＜0.01) under the normal condition, can alleviate significantly matter and perivascular fibrosis (Fig. 6 A, 6B) between the cardiac muscle that AAC induces with the HSP70 Antybody therapy, compare the HSP70 Antybody therapy with the model group animal and can reduce significantly heart tissue Collagen volume fraction (p＜0.01), and homotype contrast IgG treatment group animal hearts tissue fibering degree has no improvement.Also obtained consistent result in the subacute and chronic heart failure mouse model that Dox induces, 36 days hearts form obvious fibrosis after the modeling, and fibrosis area ratio compared with normal group significantly increases; And resist-HSP70Ab treatment group cardiac fibers hamartoplasia obviously alleviates, and the expression of collagen I has also significantly reduced approximately 60%, compares obvious difference (p＜0.05) with model group (Fig. 7).
3.6 anti--HSP70Ab significantly reduces the protein expression of heart tissue α-SMA
α-SMA only expresses in the endochylema than the smooth muscle cell of trunk, bronchial wall in the normal cardiac tissue, and when fibrosis formed, α-SMA can occur in myofibroblast, becomes the sign of fibroblast differentiation.We organize the expression of α-SMA to analyze discovery with methods such as SABC, Western Blot to mouse heart, mice row AAC postoperative 35 days, compare with the sham operated rats mice, the expression of α-SMA has increased nearly 2 times (p＜0.01) (Fig. 6 E) in its heart tissue, with anti--the HSP70 Antybody therapy can reduce the expression (p＜0.05) (Fig. 6 E) of α-SMA significantly, and the IgG treatment group has no change.The experimental result demonstration has also obtained consistent result in the subacute and chronic heart failure mouse model that Dox induces, Dox causes mouse heart to organize the protein expression of α-SMA obviously to increase, and anti--HSP70Ab can significantly reduce this trend (Fig. 8).
3.7 anti--HSP70Ab regulates heart local immunity microenvironment, suppresses inflammatory reaction
Inflammatory reaction has participated in the genesis of heart failure, is considered to the Fibrotic initiating agent of heart tissue.Western Blot analyzes modeling and the short inflammation of rear each the time point heart tissue expression for the treatment of and the expression of anti-inflammatory cytokines, the result shows: after the Dox modeling, some proinflammatory cytokines, expression such as IL-17 obviously increases, reach the highest at the 11st, 13 day its expression, and in anti--HSP70Ab treatment group, the expression of proinflammatory cytokine IL-17 is starkly lower than model group.In contrast, the expression of some anti-inflammatory cytokines such as IL-10 in the expression for the treatment of group apparently higher than the model group mice.By regulating the expression of some cytokines, anti--HSP70Ab has suppressed the inflammatory reaction (Fig. 9) of heart tissue.
3.8 anti--HSP70Ab promotes the inflammation of heart part to lapse to
Lapsing to of inflammation is to guarantee that damaged tissues recovers a significant process of stable state again, and it is not the passive process that is eliminated of inflammation, but an orderly program that is subject to the active of the strict regulation and control of body.Nuclear factor NF κ B-P50,5-lipoxygenase (5-LO), 15-lipoxygenase (15-LO) etc. play a significant role in inflammation lapses to, and have promoted the development of this process.Western Blot has analyzed the expression that each time point these several short inflammation lapse to molecule, discovery is in anti--HSP70Ab treatment group, the expression of the P50 subunit of nuclear factor NF κ B is higher than the expression of P65 subunit, prolongation along with treatment time, the expression for the treatment of group 5-LO, 15-LO, BCL-3 obviously increases, the expression of these molecules has promoted lapsing to of inflammation, impels the organized renewing stable state.According to bibliographical information, cyclo-oxygenase COX-2 has promoted the generation of inflammation then to play a significant role lapsing in the process of inflammation at the initial stage of damage, our result shows the prolongation along with treatment time, the expression of COX-2 presents and reduces first the trend that increases afterwards, and this has also reflected the developing direction of tissue local inflammation from a side.The expression of NO synthase (iNOS) is an important sign of inflammatory reaction severe degree, and our result shows, is resisting-the HSP70Ab treatment group, and the expression of iNOS reduces in time.From these results, anti--HSP70Ab has not only suppressed the inflammatory reaction of heart tissue, also makes inflammatory reaction be offset (Figure 10) towards the direction that lapses to initiatively.
[conclusion]
The active treatment hypertensive heart disease model of the present invention by suppressing the outer HSP70 of born of the same parents and cardiac dysfunction and the myocardial remodelling of subacute Heart Failure Model, Chronic heart failure model animal, find that the activity that suppresses the outer HSP70 of born of the same parents can significantly improve the animal survival rate, part reverses cardiac dysfunction, the myocardial fibrosis that the myocardial hypertrophy that the inhibition hypertensive heart disease causes and a variety of causes cause, regulate heart local immunity microenvironment, suppress inflammatory reaction, promote that inflammatory lapses to.The results show, the activity that suppresses the outer HSP70 of born of the same parents is having broad application prospects aspect preparation treatment hypertensive heart disease and the heart failure medicine.
Claims (2)
1. antibody to heat shock protein 70 prevents and/or treats application in the medicine of hypertensive heart disease in preparation.
2. antibody to heat shock protein 70 prevents and/or treats application in the medicine of heart failure in preparation.
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| CN113546217A (en) * | 2021-07-15 | 2021-10-26 | 中山大学 | Modified acellular myocardial matrix gel and preparation method thereof |
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Non-Patent Citations (3)
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| ASHAKUMARY LAKSHMIKUTTYAMMA: "Interaction between heat shock protein 70 kDa and calcineurin in cardiovascular systems (Review）", 《INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE》 * |
| CAI WF: "Intracellular or extracellular heat shock protein 70 differentially regulates cardiac remodelling in pressure overload mice", 《CARDIOVASC RES》 * |
| 蔡文锋: "调节组织免疫微环境抑制高血压心肌肥厚和心脏纤维化", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
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| CN113546217A (en) * | 2021-07-15 | 2021-10-26 | 中山大学 | Modified acellular myocardial matrix gel and preparation method thereof |
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