JP2008001646A - Agent for treatment of cardiac disease - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a medicine reducing the oxidation stress on the heart and suppressing the apoptosis of myocardial cell occurring subsequent to oxidation stress. <P>SOLUTION: The medicine for the prevention and/or treatment of cardiac diseases such as ischemic cardiopathy, heart failure and myocardial disease contains CCN1 protein or a gene encoding CCN1 protein as an active component. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a medicament for the prevention and / or treatment of heart disease.

  Oxidative stress is an important factor in cardiac dysfunction in acute and chronic heart diseases such as ischemia reperfusion injury in acute myocardial infarction and remodeling in chronic heart failure. Therefore, suppressing oxidative stress in the heart and subsequent apoptosis of cardiomyocytes is promising as a means for preventing and treating heart disease.

On the other hand, CCN1 (sometimes called Cyr61: cysteine rich 61) is one of six proteins belonging to the CCN family (Cyr61, CTGF (connective tissue growth factor), NOV (nephroblastoma overexpressed), WISP-1, WISP-2, and It is one of WISP-3), a secreted protein of 43 kD. About this CCN1, there are some reports as a cancer-related gene (nonpatent literature 1 thru | or 3). It is also known that CCN1 binds to an extracellular matrix and exerts an angiogenic action. More specifically, CCN1 promotes the proliferation and migration of vascular endothelial cells (Non-Patent Document 4), CCN1 causes angiogenesis in the rat cornea (Non-Patent Document 5), and CCN1 cDNA is used as a lower limb artery. It has been reported that blood circulation improves when administered to an obstructed rabbit (Non-patent Document 6). Recently, it has been reported that the expression of CCN1 is increased in the heart of ischemic heart disease (Non-patent Document 7). However, there is no report on the direct action of CCN1 on cardiomyocytes and the angiogenesis action in the heart, and no attempt has been made to use CCN1 for the treatment of heart diseases such as ischemic heart disease.
Oncogene, 21, 8178, 2002 J. Biol. Chem., 279, 53087, 2004 Clin. Cancer Res., 11, 7243, 1005 Mol. Cell. Biol., 16, 1326, 1996 Proc. Natl. Acad. Sci. USA, 95, 6355, 1998 Hum. Gene Ther., 13, 1461, 2002 Circulation, 109, 2227, 2004

  An object of the present invention is to provide a medicament for the prevention and / or treatment of heart disease. More specifically, it is an object of the present invention to provide a medicament that can reduce oxidative stress in the heart and suppress cardiomyocyte apoptosis that occurs following oxidative stress.

  The present inventors have found that CCN1 protein is specifically expressed in the embryonic heart in the process of cloning a gene encoding a secretory factor expressed in the developing heart. This finding suggests that CCN1 protein may be involved in the formation of heart and coronary arteries during embryonic period. The present inventors further studied the role of CCN1 protein in the heart, and CCN1 protein protects cardiomyocytes from oxidative stress and suppresses apoptosis, exerts angiogenic action in the heart, and improves cardiac function It was found to have an action. The present invention has been completed based on the above findings.

That is, the present invention provides a medicament for the prevention and / or treatment of heart disease comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient. According to a preferred aspect of the invention, there is provided the above medicament wherein the heart disease is ischemic heart disease, preferably angina pectoris or acute myocardial infarction, and the above medicament wherein the heart disease is heart failure or cardiomyopathy.
Further, according to the present invention, a cardiac function improving agent comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient, a cardiac angiogenesis promoting agent comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient, from oxidative stress A protective agent for the heart, comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient, an inhibitor of cardiac apoptosis caused by oxidative stress, and a gene encoding CCN1 protein or CCN1 protein Inhibitors are provided as active ingredients.

Furthermore, according to the present invention, there is provided a method for the prevention and / or treatment of heart disease, comprising the step of administering CCN1 protein to mammals including humans, preferably comprising the step of locally administering CCN1 protein to the heart. Provided. Also provided by the present invention is a method for preventing and / or treating heart disease, which comprises the step of administering a vector containing a gene encoding CCN1 protein to mammals including humans. According to a preferred aspect of the present invention, there is provided a method comprising the step of locally administering the vector to the heart.
Furthermore, the use of CCN1 protein or a gene encoding CCN1 protein for the manufacture of the above medicament is also provided by the present invention.

  The medicament of the present invention is useful as a medicament for the prevention and / or treatment of heart diseases such as ischemic heart disease. In particular, the medicament of the present invention has an action of protecting myocardial cells from oxidative stress and suppressing apoptosis, and also exerts an angiogenic action in the heart to improve cardiac function. It is possible to effectively treat an ischemic heart disease.

  CCN1 protein which is an active ingredient of the medicament of the present invention is also known as Cyr61 (cystein-rich 61) protein, and the amino acid sequence of human CCN1 protein is described in GenPept NP_001545. In addition, the existence of a gene encoding CCN1 protein (hereinafter sometimes referred to as CCN1 gene) is also known, and the nucleic acid sequence of human CCN1 gene is described in GenBank NM_001554. Accordingly, both protein and gene information is available to those skilled in the art.

  As an active ingredient of the medicament of the present invention, in addition to the wild-type CCN1 protein derived from a mammal, one or several amino acids are substituted, inserted, and / or deleted in the amino acid sequence of the wild-type CCN1 protein. Alternatively, a modified CCN1 protein that can exhibit substantially the same action as the wild-type CCN1 protein in a mammalian organism may be used. When the medicament of the present invention is applied to humans, one or several amino acids are substituted, inserted, and / or deleted in the human-derived wild-type CCN1 protein or the amino acid sequence of human-derived wild-type CCN1 protein. Therefore, it is preferable to use a modified CCN1 protein capable of exhibiting substantially the same action as human-derived wild-type CCN1 in the human body. When referred to herein as “CCN1 protein”, both wild-type CCN1 protein and modified CCN1 protein are included.

  In the present specification, when referring to an amino acid sequence having 1 to several amino acid deletions, substitutions and / or additions, the range of “1 to several” is not particularly limited, but for example, 1 to 20, The number is preferably 1 to 10, more preferably 1 to 7, further preferably 1 to 5, and particularly preferably about 1 to 3. The amino acid sequence of the modified CCN1 protein has, for example, 70% or more homology with the amino acid sequence of the wild-type protein, preferably 80% or more, more preferably 85% or more, more preferably 90%, Particularly preferably, it has a homology of 95% or more.

  As an active ingredient of the medicament of the present invention, in addition to the wild-type CCN1 gene derived from a mammal, 1 to several amino acids are substituted, inserted, and / or deleted in the amino acid sequence of the wild-type CCN1 protein. A DNA encoding a modified CCN1 protein capable of exhibiting substantially the same action as the wild-type CCN1 protein in a mammalian organism may be used. When the medicament of the present invention is applied to humans, one or several amino acids are substituted, inserted, and / or deleted in the amino acid sequence of human-derived wild-type CCN1 gene or human-derived wild-type CCN1 protein. It is preferable to use a DNA encoding a modified CCN1 protein (hereinafter sometimes referred to as a modified CCN1 gene) that can exhibit substantially the same action as a human-derived wild-type CCN1 protein in the human body. . In the present specification, the term “CCN1 gene” includes both a wild-type CCN1 gene and a modified CCN1 gene.

  For example, a DNA that hybridizes under stringent conditions to the wild-type CCN1 gene can be used as the modified CCN1 gene. Such DNA may have a base sequence in which one to several bases are substituted, inserted, and / or deleted in the nucleic acid sequence of the wild-type CCN1 gene. Although the range of “1 to several” is not particularly limited, for example, 1 to 40, preferably 1 to 30, more preferably 1 to 20, more preferably 1 to 10, more preferably 1 to The number is 5, particularly preferably about 1 to 3. Examples of DNA that hybridizes under stringent conditions include DNA having a certain homology with the base sequence of the wild-type CCN1 gene, for example, 70% or more, preferably 80% or more, more preferably 90% or more. More preferred is DNA having a homology of 93% or more, particularly preferably 95% or more, and most preferably 98% or more.

  In the present specification, the “base sequence that hybridizes under stringent conditions” means, for example, colony hybridization method, plaque hybridization method, Southern blot hybridization method, etc. using wild-type CCN1 gene DNA as a probe. For example, using a filter on which colony or plaque-derived DNA or a fragment of the DNA is immobilized at 65 ° C. in the presence of 0.7 to 1.0 M NaCl. After hybridization, DNA etc. that can be identified by washing the filter under a condition of 65 ° C. using 0.1 to 2 × SSC solution (1 × SSC solution is 150 mM sodium chloride, 15 mM sodium citrate) Can be mentioned. Hybridization is a method described in Molecular Cloning: A laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1989 (hereinafter sometimes abbreviated as "Molecular Cloning 2nd Edition"). It can be performed according to.

  The method for obtaining the wild-type CCN1 protein or the above-mentioned modified protein, or the wild-type CCN1 gene or the modified CCN1 gene is not particularly limited. Prepare appropriate probes and primers using the wild-type CCN1 protein amino acid sequence information or wild-type CCN1 gene nucleic acid sequence information described in the above publications, and screen eukaryotic cDNA libraries using them. By doing so, the wild type CCN1 gene can be isolated. For example, the wild type CCN1 gene can be obtained by PCR. PCR can be performed using a pair of primers designed to amplify the wild type CCN1 gene using a chromosomal DNA library or cDNA library derived from a eukaryote as a template. PCR reaction conditions can be set as appropriate. For example, one cycle of a reaction step consisting of 94 ° C for 30 seconds (denaturation), 55 ° C for 30 seconds to 1 minute (annealing), and 72 ° C for 2 minutes (extension) As an example, there may be mentioned conditions for carrying out the reaction at 72 ° C. for 7 minutes after 30 cycles. The amplified DNA fragment can then be cloned into a suitable vector that can be amplified in a host such as E. coli.

  Techniques such as preparation of the probe or primer, construction of a cDNA library, screening of the cDNA library, and cloning of the target gene are well known and commonly used by those skilled in the art. For example, Molecular Cloning 2nd Edition, Current Protocols in Molecular Biology, Supplements 1-38, John Wiley & Sons (1987-1997) etc. can be followed.

  The modified CCN1 gene can be prepared by any method known to those skilled in the art, such as chemical synthesis, genetic engineering techniques, or mutagenesis, based on the amino acid sequence information. For example, the DNA can be obtained using a method of contacting a wild-type CCN1 gene with a drug that is a mutagen, a method of irradiating with ultraviolet rays, a genetic engineering method, or the like. Site-directed mutagenesis, which is one of genetic engineering techniques, is useful because it can introduce a specific mutation at a specific position. For example, Molecular Cloning 2nd Edition, Current Protocols in. It can be performed according to the method described in Molecular Biology and the like.

  The method for obtaining the wild-type CCN1 protein or modified CCN1 protein is not particularly limited, but typically, the wild-type CCN1 gene or modified CCN1 gene is obtained by the above-described method, and this DNA is introduced into an appropriate expression system. It can be obtained by expressing. The wild type CCN1 gene or the modified CCN1 gene can be used by inserting it into an appropriate vector. The type of vector is not particularly limited. For example, a vector that replicates autonomously (eg, a plasmid), a vector that is integrated into the host cell genome when introduced into the host cell, and replicated with the integrated chromosome, etc. Can be used. Preferably, an expression vector can be used, and in the expression vector, it is desirable that the wild-type CCN1 gene or the modified CCN1 gene is functionally linked to elements necessary for transcription (for example, a promoter or the like). A promoter is a DNA sequence that exhibits transcriptional activity in a host cell, and can be appropriately selected depending on the type of host.

Promoters operable in bacterial cells include the Bacillus stearothermophilus maltogenic amylase gene, the Bacillus licheniformis alpha-amylase gene, and the Bacillus amyloliquefati. Enns-BAN amylase gene (Bacillus amyloliquefaciens BAN amylase gene), Bacillus subtilis alkaline protease gene (Bacillus subtilis alkaline protease gene) or promoters of Bacillus pumilus-xylosidase gene (Bacillus pumilus xylosldase gene), or the phage lambda P _R or P _L promoters, lac of E.coli, trp, or the like tac promoter and the like.

  Examples of promoters that can operate in mammalian cells include the SV40 promoter, the MT-1 (metallothionein gene) promoter, or the adenovirus 2 major late promoter. Examples of promoters that can operate in insect cells include polyhedrin promoter, P10 promoter, autographa calicornica polyhedrosis basic protein promoter, baculovirus immediate early gene 1 promoter, or baculovirus 39K delayed early gene There are promoters. Examples of promoters that can operate in yeast host cells include promoters derived from yeast glycolytic genes, alcohol dehydrogenase gene promoters, TPI1 promoters, ADH2-4c promoters, and the like. Examples of promoters that can operate in filamentous fungal cells include the ADH3 promoter or the tpiA promoter.

  In the above expression vector, the wild-type CCN1 gene or the modified CCN1 gene may be operably linked to an appropriate terminator as necessary. The expression vector may have elements such as a transcription enhancer sequence (eg, SV40 enhancer). The expression vector may have a DNA sequence that allows the vector to replicate in the host cell. An example is the SV40 origin of replication when the host cell is a mammalian cell.

  The expression vector may contain a selectable marker. As a selectable marker, for example, a gene whose complement is lacking in the host cell such as dihydrofolate reductase (DHFR) or Schizosaccharomyces pombe TPI gene, or ampicillin, kanamycin, tetracycline, chloramphenicol, Mention may be made of drug resistance genes such as neomycin or hygromycin. A method for linking a wild-type CCN1 gene or a modified CCN1 gene, a promoter, and a terminator and / or a secretion signal sequence, if necessary, and inserting them into an appropriate vector is well known to those skilled in the art.

  A transformant can be prepared by introducing the expression vector into a suitable host, and the transformant can be cultured to prepare a wild-type CCN1 protein or a modified CCN1 protein. The type of host cell is not particularly limited as long as it can express the gene. For example, bacteria (eg, Escherichia, Bacillus, etc.), yeast (Saccharomyces, Pichia, Candida, etc.), animal cells (CHO cells, COS cells, etc.), insect cells (Sf-9 cells, Hi-five cells) Etc.) can be used. A liquid medium is suitable as a medium for culturing, and it is particularly preferable that the medium contains a carbon source, a nitrogen source and the like necessary for the growth of the transformed cells to be cultured. If desired, vitamins, growth promoting factors, serum and the like can be added. As a method for introducing an expression vector into a host cell, for example, an appropriate method such as an electroporation method, a calcium phosphate method, or a lipofection method can be used.

  The obtained wild-type CCN1 protein or modified CCN1 protein can be purified according to a conventional method and used as an active ingredient of the medicament of the present invention. Examples of purification means include solvent extraction, salting out using ammonium sulfate, desalting, precipitation using organic solvents, anion exchange chromatography using a resin such as diethylaminoethyl (DEAE) sepharose, S-Sepharose Cation exchange chromatography using a resin such as FF (Pharmacia), hydrophobic chromatography using a resin such as butyl sepharose and phenyl sepharose, gel filtration using a molecular sieve, one affinity chromatography, chromatography Techniques such as focusing methods and electrophoresis methods such as isoelectric focusing can be used alone or in combination, but are not limited thereto.

The medicament of the present invention containing the wild-type CCN1 protein or the modified CCN1 protein as an active ingredient can be formulated according to a general method for preparing a protein preparation. The medicament of the present invention containing a wild-type CCN1 gene or a modified CCN1 gene or an expression vector incorporating them as an active ingredient can also be used after being formulated by means available in the art. Among the above-described expression vectors, the medicament of the present invention containing a wild-type CCN1 gene or a modified CCN1 gene can efficiently express a wild-type CCN1 protein or a modified CCN1 protein in vivo in mammals including humans. Vectors can be used. It is preferable to incorporate into this vector sequences necessary for gene expression and various sequences that promote gene expression in an appropriate order. Various gene therapies for expressing a target gene in a living body, preferably a specific organ or tissue, are performed in the art, and the pharmaceutical of the present invention containing the above expression vector is used according to the gene therapy method. Thus, CCN1 protein or modified CCN1 protein can be expressed in vivo, preferably in the heart, and efficient gene therapy can be performed.

  The medicament of the present invention has an action of protecting cardiomyocytes from oxidative stress and suppressing apoptosis. In addition, the medicament of the present invention exhibits an angiogenic action in the heart and has an action of improving cardiac function. Therefore, the medicament of the present invention can be used as a medicament for the prevention and / or treatment of various heart diseases. A particularly preferable application target is a heart disease in which a strong oxidative stress is applied to cardiomyocytes, and more specifically, an ischemic heart disease such as angina pectoris and acute myocardial infarction. In addition, oxidative stress during the resumption of blood flow during treatment using balloon catheters or stents such as percutaneous coronary angioplasty (PTCA) or percutaneous coronary intervention (PCI) performed for acute myocardial infarction The drug of the present invention is preferably used for alleviation, prevention of restenosis that frequently occurs after PTCA or PCI treatment, and reduction of oxidative stress during reperfusion during coronary aortic bypass surgery (CABG or OPCAB) Can do. Furthermore, since the medicament of the present invention has an angiogenic action and a cardiac function improving action in the heart, cardiomyopathy (hypertrophic cardiomyopathy, secondary myocardial hypertrophy, dilated cardiomyopathy, restricted cardiomyopathy, etc.) , Myocarditis, heart failure, endocarditis (such as bacterial endocarditis), valvular disease (such as mitral regurgitation), pericarditis (such as acute pericarditis, chronic constrictive pericarditis), The medicament of the present invention can be used for prevention and / or treatment of heart diseases such as congenital heart diseases (eg, atrial septal defect, ventricular septal defect), cardiac asthma, and pulmonary heart. But the application object of the medicine of this invention is not limited to said disease.

  The method of applying the medicament of the present invention is not particularly limited, but generally, parenteral systemic administration such as intravenous administration, subcutaneous administration, intramuscular administration, inhalation, or local administration by injection into the heart, etc. should be adopted. Can do. Needless to say, the route of administration can be appropriately selected according to the type of active ingredient, the type and symptoms of heart disease, and the like. In general, the medicament of the present invention can be prepared as a pharmaceutical composition in a parenteral dosage form using one or more pharmaceutical additives. The kind of additive for formulation is not specifically limited, Generally, the additive for formulation used for preparation of a protein formulation or a nucleic acid formulation can be selected, and it can use combining suitably as needed.

Preferable pharmaceutical forms include, but are not limited to, injections or infusions that can be prepared at the time of use in a lyophilized state, or intravenous injection solutions or infusions. Examples of pharmaceutical additives for manufacturing injections include pH adjusters, buffers, isotonic agents, solubilizers, soothing agents, stabilizers, preservatives, or antioxidants. In addition to the above-mentioned additives for preparation, excipients and the like can be used for the production of a lyophilized form of the medicament, but it is not limited thereto. A coating containing the active ingredient of the medicament of the present invention may be applied to a balloon catheter or an indwelling stent so that the active ingredient is gradually released in the coronary artery, or the medicament of the present invention is applied to the heart during thoracotomy. It can also be applied directly. Also, any drug delivery system can be used so that the medicament of the present invention is specifically accumulated in the heart.
The dosage of the medicament of the present invention is not particularly limited and can be appropriately selected depending on the type of heart disease, the purpose of prevention or treatment, the age, symptoms, body weight, sex, etc. of the patient, but generally 1 mg per day About 20mg.

Hereinafter, the present invention will be described more specifically with reference to examples. However, the scope of the present invention is not limited to the following examples.
Example 1: Cardiomyocyte protective effect of CCN1 in cultured rat cardiomyocytes Oxidative stress due to the production of active oxygen is an important factor causing myocardial damage during reperfusion injury in acute myocardial infarction and remodeling in chronic heart failure Therefore, protecting cardiomyocytes from oxidative stress is important from the viewpoint of maintaining and improving cardiac function. Therefore, the effect of CCN1 on cardiomyocyte protection from oxidative stress was examined. The CCN1 recombinant protein used in the following experiment was prepared by the method described in the revised 3rd edition protein experiment note (Yodosha, 2004) and the like.

Primary cultured cardiomyocytes were prepared by treating collagenase and trypsin on the heart of newborn rats 2 to 3 days after birth. After culturing in a medium containing serum for 24 hours and further in a medium not containing serum, 2 μg / ml CCN1 or PBS was added and cultured for 24 hours. To these, 200 mM H ₂ O ₂ was added for oxidative stress, and 24 hours later, live cells and dead cells were dyed using calcein and PI to count the ratio of live cells. The results are shown in FIG. The survival rate of cardiomyocytes was 21% in the control (PBS) but 69% in the cardiomyocytes added with 2 μg / ml CCN1, and CCN1 significantly increased the survival rate of cardiomyocytes under oxidative stress It was shown that. In addition, 24 hours after adding H ₂ O ₂ , TUNEL staining was performed to count the number of cells undergoing apoptosis. The results are shown in FIG. TUNEL-positive cells were 42% in the control and 23% with the addition of CCN1, indicating that CCN1 significantly suppressed cardiomyocyte apoptosis due to oxidative stress.

  Furthermore, cardiomyocyte signals induced by CCN1 were analyzed by Western blotting. After isolation of cardiomyocytes, culture for 24 hours in serum-containing medium and 24 hours in serum-free medium, then add 2 μg / ml CCN1 or PBS and dissolve cells to separate proteins 30 and 60 minutes later The amounts of Akt, ERK, phosphorylated Akt, and phosphorylated ERK were compared by Western blot. The results are shown in FIG. CCN1 was found to promote phosphorylation of Akt and ERK in cardiomyocytes.

Therefore, in order to investigate whether the survival of cardiomyocytes under oxidative stress by CCN1 is transmitted through these signals, together with CCN1, PI3K-Akt inhibitor wortmannin, MEK-ERK inhibitor PD098059 In addition, the effect of CCN1 on the survival effect was examined. After isolation of cardiomyocytes, culture for 24 hours in medium containing serum, and further for 24 hours in medium without serum, followed by 2 μg / ml CCN1, CCN1 + 100 μM wortmanin (inhibitor of PI3K-Akt), CCN1 + 30 μM PD98059 (MEK-ERK Inhibitors) or PBS, culture for 24 hours, add 200 mM H ₂ O ₂ to apply oxidative stress, and after 24 hours, live and dead cells are dyed separately using calcein and PI. The cell ratio was counted. As a result, it was revealed that the cardiomyocyte survival effect of CCN1 is exerted through both Akt and ERK, particularly Akt (FIG. 4).
From these experimental results, it was shown that CCN1 is a protein that has both angiogenic activity and cardiomyocyte survival effect under oxidative stress.

Example 2: Cardiac function improvement effect of myocardial infarction model of CCN1
In order to examine the effect of CCN1 in ischemic heart disease, an experiment using a rat myocardial infarction model was performed. After ligating the anterior descending branch of the rat and performing echocardiography 4 weeks later, we selected rats (20 of 50) that had myocardial infarction of the expected size and randomly selected them in 2 groups (CCN1 group and In the PBS group, the chest was restarted under general anesthesia, and a gelatin sheet containing 100 μg CCN1 was spread on the heart surface in the CCN1 group (10 mice), and the PBS group (10 mice) contained only PBS. Spread the gelatin sheet on the surface of the heart and complete the surgery. Four weeks after surgery, echocardiography and mirror catheter examination were performed under general anesthesia. After these tests, the animals were euthanized and the heart was removed for histological examination.

Echocardiography measured left ventricular diastolic diameter, left ventricular systolic diameter, ejection fraction, left ventricular septal thickness, and left ventricular free wall thickness from the chest wall. In the mirror catheter examination, a catheter was inserted from the rat carotid artery to the left ventricle, and various parameters were measured including dP / dt and -dP / dt, which are indices of cardiac contractility and dilatability. In the histological examination, the morphology was examined by H & E staining, and vascular endothelial cells were stained with an anti-von Willebrand factor antibody, and the capillary density at the site close to the infarct was measured. It was shown that the blood vessel density was significantly increased in the CCN1 administration group (FIG. 5), and it became clear that the administration of CCN1 promotes angiogenesis in the heart. Furthermore, in mirror catheter examinations, dP / dt and -dP / dt, which are indices of contraction function and dilation function, are significantly higher in the CCN1 administration group (Fig. 6), and the cardiac function is significantly improved by CCN1 administration compared to the control group. It was shown that.
These results indicate that CCN1 promotes cardiovascularization and further improves cardiac function in ischemic heart disease.

It is the figure which showed the survival rate of the cardiomyocyte in CCN1 presence. It is the figure which showed that the apoptosis of the cardiac muscle cell was suppressed by CCN1. It is the figure which showed the result of having analyzed the signal in a cardiac muscle cell induced | guided | derived by CCN1 by Western blotting. It is the figure which showed that the cardiomyocyte protective effect of CCN1 is mainly via the activation of Akt. It is the figure which showed that angiogenesis is accelerated | stimulated in an ischemic heart by CCN1. It is the figure which showed the cardiac function improvement effect of CCN1.

Claims (7)

A medicament for the prevention and / or treatment of heart disease comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient. The above medicament, wherein the heart disease is ischemic heart disease, heart failure or cardiomyopathy. The above medicament, wherein the ischemic heart disease is acute myocardial infarction or angina. A cardiac function improving agent comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient. An angiogenesis promoter for heart comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient. An agent for protecting a heart from oxidative stress, comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient. An inhibitor of apoptosis of heart caused by oxidative stress, comprising CCN1 protein or a gene encoding CCN1 protein as an active ingredient.