KR20050031499A - Composition for preventing and treating brain ischemic disease - Google Patents

Abstract

A composition for preventing and treating cerebral ischemic diseases is provided, which composition inhibits apoptosis of cells in the hippocampus region caused by cerebral ischemia-reperfusion. The composition for preventing and treating cerebral ischemic diseases comprises at least one compound selected from 4-hydroxybenzyl alcohol, of formula (1), 4-hydroxybenzyl aldehyde of formula (2) and vanillin of formula (3), wherein the cerebral ischemic disease is selected from cerebrovascular accident, cerebral infarction, intracerebral hemorrhage, epilepsy and Parkinson's disease.

Description

Composition for preventing and treating brain ischemic disease

The present invention relates to a composition for the prevention and treatment of cerebral ischemic disease, and more particularly, to a composition for the prevention and treatment of cerebral ischemic disease that effectively inhibits the damage of nerve cells caused by cerebral ischemia and reperfusion.

Cerebral ischemic disease is a disease caused by delayed neuronal death in the hippocampus CA1 region due to decreased blood flow in the brain and lack of oxygen and glucose (Kirino, Brain Res. , 239: 57-). 69, 1982; Petito et al., Neurology , 37: 1281-1286, 1987; Pulsinelli et al., Ann. Neurol. , 11: 491-498, 1982). In addition, the cerebral ischemic brain disease is known to be caused by the lack of blood supply to the brain in pathological conditions such as stroke or heart attack (Nedergaard, Acta. Neurol . Scand. , 77: 81-101, 1988; White; et al., Neurology , 43: 1656-1665, 1993).

The cause of delayed neuronal death in the hippocampal CA1 region caused by transient cerebral ischemia and reperfusion is caused by the influx of calcium into cells (Hara et al., Brain Res. Bull ., 29: 659-665, 1992; Nedergaard , Acta. Neurol. Scand. , 77: 81-101, 1988), known as free radical related neuronal damage and glutamate-receptor mediated neuronal damage (Won et al., Neurosci. Lett. , 301: 139-142, 2001). That is, when cerebral ischemia occurs, depolarization of neurons occurs, and glutamate of synapses is released, thereby increasing the concentration of extracellular glutamate. In addition, the reversal of active transport resulting from a decrease in ATP accelerates the accumulation of extracellular glutamate. Accumulation of the extracellular glutamate may include N-methyl-D-aspartic acid (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), and metabotropic glutamate receptor. ), Resulting in an over-flow of calcium into cells (Olney et al., Science , 244: 1360-1362, 1989). Intracellular influx of calcium triggers the activity of calcium-dependent proteases, lipases and modulators, and eventually results in the generation of cytotoxic molecules including free radicals, which destroy cell structures such as DNA and membranes, resulting in neuronal death. Known.

Treatments for cerebral ischemic diseases currently being used in the clinic include thrombolytics such as tissue plasminogen activator (TPA) and urokinase, ticlopidine, cilostazole and prostacycline. Drugs of various mechanisms include platelet inhibitors, antithrombin agents, anticoagulants, cerebrovascular agents, Ca ²⁺ -channel blockers, and brain edema inhibitors (Sandercock et al., Hosp. Med., 47: 731-736, 1992). However, these agents are known to have a weak effect when treatment is delayed, do not effectively block the progression of cerebral infarction due to acute cerebral ischemia, and have side effects such as nonspecific bleeding, fibrinogen lysis, and acute reoccurrence.

Recently, many researchers have attempted a multi-faceted approach to develop new therapeutics that can block or suppress irreversible damage based on the mechanism of neuronal damage caused by cerebral ischemia. In other words, glutamate receptor antagonists such as NMDA and AMPA, Ca ²⁺ -channel blockers, Na ⁺ -channel blockers, free radical scavengers, calpain inhibitors, nitric oxide synthase inhibitors, etc. Leeson PD and Iversen LL, J. Med. Chem ., 37: 4053-4067, 1994; Muir KW and Lees KR, Stroke, 26: 503-513, 1995). As a result, several drugs are currently in the clinical trial stage, but the mechanism of neuronal damage is very complicated, and due to various problems such as side effects of drugs and penetration into brain tissue, effective treatments have not been developed. . Therefore, there is an increasing demand for a new type of cerebral ischemic disease therapeutic agent that can supplement the problems of existing therapeutic agents without side effects.

Meanwhile, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are all DPPH (1,1-diphenyl-2-picryl hydrazyl) radicals, It is known to have an antioxidant effect that effectively removes superoxide radicals and hydroxyl radicals (Liu J. and Mori A., Neuropharmacology , 31: 1287-1298, 1992; Luo H et. al., Hua Hsi I Ko Ta Hwueh pao , 23: 53-56, 1992).

However, it is not yet known that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are effective in the prevention and treatment of cerebral ischemic disease.

The inventors of the present invention are excellent in the treatment of cerebral ischemic disease, and while studying to find a substance having no side effects, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin are effective in the treatment and prevention of cerebral ischemic disease. The present invention was completed by confirming that it can be used as a therapeutic agent for cerebral ischemic disease.

Accordingly, it is an object of the present invention to provide a composition for the prevention and treatment of cerebral ischemic disease containing at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin.

In order to achieve the above object, the present invention comprises at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol of formula (1), 4-hydroxybenzyl aldehyde of formula (2) and vanillin of formula (3) Provided is a composition for preventing and treating cerebral ischemic disease.

<Formula 1>

<Formula 2>

<Formula 3>

Hereinafter, the present invention will be described in detail.

The present invention is characterized by providing a composition for preventing and treating cerebral ischemic disease containing at least one compound selected from the group consisting of 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin.

The 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin may be purchased commercially or prepared by chemical synthesis methods known in the art. It is also possible to separate and purify from natural, preferably plants, and particularly preferably chick horses . As a method for separating and purifying compounds according to the present invention from cheonma , a method known in the art may be used (Kiyoshi Hata et al., Yakugaku Zasshi ., 83: 606-610, 1963; Kuk Hyun Shin et al., Arch Pharm.Res ., 17: 331-336, 1994). Preferably, a pure compound of interest can be obtained by solvent extraction of the active ingredient of cheonma and then separation and purification by chromatography.

In one embodiment of the present invention, to determine whether the 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be used as a therapeutic agent for cerebral ischemic disease, the common carotid arteries are closed for 5 minutes. A transient whole cerebral ischemia model was used to investigate whether the compounds effectively inhibit neuronal death caused by ischemia. To this end, the experimental animals were divided into the manipulator group, the control group, the 4-hydroxybenzyl alcohol-administered group, the 4-hydroxybenzyl aldehyde-administered group, and the vanillin-administered group. It was.

As a result, almost no neuronal damage was found in the singular group (100% viable cells), whereas severe cell damage was observed in the control group (11.7% viable cells) (see FIG. 1). On the other hand, when administering the compounds according to the present invention, there was a slight difference depending on the type of compound, the sex of the animal and the time of administration, the number of viable cells was significantly higher than the control (see Fig. 2a to 4b). The number of surviving cells in the groups administered with the compound according to the present invention was higher in females than in males, and the administrations before the ischemic induction were higher than those after the ischemic induction (except in the vanillin female administration group), and approximately 4-hydroxybenzyl. Alcohol (average 52.80%), vanillin (average 52.13%), and 4-hydroxybenzyl aldehyde (average 32.55%) were the highest (see FIG. 5).

The above results indicate that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can effectively inhibit hippocampal neuronal cell death caused by cerebral ischemia, and thus have an excellent effect in preventing or treating cerebral ischemic disease. Indicates that there is.

In another embodiment of the present invention, in order to confirm the mechanism of action of the treatment effect of cerebral ischemic disease of 4-hydroxybenzyl alcohol showed the best effect of inhibiting neuronal cell damage caused by cerebral ischemia, the time after ischemic induction The changes of NMDA receptor type 1, 8-hydroxy-2'-deoxyguanosine and GABA transaminase, respectively, were investigated. It was. The NMDA receptor type 1 is known to be involved in the intracellular influx of calcium, and 8-hydroxy-2'-dioxyguanosine appears when DNA is damaged by oxidative stress, which is caused by the oxidation of guanosine by radicals. It is known to produce the substance, GABA transaminase is known to promote the amino transfer reaction of GABA.

As a result, when 4-hydroxybenzyl alcohol was administered intraperitoneally 30 minutes before the induction of cerebral ischemia, the expression level of NMDA receptor type 1 was not changed compared with the case without administration (see FIG. 6), and 8-hydroxy-2 The immune response of '-deoxyguanosine was slightly reduced (see FIG. 7), and the amount of GABA transaminase increased rapidly (see FIG. 8).

The above results show that 4-hydroxybenzyl alcohol according to the present invention has little effect on the inhibition of calcium influx generated during ischemia, but inhibits DNA damage caused by oxidative stress and reduces the energy generated by fast GABA degradation. It is thought to reduce cell damage caused by cerebral ischemia by supplying pyramidal neuron cells in the region to help cell survival. On the other hand, the mechanism of treatment of cerebral ischemic diseases of 4-hydroxybenzyl aldehyde and vanillin according to the present invention is thought to be able to inhibit cell death by preventing excessive excitability of neurons by inhibiting GABA transaminase (result not shown). And the mechanism of treatment of the 4-hydroxybenzyl alcohol.

4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be administered to a patient after being formulated in a known manner by further comprising a pharmaceutically acceptable carrier. The term 'pharmaceutically acceptable' refers to a composition which is physiologically acceptable and does not cause an allergic or similar reaction, such as gastrointestinal disorders, dizziness or the like, when administered to humans.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. Other pharmaceutically acceptable carriers may be referred to those described in the following documents (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin according to the present invention can be prepared in various parenteral or oral administration forms according to known methods by mixing with a pharmaceutically acceptable carrier as described above. have. For example, typical of parenteral formulations are injectable formulations, preferably aqueous isotonic solutions or suspensions. Injectable formulations may be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component may be formulated for injection by dissolving in saline or buffer. In addition, oral dosage forms include, for example, powders, granules, tablets, pills, capsules, and the like, which include, in addition to the active ingredient, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and the like). / Or glycine), a lubricant, such as silica, talc, stearic acid and its magnesium or calcium salt and / or polyethylene glycol. Tablets may also include binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethyl cellulose and / or polyvinylpyrrolidine, optionally starch, agar, alginic acid or its Disintegrants or boiling mixtures such as sodium salts and / or absorbents, colorants, flavors, and sweeteners. The formulations may be prepared by conventional mixing, granulating or coating methods.

Pharmaceutical compositions formulated in such a manner can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

As used herein, the term 'effective amount' refers to an amount of a compound that exhibits an effect of preventing or treating cerebral ischemic disease when administered to a patient. Preferably a typical daily dosage of the compounds according to the invention is in the range of 40 to 60 mg / kg body weight. The compounds according to the invention can be administered once or in divided doses within the preferred dosage range. However, the dosage of the compounds according to the present invention may be appropriately selected depending on the route of administration, subject to administration, age, gender weight, individual difference and disease state. As used herein, a patient refers to a mammal including a human, and preferably a human diagnosed with cerebral ischemic disease. The cerebral ischemic diseases include all diseases caused by cerebral ischemia and reperfusion. For example, but not limited to, stroke, cerebral infarction, cerebral hemorrhage, epilepsy or Parkinson's disease, and the like.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited to the following examples.

<Example 1>

Confirmation of the pharmacological effect of the compound according to the present invention

1-1) Induction of Temporary Whole Cerebral Ischemia Injury

The experimental animal was a Mongolian gerbil, weighing 65-75 g, which is a descendant of the Mongolian gerbil ( Meriones unguiculatus ), which was distributed from the Hallym University Experimental Animal Center. They were kept for 14 days at 23 ± 2 ° C. and 50 ± 10% relative humidity under constant light and light cycles until 7 pm.

In order to cause transient cerebral ischemia, a gas mixture consisting of 67% by volume of nitric oxide and 33% by volume of oxygen in addition to 2.5% by volume of isoflurane (Baxtor, USA) was added. General anesthesia was performed by a general anesthesia method, followed by hair cutting and disinfection of the neck of the Mongolian gerbil. The neck was then incised to expose both common carotid arteries and ligation was induced for 5 minutes using a non-traumatic aneurysm clip (US Staelting). During induction of ischemia, complete obstruction of the total carotid artery was confirmed by observing the blood circulation of the central artery of retina using an ophthalmoscope. The rectal thermometer was inserted to measure body temperature. By using a thermal pad that is automatically adjusted according to the temperature of the animal body temperature was kept constant at 37 ± 0.3 ℃ normal temperature. After the ischemia induction, the aneurysm clip was removed and reperfused, and the reperfusion was observed directly under a microscope.

4-hydroxy was suspended in 1 ml of 10% Tween 80 (dissolved in 0.9% saline solution) 30 minutes before ischemia and 30 minutes after ischemia induction for female and male Mongolian gerbils induced by ischemia. 4-hydroxybenzyl alcohol administration group, benzyl alcohol (Fluka Co.), 4-hydroxybenzyl aldehyde (Junsei Chemical Co.) and vanillin (Sigma Chemical Co.) intraperitoneally administered in an amount of 40 mg / kg body weight, respectively, 4 The hydroxybenzyl aldehyde administration group and the vanillin administration group were prepared. At this time, the group administered with the 10% Tween 80 solution dissolved in the same amount of 0.9% physiological saline 30 minutes before ischemic induction and 30 minutes after tongue induction was used as a control. For another group of Mongolian gerbils, all procedures were performed on the same sham procedure except that the carotid artery was not ligated and caused ischemia (surgery group). Animal groups treated differently as above are summarized in Table 1 below.

Singer Control 4-hydroxybenzyl alcohol administration group 4-hydroxybenzyl aldehyde administration group Vanillin-administered group cock 30 minutes before ischemia induction 7 7 7 7 7 30 minutes after ischemic induction 7 7 7 7 7 female 30 minutes before ischemia induction 7 7 7 7 7 30 minutes after ischemic induction 7 7 7 7 7

1-2) Observation and measurement of nerve cell damage

The animal group was sacrificed 4 days after the day of cerebral ischemia-induced surgery. All Mongolian gerbils were intraperitoneally injected with thiopental sodium (Yuhan, Korea) at a dose of 40 mg / kg body weight, anesthetized and left ventricular at 4 ° C. containing 1000 IU of heparin per 1,000 ml. Perfusion washed by injection. After perfusion washing, the animals were perfused with 4% paraformaldehyde dissolved in 0.1 M phosphate buffer (PB, pH 7.4).

After perfusion fixation, the brain was removed by using a bone cutter to extract the brain, and then fixed in the same fixative after 4-6 hours. The post-fixed brains were soaked in 30% sucrose solution dissolved in 0.1 M phosphate buffer and soaked until they sank to the bottom. Thereafter, using a sliding microtome (sliding microtome, Reichert-Jung, Germany) was cut into 30 ㎛ thickness into a 6 well plate containing a storage solution (storing solution) and stored at 4 ℃. Choose a tissue that shows well hippocampal formation among each tissue section, remove the preservatives from the tissue, and dry it on a gelatinized slide. Thereafter, the solution was briefly dipped in distilled water, dyed in 2% cresyl violet (Sigma, USA) solution for 1 minute, and then washed in running tap water to remove excess cresyl violet on the slide.

Hippocampal CA1 damage was determined by counting viable neurons stained with cresyl violet solution. For each group of animals, the average number of CA1 neurons per millimeter of bilateral hemispheres in hippocampal sections was measured. The measurement of the number of neuronal somata was performed using an image analysis system (software: Optimas 6.5, CyberMetrics, USA) equipped with a computer-based CCD camera. Cells were counted by averaging the number of stained neurons from 21 sections obtained from each animal. All data obtained from quantitative data to verify statistical significance were analyzed using the One-way ANOVA test and Bonferroni's test was performed for post hoc comparison. It was. Significance was determined when p value was 0.01 or less.

Micrographs of the hippocampal tissue and its CA1 region of each group stained with cresyl violet are shown in FIGS. 1 to 4. As shown in FIG. 1, almost no cell damage was found in the ischaic group that did not induce ischemia (FIGS. 1A and 1C), whereas there was severe cell damage in the control group (FIGS. 1B and 1D). Meanwhile, the 4-hydroxybenzyl alcohol-administered group, 4-hydroxybenzyl aldehyde-administered group, and vanillin-administered group showed more severe cell damage than the hydrolyzate group, but significantly reduced cell damage compared to the control group (FIGS. 2A to 2). 4b).

5 shows the results of counting the number of viable cells in each group based on the manipulator group (100%) having almost no neuronal damage. As shown in FIG. 5, the number of surviving cells in the groups administered with the compound according to the present invention was higher in females than in males, and more frequently administered before ischemic induction than when administered after ischemic induction (except vanillin-administered group). 4-hydroxybenzyl alcohol (average 52.80%), vanillin (average 52.13%) and 4-hydroxybenzyl aldehyde (average 32.55%).

As a result, it was confirmed that 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin effectively inhibit the cell death of hippocampal region cells caused by cerebral ischemia and reperfusion.

<Example 2>

Confirmation of functional mechanism of 4-hydroxybenzyl alcohol

The inventors of the present invention to determine the mechanism of action of 4-hydroxybenzyl alcohol shown to most effectively inhibit the cell death of hippocampal region cells caused by cerebral ischemia and reperfusion among the compounds of the present invention, after ischemic induction Changes in NMDA receptor type 1, 8-hydroxy-2'-deoxyguanosine and GABA transaminase, respectively, over time Investigate.

A cerebral ischemia-induced experimental animal was prepared in the same manner as in Example 1. 40 mg / kg of 4-hydroxybenzyl alcohol was intraperitoneally administered to Mongolian gerbil 30 minutes before the cerebral ischemia was induced. Thereafter, animals were sacrificed at 30 minutes, 3 hours, 6 hours, 12 hours, and 24 hours before ischemia induction, respectively, and the perfusion fixation was performed in the same manner as in the above example, and the tissue was 30 μm thick. Cut and stored in 6 well play containing the stock solution. Choose a tissue that shows well hippocampus among each tissue section, wash it three times for 10 minutes with 0.1M PBS (phosphate buffered saline, pH 7.4) to remove the preservative solution from the tissue, and then perform immunostaining using the following ABC method. It was. That is, the washed tissue sections were treated with 0.3% H ₂ O ₂ in 0.01 M PBS for 30 minutes to remove endogenous peroxidase present in the tissue, and 5% to prevent nonspecific reactions. Standard goat serum was treated and reacted for 30 minutes. The primary antibodies were mouse anti-NMDA receptor type 1 (chemicon International, USA) and mouse anti-8-hydroxy-2'-deoxyguanosine (mouse anti-8-hydroxy-) 2'-deoxyguanosine: Peninsula, USA and mouse anti-GABA transaminase (provided by Professor Soo Young Choi, Hallym University) contain 2% standard goat serum and 0.1% Triton X-100 (Triton X-100) The diluted 0.1 M PBS solution was diluted at a ratio of 1: 100 and reacted with each of the treated tissues at 4 ° C. for 24 hours. Afterwards, the biotinylated goat-mouse IgG (Biotinylated-goat Anti-mouse IgG: Vector of USA) was used as a secondary antibody and 0.1M containing 2% standard goat serum and 0.1% Triton X-100. The solution was diluted 1: 200 in PBS solution and reacted with each of the tissues at room temperature for 2 hours, after which the tissue was reacted with the vector kit (US Vector) solution at room temperature for 1 hour. After each reaction, the tissues were washed four times with PBS solution. After the antigen-antibody reaction, the tissue was developed for 1-2 minutes in a solution of Tris buffer (pH 7.4) containing 0.03% hydrogen peroxide solution and 0.05% DAB (3,3'-diaminobenzidine tetrachloride, Sigma, USA). After washing, it was dried on gelatin-coated slides, dried, and then sealed with Canadian balsam (Kanto, Japan) through a conventional dehydration and clarification process. The changes of NMDA receptor type 1, 8-hydroxy-2'-dioxyguanosine and GABA transaminase with time after ischemic induction through the immunohistochemical staining are shown in FIGS. 6 to 8, respectively.

As shown in FIG. 6, the expression of NMDA receptor type 1, which is known to be involved in the intracellular influx of calcium during ischemia development, has a slight change over time after cerebral ischemia induction (FIG. 6). The results are consistent with previous changes of the same experiments by the present inventors except that 4-hydroxybenzyl alcohol was administered prior to ischemic induction (Kang et al., J Neurocytol ., 30: 945-955, 2001) . will be. The results showed that 4-hydroxybenzyl alcohol did not affect the expression of NMDA receptor type 1, so 4-hydroxybenzyl alcohol had little effect on the influx of early calcium during ischemia. .

On the other hand, as shown in Figure 7, 8-hydroxy-2'-dioxyguanosine, an oxide produced by DNA damage due to oxidative stress (oxidative stress) slightly increased up to 12 hours to induce cerebral ischemia, The immune response at 12 hours of cerebral ischemia was increased by 150% compared to before ischemia induction. The above results indicate that the previous results showed that the immune response increased more than 200% in the same experiments performed by the inventors except that 4-hydroxybenzyl alcohol was administered before ischemic induction (Won et al., Brain Res., 836: 70-78, 1999). The results showed that administration of 4-hydroxybenzyl alcohol inhibited the increase of 8-hydroxy-2'-dioxyguanosine, thus inhibiting DNA damage due to oxidative stress occurring early in ischemia development. I think.

On the other hand, as shown in Figure 8, in the case of GABA transaminase to promote the aminotransferase reaction of GABA, it began to increase rapidly 3 hours after ischemia induction. The results indicate that the rapid increase of GABA transaminase occurred 12 hours after ischemic induction in the experiments performed by the inventors in the same manner except that 4-hydroxybenzyl alcohol was administered before ischemic induction (Kang et al. , Neurosci Lett., 294: 33-36, 2000). As a result, administration of 4-hydroxybenzyl alcohol lowered the ability to inhibit the increase of GABA transaminase, and thus the energy generated by fast GABA degradation was reduced to pyramidal cells in the CA1 region. It is thought to contribute to the survival of cells by supplying them to neuron cells.

As described above, 4-hydroxybenzyl alcohol, 4-hydroxybenzyl aldehyde and vanillin have the effect of inhibiting cell death of hippocampal site cells caused by cerebral ischemia-reperfusion. Thus, the compounds can be effectively used for the prevention and treatment of cerebral ischemic disease.

Figure 1 shows a micrograph of the hippocampal tissue and its CA1 region stained with cresyl violet (A: hippocampus tissue of the Singer group, B: hippocampal tissue of the control group, C: Singer surgery group) Hippocampus CA1 region, D: hippocampus CA1 region of the control group).

2A and 2B show microscopic photographs of the hippocampal tissue and its CA1 region of the 4-hydroxybenzyl alcohol-administered group with cresyl violet, respectively (A: 30 minutes before ischemia-induced group (male), B: ischemia-induced 30). Post-administration group (male), C: administration group 30 minutes before ischemia-induced (female), D: administration group (female) 30 minutes after ischemia-induced).

3A and 3B show micrographs of the hippocampal tissue of the 4-hydroxybenzyl aldehyde-administered group and its CA1 region stained with cresyl violet (A: 30 minutes before ischemia-induced group (male), B: 30 minutes after ischemia-induced group) Administration group (male), C: administration group 30 minutes before ischemic induction (female), D: administration group (female) 30 minutes after ischemic induction).

4A and 4B show microscopic photographs of the hippocampus tissue and its CA1 region stained with cresyl violet, respectively (A: 30 minutes before ischemia-induced group (male), B: 30 minutes after ischemia-induced group (male)). C: group administered 30 minutes before ischemic induction (female), D: group administered 30 minutes after ischemic induction (female).

Figure 5 shows the average number of viable hippocampal cells after ischemic induction in the hippocampus tissue of each group based on the manipulator group.

Figure 6 is a micrograph of the hippocampus CA1 region by immunohistostaining for NMDA receptor type 1 when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction (A: cerebral ischemia, B). 30 minutes after cerebral ischemia, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).

FIG. 7 shows hippocampal CA1 due to immunohistostaining for 8-hydroxy-2'-deoxyguanosine when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction. A micrograph of the area (A: cerebral ischemia, B; 30 minutes after cerebral ischemia, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).

Figure 8 is a micrograph of the hippocampal CA1 region by immunohistostaining for GABA transaminase when 4-hydroxybenzyl alcohol was administered 30 minutes before ischemia induction (A: cerebral ischemia, B; cerebral ischemia). 30 minutes later, C: 3 hours after cerebral ischemia, D: 6 hours after cerebral ischemia, E: 12 hours after cerebral ischemia, F: 24 hours after cerebral ischemia).

Claims (2)

At least one compound selected from the group consisting of 4-hydroxybenzyl alcohol of formula 1, 4-hydroxybenzyl aldehyde of formula 2 and vanillin of formula 3 Composition for the prevention and treatment of cerebral ischemic disease containing. <Formula 1> <Formula 2> <Formula 3> The composition for preventing and treating cerebral ischemic disease according to claim 1, wherein the cerebral ischemic disease is selected from the group consisting of stroke, cerebral infarction, cerebral hemorrhage, epilepsy and Parkinson's disease.