WO2013004145A1 - Composition of chinese herbal compound or extract and use thereof - Google Patents

Abstract

Provided are a composition of a Chinese medicinal compound or extract and a use thereof. The Chinese medicine consists of ginseng, coptis, and gardenia. Also provided is a use of the composition in preparing a medicament for treating or preventing nerve injury, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression.

Description

 Composition of traditional Chinese medicine compound or extract and use thereof

Technical field

 The present invention relates to a composition of a traditional Chinese medicine compound or extract, wherein the traditional Chinese medicine consists of ginseng, berberine and medlar. The invention further relates to the use of the composition for the manufacture of a medicament for the treatment or prevention of brain and nerve damage, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression. Background technique

 Traditional Chinese medicine theory attaches importance to the overall concept and syndrome differentiation. Traditional Chinese medicine theory believes that qi deficiency and blood stasis is an important pathogenesis of stroke. Qi deficiency is the basis of this disease. Blood stasis is the basic pathogenesis of this disease. It is the core of the disease's development and development. The basic method of treating this disease. Qi deficiency and blood stasis are only seen in the traditional sequelae of sequelae. Yiqihuoxue method is only used for sequelae patients. Qing Dynasty famous doctor Wang Qingren Bu Yang Huan Wu Tang is the representative prescription (Zhang Meikui, Yin Ling, Zhang Xuewen Bu Yang Huan Wu Tang) Research progress in the treatment of ischemic stroke, Chinese Journal of Basic Medicine in Traditional Chinese Medicine, 2002, 8 (9): 74-78). The use of Yiqi, detoxification and related Chinese medicine to treat brain and nerve damage diseases has not been reported.

 Senile dementia (SD), also known as Alzheimer disease (Alzheimer disease,

AD) is a central nervous system degenerative disease characterized by progressive cognitive impairment and memory impairment, with varying degrees of abnormalities in sports, cognition, language, and personality. The clinical manifestation of AD is dementia syndrome, characterized by memory loss and personality degeneration. The main pathological changes in AD are cerebral cortical atrophy, loss of nerve cells and synapses, neurofibrillary tangles, extracellular senile plaques, and cerebral vascular amyloidosis. There is currently no fundamental treatment for Alzheimer's disease. The current molecular mechanisms and related botanicals for the treatment of senile dementia are: 1) acetylcholinesterase inhibitor scutellarin; 2) excitatory amino acid and peptide transmitter regulators, such as auxin; 3) interference with beta-starch Protein (Αβ) fiber formation and deposition, such as ginkgo flavonoids, salvianolic acid, emodin, timosaponin and so on. 4) Antioxidant and free radical scavenging drugs, Salvia miltiorrhiza water extract salvianolic acid, tea extract tea polyphenols, Ginkgo biloba extract, ginsenoside, ginkgolides, etc.; 5) Improve microcirculation, such as tanshinone and danshensu ; 6) Calcium antagonists, such as apigenin.

Vascular Dementia (VD) is a general term for dementia syndrome caused by brain damage caused by a series of cerebrovascular factors. It is mainly caused by memory and cognitive dysfunction on the basis of cerebrovascular disease, or A disease associated with acquired intelligence, continuous impairment of language, spatial skills, and emotional or personality disorders. Clinically, it is characterized by memory loss, calculation, judgment, orientation and other progressive mental and behavioral disorders. It is a common disease and frequently-occurring disease in the middle-aged and elderly population. The exact pathological mechanism of VD is not fully understood. It is generally believed that cerebrovascular disease causes local brain tissue inflammation, metabolic disorders and oxidative stress loss, damages brain tissue of sufficient capacity, and seriously impairs advanced neurological functions such as memory and cognition. Especially when the lesion involves the frontal lobe, temporal lobe and limbic system, the symptoms of dementia are more likely to occur. Traditional Chinese medicine for treating VD includes: Huanglian Jiedu Decoction, Danggui Shaoyao San, Dihuang Yinzi, Tianzhi Granule, Congsheng Capsule, Shenma Yizhi Capsule, Liuwei Dihuang Decoction, Jiawei Guipi Decoction, Buyang Huanwu Decoction and so on. And the use of Yiqi, detoxification and related Chinese medicine has not yet See the report.

 Parkinson disease (PD), also known as tremor paralysis, is a common central nervous system disease in middle-aged and elderly people. The clinical manifestations include static tremor, bradykinesia, myotonia and posture gait abnormalities. The pathogenesis is a dopaminergic (DA) neurological disorder in the central nigrostriatal pathway, leading to a loss of striatum DA, causing somatic motor dysfunction. When dopamine is reduced, acetylcholine is overexcited, and the two transmitters lose balance, causing nerves and muscle tissue to involuntarily produce symptoms such as rigidity and tremor. At present, there is no cure for Parkinson's disease. The western medicine levodopa (Medopa), bromocriptine, and ergocarpine are commonly used drugs in the clinic, but the effect is unstable, the side effects are large, and the long-term efficacy is not certain. Commonly used traditional Chinese medicine treatment methods and commonly used drugs are as follows: 1) Bugan deficiency, Zhengan Xifeng: Tianma, antler gum, asparagus, Ophiopogon japonicus, armor, turtle shell, raw white peony, licorice, etc.; 2) kidney deficiency Filling the qi and qi: 萸肉, Schisandra; 3) Complementing the blood, but also replenishing the brain: using Dayun, Angelica, longan meat, jujube, etc.; 4) Promoting blood circulation: Sanqi, pangolin, salvia, scorpion . Chen Jianzong et al. found that Chinese herbal medicines consisting of medlar, Cistanche, and Polygonum multiflorum can improve the content of DA, 3, 4-dihydroxyphenylacetic acid (DOPAC), NE, 5-HT in the striatum of PD rats. It has a certain protective effect on the nucleus of the substantia nigra, blue spot, and dorsal nucleus (Chen Jianzong, Guo Jianying, Sun Jing, etc., 40 cases of vascular Parkinson's syndrome treated by Peibu Ganshen Qufeng Tongluo Method, Journal of Traditional Chinese Medicine , 2002, 43(7): 528-529). He Jiancheng et al. found that Chinese herbal medicines with yin and tonifying kidney, collaterals and detoxification, which are composed of medlar, mulberry parasitic, tianma, uncaria, and silkworm, can significantly reduce the release of reactive oxygen species (ROS) and propylene from the brain of PD rats. The content of aldehyde (MDA) increases the content of free radical scavenging systems such as glutathione (GSH), glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD), which improves the body's antioxidant capacity. To alleviate the effects of free radicals on the body damage (He Jian, Yuan Canxing, Li Yaming, etc., the effect of nourishing liver, kidney, collaterals and detoxification Chinese medicine on oxidative stress in rat model of Parkinson's disease, Chinese Journal of New Drugs and Clinical Medicine, 2003, 22(3) ): 160-163). Xu Li and other observations of Xifeng Zhiyu Capsule (Danggui, Baiqi, Qi, Quanqi, etc., which are composed of Ziyin Yangxue and Xifeng Tongluo Traditional Chinese Medicine) for the treatment of experimental hypoxic experimental PD mice, which found that it can significantly increase PD The content of DA, DOPAC and high vanillic acid (HVA) in the brain of rats can reduce the intensity of limb tremor in mice and shorten the duration of tremor. Therefore, it is believed that Xifeng Zhiyu Capsule can prevent and treat PD (Xu Li, Wei Cuiyi, Liu Jianxun, etc. The preventive and therapeutic effects of Xifeng Zhizhi Capsule on experimental Parkinson's disease in mice, New Chinese Medicine and Clinical Pharmacology, 2005, 2: 87-90) The use of Yiqi, detoxification and related Chinese medicine has not been reported.

Depression usually refers to a kind of emotional disorder, mainly characterized by low mood, slow thinking, and reduced language action. It is mainly characterized by depression, decreased interest, pessimism, slow thinking, lack of initiative, and self-blame. Self-sin, diet, poor sleep, worry about having various diseases, feeling multiple discomforts in the body, serious suicidal thoughts and behaviors. Depression has the characteristics of high incidence, high prevalence, high recurrence rate, high suicide rate, low awareness rate and low treatment rate. The etiology of depression is complex. The pathological mechanisms hypothesis that has been studied are as follows: 1) Metabolic abnormalities of monoamine neurotransmitters, which are caused by insufficient monoamine transmitters in certain parts of the brain; 2) Changes in body function, antidepressants inhibit presynaptic monoamine neurotransmitter uptake is an immediate effect, while post-synaptic receptor sensitivity is a chronic adaptive change; 3) neuroendocrine disorders, studies found depression Hypothalamic-pituitary-adrenal axis (HPA), hypothalamic-pituitary-thyroid Functional disorders such as the glandular axis (HPT), hypothalamic-pituitary-growth hormone axis (HPGH), and decreased serum cholesterol levels are associated with decreased serum estrogen levels. At present, antidepressants mainly have three characteristics to varying degrees, namely, they have an stimulating effect on mental exercise, enhance internal drive, active emotions and anti-anxiety effects. It is mainly divided into five major categories: monoamine oxidase inhibitors, tricyclic antidepressants (TCAs, such as Datlan), tetracyclic antidepressants, new antidepressants such as selective 5-tryptamine reuptake inhibitors ( SSRIs, such as fluoxetine), selective norepinephrine reuptake inhibitors, serotonin and norepinephrine reuptake dual inhibitors (SNRIs such as venlafaxine), norepinephrine and specific 5- A tryptamine reuptake inhibitor (NaSSAs, such as mirtazapine), norepinephrine and dopamine reuptake inhibitors, botanical extracts, and the like. However, most of them have the characteristics of narrow anti-depression spectrum, large side effects, high drug price and easy recurrence.

 Meningitis can be divided into two categories: bacterial and sterilized. Bacterial meningitis is inflammation caused by bacterial infection of the pia mater. It is a serious intracranial infectious disease, often accompanied by encephalitis and brain abscess. From the perspective of pathogens, there are two major categories, one is meningitis of various purulent bacterial infections (abbreviation: brain), such as meningitis caused by meningococcal, pneumococcal, influenza, Haemophilus, etc. The other type is meningitis infected with Mycobacterium tuberculosis. Clinical medication is mainly antibiotics combined with hormone therapy. Aseptic meningitis refers to cerebrospinal fluid smears and bacterial cultures that are recessive meningitis. Generally, aseptic meningitis is caused by viral infection. Viral encephalitis refers to inflammation of the brain parenchyma caused by viral infections, often manifested as fever, headache, convulsions, disturbance of consciousness, and meningeal irritation, which can cause focal damage to the central nervous system. Viral encephalitis has a poor prognosis, high mortality, and often has serious sequelae. Commonly used drugs are 1) hormones, such as the adrenal cortex hormone dexamethasone; 2) antiviral steroids, such as acyclovir or adenosine; 3) antiviral cytokines, interferon; 4) nutrition Such as citicoline, vitamin B6, vitamin E, brain rehabilitation, pantothenic acid, etc. to improve brain metabolism, patients with seizures should also be combined with anti-epileptic drugs, patients with mental exercise excitement can use mentally stable drugs, etc. .

 There are still defects in the treatment of brain or nerve damage diseases, such as side effects, poor efficacy, and inability to cure. Because brain disease is a complex pathophysiological process involving multiple links, multiple factors, and multiple pathways, traditional Chinese medicine compound preparations can have multiple neuroprotective effects, with multi-target, multi-directional regulation, and clinical significance of prevention and treatment. It should be expected to be a more effective brain and neuroprotective drug.

 Various ginseng is a good medicine for tonic and is often used in Chinese medicine. "Shen Nong's Herbal Classic" records ginseng. "The main remedy for the five internal organs, the spirit of the spirit, the soul of the soul, the horror, the evil spirits, ..., "Happy Puzzle" is considered to be a kind of Chinese herbal medicine with various pharmacological effects. Its main active ingredients for brain and nerve damage have been reported in many cases, such as prevention and treatment of Alzheimer's disease, Parkinson's disease, depression, and cerebral infarction. Other diseases caused by cerebral ischemia and cerebral ischemia (Jiangshan, Jiang Zhenglin, Zeng Yinming, Research on the effects of nine kinds of ginsenosides against cerebral ischemic injury, Chinese Pharmacology and Clinical Medicine 2007; 23(2): 19-21; Xu Wei, Liu Chunqing, Ma Tao et al. Protective effects of ginsenoside Re on dopaminergic neurons in MPTP-induced Parkinson's disease model mice, Journal of Shenyang Pharmaceutical University, 2005, 22

(1): 36-44; ginsenoside composition and preparation method, invention patent, application number 200610046083.3; compound 20S-ginsenoside R 2 in preparation of antidepressant medicine, invention patent, application number 200810043537.0).

Rhizoma Coptidis, with diarrhea, dampness, detoxification, insecticidal effects, modern pharmacological studies have shown that In addition to strong antibacterial activity, it also has a variety of pharmacological effects such as anti-inflammatory, hypolipidemic, hypoglycemic, anti-tumor, anti-ulcer, and nervous system protection. It has been reported that Coptis has protective effects in cerebral ischemia and reperfusion injury through anti-inflammatory effects (Ki-Yeon Yool, In Koo Hwang2, Jong Dai Kim3, etc., Antiinflammatory Effect of the Ethanol Extract of Berberis koreana in a Gerbil Model of Cerebral Ischemia /Reperfusion Phytother. Res. 2008, 22, 1527-1532 ).

 The scorpion is the fruit of the Gardenia j匪 inoides Ellis. It is bitter and cold and non-toxic. Into the heart, liver, lungs, stomach. It has the effects of purging fire, removing heat, diuresis, cooling blood and detoxification. Indications of fever, irritability, jaundice, gonorrhea, diabetes, red eyes, sore throat, vomiting blood, blood stasis, blood stasis, hematuria, heat sores, sprains and swelling. Clinically used for acute jaundice hepatitis, hemostasis, contusion and other diseases. Modern pharmacological studies have shown that in addition to traditional pharmacological effects such as antipyretic, antibacterial, sedative, analgesic, choleretic, anti-inflammatory, hemostasis, liver protection, and soft tissue damage repair, medlar has anti-oxidation, anti-asthma, and hypolipidemic effects. , antihypertensive, anti-allergic, anti-atherosclerosis, anti-viral, anti-depressive anxiety, nervous system protection, etc. (Study on the chemical constituents and pharmacological effects of gardenia, North Pharmaceutical, 2008, 5 (3): 44-45, 51; Huang Shisun, Wu Yuyue, Overview of Modern Pharmacological Research and Clinical Application of Xunzi, Internal Medicine, 2010, 5 (5): 534-536; Liu Guomin, Guo Suhua, Cheng Weiming, New Progress in the Pharmacological Action and Mechanism of Gardenia, Strait Pharmaceuticals, 2008, 20 (1 1 ): 8-10; Use of Gardenia Extract in the Preparation of Drugs for Prevention and/or Treatment of Depression Worries, Chinese Invention Patent, Application No. 200810016679.8).

 The classic compound Huanglian Jiedu Decoction shared by Huanglian and Scorpion is originally contained in the "Outside Taiwan Secrets". It is composed of berberine, Phellodendron, Astragalus and Scorpion. It is regarded as the representative of detoxification and detoxification. It has the effect of purging fire and detoxification. Practical heat and poison, Sanjiao hot Sheng card. It has been reported that Huanglian Jiedu Decoction can reduce brain damage and inflammatory infiltration in cerebral ischemia and reperfusion (Young Sun Hwang a, Chan Young Shin b, Youngbuhm Huh et al., Hwangryun-Hae-Dok-tang (Huanglian). -Jie-Du-Tang) extract and its constituents reduce ischemia-reperfusion brain injury and neutrophil in filtration in rats. Life Sciences 2002, 71 : 2105-21 17).

 However, the inventors have noted that there is still a lack of pharmaceutical preparations that can be used for the treatment of brain and nerve damage diseases. Summary of the invention

The composition of the present invention is a key path machine for "poisoning the brain and collaterals", and is proposed by treating syndromes and correcting the evils of the Chinese medicine. Yiqi Jiedu Recipe is a traditional Chinese medicine compound preparation that has not been reported before. It has not been reported for the prevention and treatment of brain and nerve damage diseases in Chinese medicine. Yiqi Jiedu Recipe consists of ginseng, berberine and medlar. Among them, ginseng is a medicinal herb, it is a great qi, and it can replenish the five viscera. It is very suitable for the treatment of qi deficiency; berberine is a drug, it is a good product of diarrhea and detoxification, compatible with ginseng, one supplement and one diarrhea, The scorpion is the adjuvant medicine. The medicine is good for clearing the heart and removing troubles, cooling blood and detoxifying, assisting the diarrhea and detoxification of berberine, and at the same time, its sexual flexion descends, and drives the evil spirits out of the urine, so that the evil of heat and poison Way out. This three-flavor compatibility can effectively detoxify evil on the basis of righting up, and plays a very good synergistic therapeutic effect. Excessive release of excitatory amino acids and neurotransmitter abnormalities, oxidative stress, free radicals, inflammatory cascades, mitochondrial dysfunction, etc. are common pathological mechanisms of brain injury, ginseng grows in qi, neuroprotection, berberine and scorpion are longer than detoxification , the three compatibility can be rationally compatible and mutually complementary.

 Accordingly, the technical problem to be solved by the present invention is to provide a composition of a traditional Chinese medicine compound or extract capable of preventing or treating brain and nerve damage diseases.

 The invention relates to a composition of a traditional Chinese medicine compound or an extract, which is composed of ginseng, berberine and medlar.

 The composition according to the present invention as described above, wherein the content of ginseng, berberine, and wolfberry is calculated as 100 parts by weight: ginseng, 1-99 parts, berberine, 1-99 parts, wolfberry, 1-99 parts Preferred ginseng, 2-75 parts, berberine, 2-75 parts, hazelnut, 2-75 parts; more preferably ginseng 3-50 parts, berberine, 3-50 parts, wolfberry, 3-50 parts.

 The composition according to the present invention as described above, wherein the content of the ginseng extract, the coptis extract, the hazelnut extract is 100 parts by weight based on the total weight: ginseng extract, 1-99 parts, berberine extract, 1- 99 parts, hazelnut extract, 1-99 parts; preferred ginseng extract, 2-75 parts, Coptidis Rhizoma extract, 2-75 parts, hazelnut extract, 2-75 parts; more preferably ginseng extract 3-50 Serving, Coptidis Rhizoma Extract, 3-50 parts, Gardenia Extract, 3-50 parts.

 The composition according to the present invention as described above, wherein the weight ratio of ginseng, berberine and medlar is 3:2:2 to 0.5, preferably 3:2:1, more preferably 3:2:2.

 The composition according to the present invention as described above, wherein the weight ratio of the ginseng extract, the coptis extract, the hazelnut extract is 3:2:2-0.5, preferably 3:2:1, more preferably 3:2:2 .

 The composition according to the present invention as described above, comprising a traditional Chinese medicine compound or extract as an active ingredient and any one or more pharmaceutically acceptable carriers such as an antioxidant, a complexing agent, a filler, a matrix material, etc. .

 The composition according to the present invention as described above, which can be formulated into a pharmaceutically acceptable dosage form, such as a tablet, a granule, a capsule, an injection such as a powder, according to a specific need and a pharmaceutically acceptable carrier. Freeze dry powder, infusion, etc.

 The invention further relates to the composition as described above for the preparation or treatment of brain and nerve damage, such as cerebral vascular dementia or ischemic cerebrovascular disease (eg transient local ischemic attack, cerebral infarction or stroke), neurodegenerative Use in diseases of the brain, such as Alzheimer's disease or Parkinson's disease, bacterial or viral encephalitis, or depression. DRAWINGS

Figure 1: TTC staining results of cerebral ischemic area of cerebral ischemia rats after 24 hours of MCAO surgery, A: sham operation group; B: model group; C: positive control group (ginkgo biloba extract Egb761 tablets) Group) (4mg/kg); DF: 1, 2, 3 (10g crude drug / kg). Figure 2: Effect of the composition of the invention on the volume of cerebral infarction in rats at 12 h and 24 h after MCAO surgery (n = 10).

detailed description

 The invention will be further illustrated by the following examples in which the advantages and features of the invention will become more apparent. However, these examples are merely exemplary and do not constitute any limitation on the scope of the invention. It will be understood by those skilled in the art that the details and forms of the present invention may be modified or substituted without departing from the spirit and scope of the invention, and such modifications and substitutions fall within the scope of the invention.

 Preparation process of traditional Chinese medicine compound and its preparation

 Preparation of traditional Chinese medicine compound

 The ginseng, berberine and medlar pieces with a weight ratio of 3:2:2-0.5 were pulverized, and the powder of huanglian decoction pieces was extracted with 50% ethanol under reflux for 3 h/time. The ethanol extract was concentrated and concentrated with concentrated hydrochloric acid. pH=2, 10%-12% NaCl solid was added, fully stirred to completely dissolve, left overnight, suction filtered, and the precipitate was washed with a small amount of distilled water and dried at 50 to 60 ° C to obtain a Coptidis extract. The ginseng and medlar pieces are further boiled four times with water for 2 hours, and the amount of water added is 12 times of the amount of the medicinal material; the decoction is combined, filtered, and concentrated to a relative density of 1.04-1.05 (50 ° C). Clear the paste, add ethanol to make the alcohol content up to 70%, refrigerate (5~10 °C) overnight, filter, the filtrate is recovered ethanol, and concentrated to a relative density of 1.32~1.35 (50 °C) of the clear paste, minus Press dry (70 ° C), the dry paste is pulverized into fine powder, and an extract mixture of ginseng and wolfberry is prepared, mixed with the extract of Coptidis Rhizoma, and uniformly stirred to obtain a homogeneous mixture.

 Preparation of traditional Chinese medicine compound preparation

 Preparation of traditional Chinese medicine compound tablets

 Take the above homogeneous mixture, add appropriate amount of medicinal adjuvant starch, magnesium stearate, etc., mix well, and then tablet to prepare a tablet containing the above traditional Chinese medicine compound.

 Preparation of Chinese Medicine Compound Capsules

 The above homogeneous mixture is taken, and the amount of the medicinal adjuvant starch is added, and the mixture is thoroughly mixed, and then filled into capsules to prepare a capsule containing the above-mentioned traditional Chinese medicine compound.

 Preparation of traditional Chinese medicine compound granules

 A plurality of the above homogeneous mixtures are taken, and an appropriate amount of the medicinal adjuvant starch is added to prepare a granule containing the above-mentioned traditional Chinese medicine compound.

 Preparation of traditional Chinese medicine compound injection

 The above homogeneous mixture was taken, and an appropriate amount of pure water for injection and polysorbate 80 was added to prepare an injection containing the above-mentioned traditional Chinese medicine compound.

 Preparation of powder injection for traditional Chinese medicine compound injection

The above homogeneous mixture is taken, and an appropriate amount of the water-soluble medicinal auxiliary material for injection is added to prepare a powder injection for injection containing the above-mentioned traditional Chinese medicine compound. Preparation method of traditional Chinese medicine extract and preparation thereof

 Preparation of Chinese herbal extracts

 According to the preparation method of Patent No. 00110418.7, the ginseng total saponin extract is obtained, and according to the preparation method of Patent No. 201010156823.5, the scorpion glycoside extract having 99% saponin content is obtained, according to the preparation method of Patent No. 200710020332.6, The berberine hydrochloride having a content of 98% is uniformly mixed according to the weight ratio of ginseng total saponin, berberine hydrochloride and geniposide of 3:2:2-0.5 to obtain a mixture for use.

 Preparation of Chinese herbal extract tablets

 The above mixture is added, and an appropriate amount of the medicinal adjuvant starch, magnesium stearate or the like is added, and after thorough mixing, the tablet is tableted to prepare a tablet containing the above-mentioned traditional Chinese medicine extract.

 Preparation of Chinese Herbal Extract Capsules

 The above mixture is taken, and an appropriate amount of the medicinal adjuvant starch is added, and the mixture is thoroughly mixed, and then filled into capsules to prepare a capsule containing the above-mentioned traditional Chinese medicine extract.

 Preparation of Chinese medicine extract granules

 A plurality of the above mixture is taken, and an appropriate amount of the medicinal adjuvant starch is added to prepare a granule containing the above-mentioned traditional Chinese medicine extract. Preparation of traditional Chinese medicine extract injection

 The mixture was mixed, and purified water and polysorbate 80 were added in an appropriate amount to prepare an injection containing the above-mentioned traditional Chinese medicine extract.

 Preparation of powder injection for injection of traditional Chinese medicine

 The above mixture is added, and an appropriate amount of the water-soluble medicinal adjuvant for injection is added to prepare a powder injection for injection containing the above-mentioned traditional Chinese medicine extract. Example 1 Experimental study on the protective effect of the composition of the present invention on acute focal cerebral ischemia

 Establishment of an animal model of acute focal cerebral ischemia

Male Sprague-Dawley rats weighing 250-270 g were used according to Zea Longa et al. (Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20: 84-91). In the middle cerebral artery of rats, the method of blocking the plug line, 10% chloral hydrate solution 400mg/kg, anesthetized animals were injected intraperitoneally. The rat was fixed in the supine position, the skin was cut in the middle of the neck, and the layers of tissue were bluntly separated to expose the right common carotid artery (CCA). Separate to the internal carotid artery (ICA), external carotid artery (ECA) after a bifurcation, carefully separate to avoid damage to the vagus nerve and trachea, set the line for use. The ipsilateral external carotid artery was isolated and ligated at approximately 0.8 cm from the ECA. An arterial clip was placed at the proximal end of the CCA. A "V"-shaped incision with a diameter of about 2 mm was made between the ECA ligation and the bifurcation. The nylon thread was gently inserted into the CCA from the incision, and the artery clip was released. The inner and outer carotid bifurcations enter the internal carotid artery, and the nylon thread is slowly pushed into the cranial direction of the ICA. The insertion depth is about 18.5±0.5 mm to the micro-inductance, so that the nylon thread end passes through the MCA start point. The finer anterior cerebral artery, at this point, the blood flow obstruction of the left middle cerebral artery is achieved, and the ICA is ligated to fix the nylon thread and prevent bleeding. Stitched, the nylon thread stump is 1 cm longer than the skin.

 The sham operation group only performed preoperative anesthesia and vascular separation, without ligation and introduction of a wire plug. The room temperature was maintained at 24-25 °C during the operation, and the biological function test system was used for animal breathing and ECG monitoring.

 Animal grouping and administration

 SD rats were randomly divided into six groups: sham operation group, model group, Ginkgo biloba extract tablets positive control group (Ginado Ginkgo biloba extract tablets, Egb761, Dr. Weimar Shupei, Germany), the combination of the present invention Drug administration group 1, 2, 3. Positive control group administration method: Take 1 piece of Ginkgo biloba extract tablets (40mg) dissolved in 40ml normal saline, prepare lmg/mL, and give SD rats 1mL by gavage, the dosage is 4 mg/kg. Administration method of the drug-administered group: ginseng, berberine, and wolfberry pieces were administered by gavage according to the weight ratio of 3:2:0.1 (administering group 1), 3:2:2 (administering group 2), 3:2:0.5 (Determination group 3) The extract obtained by the above-mentioned traditional Chinese medicine compound preparation process was administered in an amount of 10 g of crude drug/kg. The drug-administered group and the positive control group were intragastrically administered 10 minutes before the anesthesia, and the animals were sacrificed at 4, 12, and 24 hours after surgery, and the following indicators were tested. Neurobehavioral Bederson's functional evaluation

Neurobehavioral observations were performed before sacrifice, according to the method of Bederson et al. (; Bederson JB, Pitts LH, Tsuji M, et al., Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke 1986; 17:472-476), take the rat tail off the ground about 1 foot, observe the condition of the two forelimbs; place the rats on the level ground, push the shoulders, observe the difference of the resistance on both sides; the rats are placed on the ground, observe the Walking situation. Using a five-point scale (0-4 points), the higher the score, the more serious the neurobehavioral damage. The statistical results of Bederson's functional scores are shown in Table 1. The data of each group were compared using the rank sum test, and Ρ<0.05 was considered statistically significant.

Table IBederson's functional score statistics (^±sd, n=10) positive

 Sham surgery model

 Item Level Control Drug Delivery Group 1 Drug Delivery Group 2 Drug Delivery Group 3 Group

 Group

 0 6

 1 1 4

 2 1 3 2

 12h-l 3 5 2

 4

 Rank

 P value # #

 0 7 4 4 5

1 2 0 4 2 1

2 1 2 2

 12h-2 3 4 1 1 1

 4

 Rank

 P value ## # ## ##

0 10 0 5 5 1 0

1 0 4 4 3 6 7

2 0 0 0 1 3 2

24h-l 3 0 1 0 0 0 0

 4 0 3 0 0 0 0

Rank

 P value ## ## ## #

0 8 0 3 5 5 3

1 2 4 3 2 2

2 4 0 0 0 1

24h-2 3 0 0 0 0 0

 4 2 0 0 0 0

Rank

 P value # ## ## ##心

 Product P value ## ## ## 分

Note: Non-parametric rank sum test, test, compared with the control group: ^# *P<0.01; compared with the model group: #P<0.05,

##P<0.01

Conclusion: It can be seen from Table 1 that the model group is statistically different from the sham operation group, indicating that the modeling is successful (Ρ<0.01). The positive control group (4 mg/kg) and the drug-administered group 1, 2, 3 significantly improved the neurological symptoms at 12h and 24h after focal cerebral ischemia in animals (P<0.05, P<0.01). Evaluation of the improvement of infarct volume

After functional scoring, the rats were decapitated and the brain tissue was quickly placed in a refrigerator at -20 °C. After lOmin, the room temperature was set. The olfactory bulb, cerebellum and low brain stem were removed and cut into 2 mm crowns according to the brain localization map. Six brains were continuously coronal sections. Then quickly place the brain slices in 5ml containing 2% TTC (triphenyltetrazolium chloride) and

In a 0.1 ml solution, incubate at 37 ° C for 30 min in the dark, and flip the brain slices once every 5 min. After TTC staining, the normal tissue was rose red, and the infarcted tissue was unstained and white. Each group of brain slices was arranged neatly, photographed and saved, and processed by image analysis system software and counted. The infarct area of each brain slice was calculated, multiplied by the thickness of each brain slice by 2 mm, and the infarct area of each brain slice of each animal was multiplied by The thickness is added, which is the volume of cerebral infarction.

 Conclusion: As shown in Figures 1 and 2, after cerebral ischemic injury in rats, no cerebral infarction occurred in the sham operation group, and the infarct area appeared in the model group and the TTC stained grayish white. The administration group 1, 2, 3 can significantly reduce the gray area of the cerebral infarction, reduce the volume of cerebral infarction, and significantly improve the cerebral infarction.

 From the above experiments, it is known that the composition of the present invention has a good protective effect against acute cerebral ischemic injury. Example 2 Experimental study on the protective effect of the composition of the present invention on senile dementia (AD) animals

 Incubation of β-amyloid (Αβ^ο)

 Dilute Αβ^ο into 2ug/ul with sterile saline and incubate at 37 °C for 1 week to make it condensed.

Αβι

 Establishment of rat dementia model induced by Αβ^ο

 250-270 g of healthy male AD rats were anesthetized and fixed on a rat stereotaxic instrument. The skin was routinely sterilized, the skin was cut, the anterior sputum was exposed, and a small syringe was used to the left hippocampus of the rat (with a Bregma point of 0). , AP-3.0mm, ML2.0mm, DV-2.9mm) Slowly inject the condensed state Αβ^ 5μ1, leave the needle for 5min to make the Αβ fully diffuse, then slowly withdraw the needle and suture the incision. The sham operation group only performed preoperative anesthesia, cut the skin, and sputum before exposure, without injection Αβ^^

 Experimental grouping and administration

Thirty-two healthy male SD rats of 250-270 g were randomly divided into a sham operation group, a Αβ model group, a ginkgo biloba extract tablet positive control group, a composition administration group of the present invention 1, 2, 3, and 5 rats in each group. Positive control group administration method: Take 1 tablet (40 mg) of Ginkgo biloba extract tablets, dissolve it in 4 ml of normal saline, prepare 1 mg/mL, and give 1 mL of SD rats by gavage, and the dosage is 4 mg/kg. The administration method of the administration group: the extract obtained by the preparation method of the traditional Chinese medicine compound according to the weight ratio of 3:2:0.5 by the gavage, the ginseng, the berberine and the scorpion decoction pieces respectively, the dosage of the medicine is 20 g of the crude drug/kg (the administration group 1) ), 10 g of crude drug/kg (administered group 2) and 5 g of crude drug/kg (administered group 3). After the model was completed, the Αβ model group and the sham operation group were injected with the same volume of normal saline, and the other groups were given the corresponding drugs by gavage. Morris Water Maze Test

 Water maze test was started 30 days after surgery. The water maze is a pool with a diameter of 90cm and a height of 40cm. The water depth is 30cm. Four water inlet points are marked on the wall of the pool. The water is divided into four quadrants. A plexiglass circular platform is placed in the center of one of the quadrants. The platform is 2cm below the water surface. The water temperature (24 ± 2) ° C, covered with opaque thick and even white plastic foam, the reference material around the pool remains unchanged. The tests include:

 1) Positioning navigation test: It is used to measure the ability of rats to acquire and learn from water fans. For 5 days, two time periods in the afternoon and the afternoon, four times in each time period, the rats were placed into the water from four different marking points, and the time required to find the platform within 2 minutes (evacuation latency) was recorded. If the platform fails to find the platform within 2 minutes after entering the water, place it on the platform for 10 s and escape the incubation period for 2 min. Each training interval is 60s.

 2) Space exploration test: It is used to measure the ability of the rats to maintain the memory of the platform space after learning to find the platform. After the end of the navigation test, the platform was removed on the sixth day. At any water inlet point, the rat was placed in the water facing the pool wall, and the number of times across the original platform in 2 minutes was recorded.

 One-way analysis of variance (COne-Way ANOVA) was used, and P < 0.05 was considered statistically significant. The results of the Morris water maze test are shown in Table 2.

 Table 2 Average escape latency in positioning navigation test (s, x ± sd, n=5) Group 1 day 2nd day 3rd day 4th day 5 days sham operation group 74.89±21.30 38.10±14.80 22.60±9.20 21.02± 8.73 17.80±7.71

Αβ model group 77.80±22.40 59.88±15.50 47.2±11.80** 48.11±11.52 46.15±15.00** Positive control group

75.30±22.31 40.10±15.20 23.01±9.30 ^## 21.04±8.75 ^## 17.90±7.80. Administration group 1 75.10±22.10 39.40±15.0 22.8±9.32 Pu 21.30±8.80 ^## 17.90±7.80 Administration group 2 75.20±23.00 39.43 ±15.30 23.10±9.35 21.50±8.90 ^## 17.95±7.85 ^## administration group 3 76.30±23.50 39.90±15.50 23.70±9.45 22.10±9.03 18.30±8.05

0.01, ^ffff P < 0.01

 Table 3 Results of space exploration experiments in Morris water maze rats ± ^ n=5 ) Group latency (S ) Number of positions through the original platform Sham operation group 10.26±4.37 3.59±1.37

 Model group 29.16±12.64 1.42±1.04

Positive control group 16.03±9.19 2.75±1.16 ^#

Drug administration group 1. 13.74±8.66 2.94±1.42 ^#

 Drug administration group 2 16.76±9.24 2.81±0.98*

Administration group 3 18.69±10.27* 2.79±1.03 ^#

Note: ^# Ρ < 0.05, Ρ < 0.01 As can be seen from Table 2, the rats did not change significantly in the first 2 days after administration of the composition of the present invention. From the third day, the model group was compared with the sham operation group: "Ρ < 0.01, indicating successful modeling; the administration group and Αβ The model group comparison: ## Ρ < 0.01, which indicates that the administration groups 1, 2, and 3 can significantly shorten the average escape latency from the last three days. As can be seen from Table 3, the administration group can significantly shorten the spatial exploration latency of rats. across the internet to increase the number of the original position, the compositions of the present invention _{1 4} _ ₍₎ induced rat model of AD has protective effects alumina and dementia model mice induced by administration method

 The experimental animals were selected from 22-month-old Kunming mice, male and female, and randomly divided into 6 groups: normal control group, aluminum trichloride model group, aluminum trichloride plus ginkgo leaf extract tablets positive control group, three Aluminum chloride plus the compositions of the invention were administered to groups 1, 2, and 3. Positive control group administration method: Take 1 piece of Ginkgo biloba extract tablets (40mg) dissolved in 4ml normal saline, prepare 1mg/mL, and give mice lmL by intragastric administration, the dosage is 4 mg/kg. The administration method of the administration group: the extract obtained by the preparation method of the traditional Chinese medicine compound according to the weight ratio of 3:2:0.5 by the gavage, the ginseng, the berberine and the scorpion decoction pieces respectively, the dosage of the medicine is 20 g of the crude drug/kg (the administration group 1) ), 10 g of crude drug/kg (administered group 2) and 5 g of crude drug/kg (administered group 3). The mice in the aluminum trichloride model group were intragastrically administered with a 10 mg/ml aqueous solution of aluminum chloride (manufactured by Shanghai Jinshan Chemical Plant, batch number 100093) at 200 mg/kg body weight/day for 90 days. Ginkgo biloba extract tablets and the present invention compositions were administered to the positive control group and the administration group while the dementia model was established, and administered intragastrically for 90 days. The normal control group was given the same dose of physiological saline.

 Platform test

TT-2 mouse platform automatic control instrument (developed by the Institute of Materia Medica, Chinese Academy of Medical Sciences). The jumping table reflection box is a 90x 18x60cm ³ plexiglass box, which is separated into five by a transparent plate. The bottom of the box is a copper grid, and the voltage is 36V. Each built-in rubber pad with a height of 3.5cm and a diameter of 3.5cm is used as a mouse to avoid electric shock. Safe area. Before the experiment, 3 angels were familiar with the laboratory environment, and the room was kept quiet at a temperature of 25 °C. During the experiment, the mice were placed in a reflective box for 5 min, and then the current was passed at 0.25 mA. The mice were placed on a platform. The mice had the habit of jumping from a height. If they jumped, they were shocked and generated memories. . When the platform has a latency of more than 180 s during training, the number of times the mouse jumped off the platform within 5 minutes (the number of errors) was recorded, and the mice were divided into 6 groups based on the number of errors (normal control group, model group, positive control). Ginkgo biloba extract tablets group, administration group 1, 2, 3, positive control and administration groups 1, 2, 3), 10 mice in each group, the intelligence level of the 6 groups of mice was close. During the experiment, the number of times the mice jumped from the stage (the number of errors) within 5 minutes was carefully determined in detail, which was used as an observation index for learning and memory consolidation in mice.

One-Way ANOVA analysis, Ρ<0.05 was considered statistically significant. The results of the platform test are shown in Table 4. Table 4 Comparison of learning and memory ability changes in mouse platform test (±S, n=10)

Group memory acquisition error number of memory memory consolidation error number of normal control group 1.26 ± 0.73 1.80 ± 0.40 model group 6.78 ± 2.40 * 5.90 ± 1.08 ^# positive control group 3.60 ± 2.00 4.78 ± 0.63 ^# drug group 1 2.87 ± 1.90 ^## 3.50±0.88 administration group 2 3.48±2.05 4.10±0.95 administration group 3 4.00±2.10 4.88±1.02 ^#

0.05, ^ff P < 0.05, ^ffff P < 0.01

As can be seen from Table 4, the model group was compared with the drug-administered group: ^# Ρ < 0.05, indicating successful modeling; the drug-administered group was compared with the model group: ^# P < 0.05, P < 0.01, indicating that the composition of the present invention can significantly reduce the mice The number of errors in the dementia model has a significant improvement in the learning and memory ability of mice. Example 3 Experimental study on the protective effect of the composition of the present invention on Parkinson's animal model

 Parkinson animal model establishment and administration method

 A mouse model of Parkinson's disease was established by intraperitoneal injection of pesticide paraquat (PQ). A total of 120 male Kunming mice, 7 weeks old, were randomly divided into 6 groups, 20 in each group. The blank control group: 10 mL/kg body weight of distilled water once a day, once a week, 10 ml/kg body weight of normal saline was injected once a week; the other 5 groups were intraperitoneally injected with PQ 10 mg/kg once a week, and colchicine was administered once a day. It was administered at a dose of 4 times that of an adult, that is, 0.2 mg/kg, for 7 weeks. After successful modeling, the positive control group: Metoprolol was administered once a day, according to the dose of 7.5 times of the adult dose, that is, the dose was 125 mg/kg, and the administration was continued until one week after the modeling; 2, 3: The mice of the present invention were intragastrically administered with 20 g of crude drug/kg, 10 g of crude drug/kg, and 5 g of crude drug/kg (administering method is the same as in Example 2), and continued to be administered for one week after modeling. Serve under the same conditions, observe the behavioral changes of mice, and then test the content of monoamine neurotransmitters in the substantia nigra of mice, dopamine (DA), norepinephrine (NE), serotonin (5 -HT) detection.

 Rotating rod experiment

 Normal mice have the ability to climb, which requires proper grip and motor coordination. We used a rotating rod test to detect motor dysfunction in mice. The mice were placed on a rotating rod tester (YLS-2C, Beijing Ji'an Deer Technology Co., Ltd.), and each time, 6 rats were rotated at a rate of 20 revolutions per minute, and the number of times the mice fell within 3 minutes was recorded.

 Self-determination

 The mice were placed in the activity box of the autonomous activity tester, and 6 mice were simultaneously measured at a time, 1 mouse in each box, and the mice were allowed to acclimate for 3 min before starting the test, when the computer entered the measurement procedure. After that, the number of activities and standing of the mouse within 10 minutes was automatically recorded by the computer.

One-Way ANOVA analysis, Ρ<0.05 was considered statistically significant. The behavioral test results are shown in Table 5. Table 5 Behavioral test results (;^ ±S, n=20)

 Rotating stick within 3min number of horizontal activities number of standing activities total number of activities

 Drop number (times) (times) (times)

 (times) blank control group 2.4±0.6 8.4±0.8 76.3±14.5 88.6±15.8

PQ model group 6.8±0.9** 3.5±0.6** 26.8±12.1** 29.6±12.3** Positive control group 3.4±0.7 7.5±0.8 ^## 56.4±10.2 66.4±13.6 ^## administration group 1 3.5±0.4 ^{# #} 6.8±0.7 58.7±19.6 64.8±17.2 ^## administration group 2 4.8±0.8# 5.5±0.9 ^# 63.6±14.3 54.2±11.8 Administration group 3 5.5±0.7 ^# 4.0±0.8 65.5±10.3 ^# 40.3±15.4 ^#

0.01, ^ff P<0.05, P<0.01

As can be seen from Table 5, the model group was compared with the blank control group "P < 0.01, indicating successful modeling; after the administration of the composition of the present invention, the number of times the mouse dropped from the rotating rod was reduced, and the number of activities was significantly increased. Group comparison: ^# P < 0.05, ^## P < 0.01, indicating that the composition of the present invention can significantly improve the behavioral ability of Parkinson's mice.

 The results of the detection of monoamine neurotransmitters are shown in Table 6.

 Table 6 Results of determination of DA, NE, and 5-HT content (±s, n=10) Group 5-HT(ug/g) DA(ug/g) NE(ug/g) Blank control group 19.74±10.6 27.96± 10.12 23.79±8.6

PQ model group 53.4±7.8** 64.38±9.64* 44.86±10.5** Positive control group 36.53±8.4 ^## 42.28±10.4 ^## 33.24±7.8 Administration group 1 35.89±9.5 40.76±8.8 ^## 30.67±8.4 Administration group ^{2 42.56 ± 6.9 # 49.25 ± 12.3} 33.12 ± 9.3 administration group ^{3 46.88 ± 7.2 50.62 ± 11.2 #} 39.88 ± 5.4 Note: P <0.05, P <0.01 , # P <0.05, P <0.01

 As can be seen from Tables 5 and 6, the composition of the present invention can significantly reduce dyskinesia in PQ-induced Parkinson's mice, reduce excessive release of monoamine neurotransmitters, and improve Parkinson's symptoms. Example 4 Experimental study on the protective effect of the composition of the present invention on an animal model of vascular dementia

 Animal model establishment

A rat model of bilateral vascular avascular dementia was permanently ligated. After SD rats were intraperitoneally injected with 3.5% chloral hydrate 10ml/kg, the supine position was fixed, the midline incision was made in the neck, and the bilateral common carotid arteries were separated to avoid The cervical sympathetic nerve and the vagus nerve were damaged, and they were permanently ligated with the line 1, and the wound was sutured and the cage was incubated. The sham operation group was to separate the bilateral common carotid arteries, but not to be ligated.

 Animal administration method

 One month after the operation, the rats were divided into six groups: the sham operation group, the model group, the ginkgo biloba extract tablets positive control group (4 mg/kg), and the Chinese medicine extract administration group of the present invention 1, 2. 3 (20 g crude drug/kg, 10 g crude drug/kg and 5 g crude drug/kg) (the administration method is the same as in Example 2).

 Morris water maze learning memory test

 The water maze is a pool with a diameter of 90cm and a height of 40cm. The water depth is 30cm. Four water inlet points are marked on the wall of the pool. The water is divided into four quadrants. In one of the quadrants, a plexiglass circular platform is placed, and the platform is 2cm below the water surface. The water temperature (24 ± 2) ° C, covered with opaque thick and even white plastic foam, the reference material around the pool remains unchanged. The tests include:

 1) Positioning heading experiment: It is used to measure the ability of rats to acquire water memory and learn from memory. Place the platform in the middle of a certain quadrant of the labyrinth, select one of the other three quadrants, and experiment twice a day, each time using a different water inlet point, each training for 1 minute. If the animal has not found the platform within 2 minutes, take the animal to the platform and let it stand for 15 seconds. If the rat finds the platform within 2 minutes, let it stand on the platform for 15 seconds. training. After five days of training, the escape latency and swimming path of each animal were recorded separately.

 2) Space search experiment: It is used to measure the ability to maintain the memory of the platform space position after the rats learn to find the platform. After the end of the navigation test, the platform was removed on the sixth day. Each rat was only swam for 2 minutes. The time of the first passage of each rat, ie the incubation period, was recorded with a stopwatch, and each rat was recorded. The number of passes through the platform within 2 minutes, if the incubation period of a rat is shorter and the number of positions passing through the original platform is more, the spatial positioning ability and learning and memory function of the rat are good.

 Morris water maze experiments were started 14 days after administration. First, the heading experiment was carried out. Each rat was trained twice a day for a training period of 120 s and continuous training for 5 days. The parameters were recorded. A spatial search experiment was then performed and the platform was removed on the sixth day, recording the latency of the rat and the number of passes through the platform within 2 minutes.

 One-way analysis of variance (One-Way ANOVA) analysis, Ρ < 0.05 was considered statistically significant. The results of the water maze experiment are shown in Tables 7, 8, 9, and 10.

 Table 7 Vascular Dementia Rats Positioning Heading Experiment Escape Latency (±s) Number of Animals Escape Latency (s)

 Day 3 Day 4 Day 5 Sham-operated group 12 13.38±5.88 12.04±10.26 9.23±5.32 Model 11 60.24±20.12 ##

Group 42.32±19.56 ^ffff 34.42±17.86 positive control

4 mg/kg 11 20.32±14.24 17.63±10.25 14.68±11.54 Group Administration group 1 20 g raw

 12 18.76±12.45 15.56±8.24 13.96±8.71 medicine /kg

 Administration group 2 10 g raw

 11 24.22±15.68 21.25±13.78 21.21±11.37 medicine /kg

 Administration group 3 5 g crude drug

 11 35.79±23.12 26.38±12.39 24.42±9.76 /kg

 Note: P<0.05, P<0.01, P<0.01

As apparent from Table 7, comparing the model group with the sham ^{group, # P <0.05, P <} 0.01, described successful modeling, comparing administration group and model group, * P <0.05, ** P <0.01, combination of the present invention described The substance can significantly reduce the escape latency of vascular dementia rats.

 Table 8 Experimental navigation of vascular dementia rats (; i±s)

 Animal tour (cm)

 Group number

 (only) third day fourth day fifth day sham surgery

 12 385.64±214.32 298.79±203.56 212.54±132.74 Group

Model group 11 1647.76±688.43 1018.04±463.74 ^## 921.64±366.54 ^##正对

4 mg/kg 11 543.22±216.87" 356.28±144.84 ^# 346.43±235.2

Administration group 20 g crude drug

 12 606.42±341.64** 394.29±243.66* 332.96±164.82**

1 / kg

Administration group 10 g crude drug

 11 787.21±463.50* 492.55±276.72* 406.78±203.54*

2 /kg

Administration group 5 g crude drug

 11 899.54±532.04* 754.74±448.32 592.16±243.12* 3 /kg

 Note: P<0.05, P<0.01, P<0.01

 As shown in Table 8, the model group compared with the sham operation group, #P<0.05, P<0.01, indicating successful modeling, the drug-administered group compared with the model group, *P<0.05, **P<0.01, indicating the combination of the present invention The material can significantly reduce the run of the vascular dementia rats in the water maze experiment.

 Table 9 Results of space exploration experiments in rats with vascular dementia (±s)

Number of animals passing through the original platform position subgroup

Incubation period (s) sham operation group 12 11.37±5.29 3.68±1.65 model group 12 28.06±13.57 1.35±1.23 Positive control ^ _n

 Group 4 mg/kg 12 15.76±8.69 2.88±1.36 Administration group 1 20 g crude drug

 12 14.98±9.01 3.00±1.24 /kg

 Drug administration group 2 10 g crude drug

 12 17.04±7.32 2.76±1.38" /kg

 Administration group 3 5 g crude drug /kg 12 17.58±10.62 2.65±1.44 Note: *P < 0.05, **P < 0.01

 As can be seen from Table 9, the administration group was compared with the model group, *P<0.05, **P<0.01, indicating that the incubation period of the space exploration experiment after the administration of the composition of the present invention was significantly shorter than that of the model group, and the number of the original platform was significantly shorter. increase.

 Table 10 Effects of different Chinese medicines on the incubation period of the VD rats in the search for platform (±s, n=8)

Group search platform latency (S)

 1 day 2nd day 3rd day 4th day 5 days normal group 35.90±24.36 25.90±25.76 23.54±20.12 15.9±9.86 10.8±6.87 sham operation group 38.60±22.78 28.60±31.42 26.38±20.56 17.6±11.35 15.3±10.34 Model group 67.03±32.74* 62.6±39.88** 56.5±27.58** 42.6±23.08** 38.6±24.64** Administration group 1 37.36±28.32 29.46±21.26 27.14±23.29 20.7±16.72 18.2±10.72

0.05, P < 0.01, ^ffff P < 0.01

As can be seen from Table 10, the drug-administered group was compared with the model group, ^## P < 0.01, indicating that the composition of the present invention can significantly shorten the latency of the rat seeking platform.

 From the above experimental results, it can be seen that the administration of the composition of the present invention can significantly improve the learning and memory ability of the vascular dementia rats.

 Dark test

 The rats were placed in a bright room and allowed to move freely for 3 min in the box. Those who do not enter the dark room within 3 minutes are excluded (not energized during pre-selection). After 1 h, the training was officially conducted. The time required for the rats to enter the dark room to receive electric shock was recorded as the incubation period, and the number of electric shocks (the number of errors) within 5 minutes was recorded as the academic score. After 24 hours, the above process was repeated, and the latency and the number of errors were recorded. Memory scores. The voltage is 36 V and the latency indicator is in seconds (S).

One-Way ANOVA analysis, Ρ<0.05 was considered statistically significant. The results of the darkness test are shown in Table 11. Table 11 Results of darkness test in rats with vascular dementia (±S, n=10) al learning training memory test

Incubation period (S) error number latency (S) error number sham operation group 28.69±9.74 1.90±0.86 212.32±19.56 1.52±0.46 model group 49.38±7.32** 7.62±1.78** 87.96±18.64** 7.20±2.18** Medicine group 1 30.46±5.28 ^## 3.98±1.24 ^## 146.94±22.40 ^## 2.70±1.87 ^## Note: **P < 0.01 , ~~ ##P < 0.01

As can be seen from Table 11, the administration group 1 was compared with the model group ^## P < 0.01, indicating that the composition of the present invention can significantly shorten the incubation period of the learning training and the memory test, and reduce the number of errors, and it can be seen that the composition of the present invention can improve vascular dementia. Rat's ability to learn and remember.

 Conclusion: The composition of the present invention can significantly improve the learning and memory ability of the rats with bilateral common carotid vascular dementia by permanent ligation, and thus has a protective effect on rats with vascular dementia. Example 5 Experimental study on the protective effect of the composition of the present invention on stress and depression animals

 Wistar rats were incubated in a quiet, warm environment for 1 week before the experiment. All the animals were single-cage, except for the control group. They were naturally light, well ventilated, and free to eat water.

 Chronic mild unpredictable stress (CUMS) depression model production plan: 2h behavior limitation, 4V ice swimming for 5min, water stop 24h, food 24h, tail lmin, light and dark reverse 24h, electric shock foot 5s (30V voltage Nine kinds of stimuli, such as shaking for 5min (160Hz) and 40°C for 5min, are randomly arranged within 18 days, one per day, so that the animals can not predict the occurrence of irritation. Wistar rats were randomly divided into normal control group, depression model group, fluoxetine positive control group, Chinese herbal extract administration group 1, 2, 3 (20 g crude drug/kg, 10 g crude drug/kg and 5 g crude drug/kg). The administration method was the same as in Example 2) for 21 days, and each group of animals was decapitated after 24 hours of the last administration.

 Experimental index determination

 Behavioral Determination - Morris Water Maze Test

 The training and testing of spatial memory was carried out in accordance with the Morris water Maze method. The water temperature is maintained at around 23 °C and the water is dyed white to conceal the platform. The surrounding environmental conditions remained quiet and stable during the experiment.

 The experiment is divided into training period and testing period, and l-4d is the training period. During the training, the rats were placed in the water against the platform, and the escape latency of the rats was recorded, that is, the time required from the time the rats entered the water to find the underwater hidden platform. If the platform was not found within 60s, the rats were guided to find the platform. . The escape latency is measured in 60s. The rats are found or guided to the platform for 20s, then dried and returned to the cage. Each rat's water entry position is changed clockwise, with each training interval being 30 s. The next day's training is one quadrant into the water than the first day, and so on. The 5th and 7th are the test period, and the platform is removed during the test. Both tests recorded the platform-to-time ratio of the rats, which is the ratio of the time in which the rats in the quadrant of the original platform traveled within 60 seconds of the total time (60s).

One-Way ANOVA analysis, Ρ<0.05 was considered statistically significant. The experimental results are shown in Table 12.

Table 12 Average escape latency of each group of rats during training period (±sd, n=12)

Average escape latency (s) Normal control group 22.1+±4.3

 Depression model group 34.9±4.8

Fluoxetine positive control group 26.5±3.7 ^#

Drug administration group 1 26.2±4.5 ^#

 Administration group 2 26.7±6.1

Administered group 3 27.5 ± 3.2 ^{# Note: P <0.01, # P <} 0.05

 As can be seen from Table 12, the drug-administered group was compared with the model group: #P<0.05, indicating that the rat's escape latency was significantly shortened after administration of the composition of the present invention, that is, the composition of the present invention can improve the learning and memory ability of the stress-induced depression rats. .

 Determination of biochemical indicators - determination of monoamine oxidase (MAO), glutathione S-transferase (GST), gamma glutamyl transferase (γ-GT)

 After the last treatment of Wistar rats, the rats were decapitated and the cerebral cortex of the rats was rapidly separated on the ice tray, and the lower brain stem was removed. After weighing, 0.86% pre-cooled physiological saline 1:9 was homogenized in an ice water bath to make a 10% brain homogenate. After each sample was dispensed in several portions, the refrigerator was stored in a low temperature freezer. The prepared homogenate was centrifuged at 3000 r/min, and centrifuged for 15 minutes to obtain a homogenate supernatant, and the enzyme activities of MAO, GST, and γ-GT were respectively determined by a chromogenic substrate method according to the method described in the kit. The results are shown in Table 13.

 Table 13 Results of biochemical indicators of rat brain homogenate (;^ ±sd, n=12)

 MAO activity GST activity γ-GT activity group

μ/g protein μ/g protein μ/g protein normal control group 20.35 ±9.23 18.79±4.78 87.66±20.45 depression model group 35.24±7.45 33.76±6.33** 59.64±17.21 "fluoxetine positive control group 21.87±9.64 ** 20.04±8.34 Ding 76.13±22.75 Pu-administered group 1 22.19±7.73 Pu 22.15±5.82 Pu 79.65±18.39 ^## Administration group 2 24.38±9.67 Pu 25.83±5.46 Pu 74.37±20.15 ^#药物组3 26.02±7.58 ^# 27.01 ±3.98 ^# 72.97±23.64 ^#

0.01, ^ff P<0.05, ^ffff P<0.01

As apparent from Table 13, compare treatment group and the model ^{group: # P <0.05, P <} 0.01, compositions of the present invention can be significantly improved monoamine oxidase (of MAO), glutathione S - transferase (GST), γ glutamyl Transferase (γ-GT) activity. Conclusion: The composition of the present invention has a certain protective effect on chronic stress-induced depression rats. Example 6 Experimental study on the protective effect of the composition of the present invention on bacterial meningitis

 Establishment of a bacterial meningitis model

 Experimental animal

 Because bacterial meningitis has a high incidence in infants and young children and is a serious acute infectious disease in the neonatal period, the experimental animals of bacterial meningitis use 11-day-old SD newborn rats.

 Pathogenic bacteria: GBSIII strains were purchased from China National Institute for the Control of Pharmaceutical and Biological Products.

The dried strains were cultured in serum broth for 24 hours at 37 ° C, and then streaked into blood plates. After 18-24 hours, a single colony was selected into serum broth medium to collect in the middle of logarithmic growth. The OD value corresponding to the adjusted turbid liquid concentration by the turbidimetric method was the same as that of the 0.5 standard turbidity tube (concentration lx l0 ⁸ cfu/ml), and the bacterial liquid concentration was 10 ⁸ cfu/ml. Dilute the bacteria to the desired concentration of 10 ⁴ cfu/ml with physiological saline.

 1) Experimental group SD neonatal rats were anesthetized by intraperitoneal injection of 1.0% sodium pentobarbital (15-25 mg/kg). The lOul puncture needle was placed vertically in the 2-4 mm depression (cerebellar medullary pool) at the midpoint of the midpoint of the two ear tips of the newborn rats. After a clear breakthrough, the clear cerebrospinal fluid (CSF) flowed out and the l OulCSF was discarded. . The experimental group was slowly injected with 10 ul of GSBIII suspension. The whole process was completed in 1 and 2 minutes. After surgery, the patient was placed at a room temperature of 25 °C. After resuscitation, the rats were returned to the cage and fed by the mother. After culturing for a certain period of time, the experimental group CSF is extracted and processed into CSF stock solution, cefotaxime + CSF, Chinese herbal medicine extract 20g crude drug / kg + CSF, Chinese herbal medicine extract 10g crude drug / kg + CSF, Chinese herbal medicine extract of the present invention 5 g of crude drug/kg + CSF several different groups of brain infusion.

 2) Fresh SD rats were randomly divided into: blank control group, model group, positive control group, and drug-administered group 1, 2, 3. Anesthesia was administered intraperitoneally with 1.0% sodium pentobarbital (15-25 mg/kg). Holding a 10ul puncture needle at the midpoint of the midpoint of the two ear tips of the newborn rats, the 2-4mm depression (cerebellar medullary pool) was inserted vertically, and after a clear breakthrough, the clear cerebrospinal fluid (CSF) flowed out, and the OulCSF was taken out. Injecting the same amount of CSF stock solution, cefotaxime + CSF, 20 g of crude drug/kg + CSF (administering group 1), and 10 g of crude drug/kg + CSF (administering group 2) into the respective groups, 5 g crude drug / kg + CSF (administration group 3). Postoperative supine, placed at room temperature 25 °C, after resuscitation, put back into the cage and continue feeding.

 Clinical symptom observation

 After inoculation of the bacteria, clinical observations include: presence or absence of cleft and limb skin blemishes, difficulty breathing, unresponsiveness, decreased movement and convulsions, and the time at which symptoms first appear. Neonatal rats were evaluated 24 hours after inoculation with Loeffler's neurobehavioral 5-point scale.

5 points: When grabbing the back, you can move normally, turn over within 5s; 4 points, reduce spontaneous movement, turn over within 5s; 3 points: 〉5s turn over; 2 points: Can't turn over; 1 point: Can't move. For the occurrence of convulsions, the degree of convulsion was evaluated according to Racine's sputum grading criteria for SD rats. The standard is divided into 6 levels, level 0: no convulsions; level I: facial fascia; level II: foundation at level I On, there is a rhythm nod; Class III: There are forelimb clumps on the basis of Class II; Class IV: There are hind leg standing forces on the basis of Class III; Class V: There is a fall on the basis of Class IV.

 The experimental results are shown in Tables 14 and 15.

 Table 14 Clinical manifestations of neonatal rats vaccinated with different fluids group n snoring dyspnea dyskinesia convulsions blank control group 100 000 model group 10 9 9 7 6 positive control group 10 2 1 2 1 drug group 1 10 2 2 2 2 Administration group 2 10 3 4 2 3 Administration group 3 10 3 5 3 3 As apparent from Table 14, the clinical symptoms of the rats were significantly alleviated after administration of the composition of the present invention.

 Table 15 Inoculation of different fluids 24h neurobehavioral score comparison group n score (±s)

 Blank control group 10 5±0

 Model group 10 2.87±1.09

Positive control group 10 3.99±0.87 ^##

 Drug administration group 1 10 4.27±1.12

Drug administration group 2 10 3.79±1.04 ^##

Administration group 3 10 3.43±1.36 ^#

0.01, ^ff P < 0.05, ^ffff P < 0.01

As can be seen from Table 15, one-Way ANOVA analysis, the model group was compared with the blank control group: "Ρ < 0.01 indicates successful modeling; the drug-administered group compared with the model group: #Ρ<0.05, ^## Ρ <0.01, indicating that the neurological behavior of the drug-administered group was significantly improved compared with the model group, and it can be seen that the composition of the present invention can significantly improve the neurobehavior of meningitis rats.

 Rats that survived after 24 h of inoculation were sacrificed by cerebellar medullary puncture and counted as OBC CSF for WBC (white blood cell calculation) before counting. The results are shown in the following table 16:

 Table 16 WBC calculation results

 Group n mean (±s)

 Blank control group 5 13.8±1.23

Model group 5 14780.0±5978** positive control group 5 9920±1745 ^#

 Administration group 1 5 9054±2346

 Drug administration group 2 5 9847±3890

Administration group 3 5 10976±4577 ^#

0.01, ^ff P < 0.05, ^ffff P < 0.01 From Table 16, one-Way ANOVA analysis showed that the model group was compared with the blank control group: "P < 0.01, indicating successful modeling; the drug-administered group compared with the model group: #P<0.05, ## P < 0.01, indicating that the composition of the present invention can significantly reduce the number of white blood cells in rats and reduce the inflammation in rats with meningitis.

 Conclusion: The composition of the present invention has a good antibacterial effect and has a certain improvement effect on the symptoms of bacterial meningitis animals.

Claims

Claims:

1. A composition of a traditional Chinese medicine compound or extract, the traditional Chinese medicine consisting of ginseng, berberine and medlar.

The composition of a traditional Chinese medicine compound or extract according to claim 1, wherein the amount of the ginseng, berberine, and medlar is 100 parts by weight: ginseng, 1-99 parts, berberine, 1 -99 parts, hazelnut, 1-99 parts; preferably ginseng, 2-75 parts, berberine, 2-75 parts, wolfberry, 2-75 parts; more preferably ginseng 3-50 parts, berberine, 3-50 parts, Scorpion, 3-50 servings.

3. The composition of a traditional Chinese medicine compound or extract according to claim 1, wherein the content of the ginseng extract, the extract of Coptidis Rhizome, and the extract of Gardenia is calculated according to the total weight of 100 parts: ginseng extract, 1- 99 parts, Coptidis Rhizoma extract, 1-99 parts, Gardenia extract, 1-99 parts; Preferred ginseng extract, 2-75 parts, Coptis extract, 2-75 parts, Gardenia extract, 2-75 parts More preferably, ginseng extract 3-50 parts, Coptidis Rhizoma extract, 3-50 parts, Gardenia extract, 3-50 parts.

The composition of a traditional Chinese medicine compound or extract according to claim 2, wherein the weight ratio of the ginseng, berberine, and medlar is 3:2: 2-0.5, preferably 3:2: 2-1, More preferably, it is 3:2:2.

The composition of the traditional Chinese medicine compound or extract according to claim 3, characterized in that the weight ratio of the ginseng extract, the coptis extract, the hazelnut extract is 3:2: 2-0.5, preferably 3: 2:2-1, more preferably 3:2:2.

6. A pharmaceutical preparation comprising the composition of claim 1 as an active ingredient and any one or more pharmaceutically acceptable carriers.

7. A pharmaceutical formulation according to claim 6 which is in any pharmaceutically acceptable dosage form.

8. Use of a composition according to claim 1 for the manufacture of a medicament for the treatment or prevention of brain and nerve damage, neurodegenerative encephalopathy, bacterial or viral encephalitis, or depression.

9. The use according to claim 8, wherein the brain and nerve damage is cerebrovascular dementia or ischemic cerebrovascular disease.

10. The use according to claim 9, wherein the ischemic cerebrovascular disease is a transient local brain Ischemic attack, cerebral infarction or stroke.

The use according to claim 8, wherein the neurodegenerative encephalopathy is Alzheimer's disease or Parkinson's disease.