Preparation method of traditional Chinese medicine component and traditional Chinese medicine composition for treating senile dementia

Abstract

The invention provides a preparation method of a traditional Chinese medicine component and a traditional Chinese medicine composition for treating senile dementia, and belongs to the technical field of medicine preparation.The preparation method comprises the steps that sodium chloride and glucose are taken and added into a Tris-HCl buffer solution to be dissolved, a safflower medicinal material and acid cellulase are added into an obtained Tris solvent, ultrasonic enzymolysis is conducted, after enzymolysis is completed, an oil body is collected, and the traditional Chinese medicine composition for treating senile dementia is obtained. The safflower oil is obtained. The raw materials of the traditional Chinese medicine composition for treating senile dementia comprise the prepared safflower oil, borneol, vitamin E and vitamin B6. According to the method, the acidic cellulase is matched with ultrasonic hydrolysis to break and remove safflower cell walls, so that the safflower oil is separated out from safflower cells and dispersed into the Tris solvent, the stability of the safflower oil is maintained through sodium chloride and glucose dissolved in the Tris solvent, and the recovery rate of the safflower oil and the content of gamma-linolenic acid and linoleic acid in the safflower oil can be increased.

Description

Technical Field

[0001] The invention relates to a method for preparing Chinese medicinal materials, and in particular to a method for preparing Chinese medicinal components and a Chinese medicinal composition for treating Alzheimer's disease. Background Art

[0002] Alzheimer's disease is a progressive cognitive dysfunction syndrome caused by damage to brain nerve cells. Its main feature is the gradual decline of cognitive functions, including memory, language, thinking, orientation, understanding, calculation, learning ability and other aspects. The decline of these abilities will seriously affect the patient's daily life and social activities. Alzheimer's disease includes three types: Alzheimer's disease (AD), vascular dementia (VaD) and Lewy body dementia (DLB).

[0003] The pathological characteristics of Alzheimer's disease (AD) are mainly the appearance of senile plaques formed by the deposition of β-amyloid protein (Aβ) and neurofibrillary tangles (NFTs) in the brain. The accumulation of these abnormal substances between nerve cells can cause neuronal degeneration and necrosis, which in turn affects the transmission of nerve signals. For example, in the hippocampus of the brain (a brain area closely related to memory), damage to neurons can cause patients to have early memory impairment, such as forgetting what just happened and asking the same questions repeatedly.

[0004] Vascular dementia (VaD) is usually caused by cerebrovascular diseases, such as cerebral infarction, cerebral hemorrhage, etc. When cerebral blood vessels are damaged, the blood supply to the local part of the brain is affected, resulting in hypoxia and ischemia of brain tissue and damage. Its symptoms are related to the location and degree of cerebrovascular disease.

[0005] Lewy bodies appear in the brain neurons of patients with Lewy body dementia (DLB). Its clinical manifestations are relatively complex. Patients not only have cognitive impairment, but are often accompanied by Parkinson's-like movement symptoms, such as limb stiffness, tremor, and bradykinesia. In addition, patients' cognitive functions fluctuate greatly, and they may be sometimes clear and sometimes confused within a day or several days, and may also experience vivid hallucinations.

[0006] Chinese invention patent application number 200410092423.7 discloses a composition for treating dementia, which has the effects of prolonging the platform jumping latency of AD rats, reducing the number of errors, improving learning and memory, inhibiting MAO activity and reducing the decomposition of monoamine neurotransmitters, promoting learning and memory functions, and enhancing stress sensitivity. However, the efficacy of this Chinese medicine composition needs to be further improved.

[0007] It can be seen from the above patent that Xinnaoqing made from safflower oil, borneol, vitamin E and vitamin B6 has a good therapeutic effect on Alzheimer's disease, but the efficacy needs to be further improved. Summary of the invention

[0008] In view of the above problems, the present invention provides a method for preparing a Chinese medicine component and a Chinese medicine composition for treating Alzheimer's disease.

[0009] To achieve the above purpose, the technical solution adopted by the present invention is:

[0010] A method for preparing a traditional Chinese medicine component comprises the following steps: adding sodium chloride and glucose to a Tris-HCl buffer for dissolution, adding safflower medicinal materials and acid cellulase to the obtained Tris solvent, performing ultrasonic enzymolysis, and collecting oil bodies after the enzymolysis is completed to obtain the safflower oil.

[0011] Furthermore, the content of sodium chloride in the Tris solvent is 0.175-0.19 g / L, and the content of glucose is 0.11-0.125 g / L.

[0012] Furthermore, the pH value of the Tris-HCl buffer is 7.0-7.2.

[0013] Furthermore, the weight-to-volume ratio of safflower medicinal material to Tris solvent is 1kg:9.5-10.5L.

[0014] Furthermore, the weight ratio of safflower medicinal material to acid cellulase is 1:0.013-0.02.

[0015] Furthermore, the temperature of ultrasonic enzymolysis is 50-55°C, the ultrasonic frequency is 300-350W, and the time is 30-60min.

[0016] Further, the specific process of collecting oil bodies is as follows:

[0017] After the ultrasonic enzymatic hydrolysis is completed, the obtained system is centrifuged at 8000-10000 rpm and 0-4°C for 10-20 min to remove impurities and collect the upper milky substance;

[0018] The upper layer of emulsion is added to the Tris solvent, stirred and dispersed for 30 to 60 minutes, and then centrifuged at 8000 to 10000 rpm and 0 to 4°C for 10 to 20 minutes to remove impurities, and the upper layer of emulsion is collected for the second time; wherein the volume weight ratio of the Tris solvent to the safflower medicinal material is 4.7 to 5.2 L: 1 kg;

[0019] The upper emulsion collected for the second time is added to the Tris solvent again, stirred and dispersed for 30 to 60 minutes, and then centrifuged at 8000 to 10000 rpm and 0 to 4°C for 10 to 20 minutes to remove impurities and collect safflower oil bodies to obtain the safflower oil; wherein the volume weight ratio of the Tris solvent to the safflower medicinal material is 4.7 to 5.2 L: 1 kg.

[0020] Furthermore, the preparation method comprises the following specific steps:

[0021] S1. Grind safflower seeds and pass them through a 30-50 mesh sieve to obtain the crushed safflower medicinal material.

[0022] S2, taking Tris-HCl buffer, adding sodium chloride and glucose to dissolve, and obtaining Tris solvent;

[0023] Add the crushed safflower medicinal material to a Tris solvent, stir, then add acid cellulase, heat to 50-55° C., maintain 50-55° C. for ultrasonic enzymolysis for 30-60 min, and the ultrasonic frequency is 300-350 W. After the enzymolysis is completed, centrifuge at 8000-10000 rpm and 0-4° C. for 10-20 min to remove impurities and collect the upper milky substance;

[0024] S3, add the upper emulsion to 4.7-5.2 L of Tris solvent, stir and disperse for 30-60 min, then centrifuge at 8000-10000 rpm and 0-4°C for 10-20 min to remove impurities, and collect the upper emulsion for the second time;

[0025] S4. The upper emulsion collected for the second time is added to 4.7-5.2 L of Tris solvent again, stirred and dispersed for 30-60 min, and then centrifuged at 8000-10000 rpm and 0-4° C. for 10-20 min to remove impurities and collect safflower oil bodies to obtain the safflower oil.

[0026] A Chinese medicine composition for treating Alzheimer's disease, wherein the raw materials for making the effective ingredients of the Chinese medicine composition for treating Alzheimer's disease include, by weight: 390 parts of safflower oil prepared by the above preparation method, 3 parts of borneol, 17 parts of vitamin E and 5 parts of vitamin B6.

[0027] Furthermore, the preparation method of the Chinese medicine composition for treating Alzheimer's disease is to mix all raw materials, and then obtain the soft capsules by shaping, washing and drying.

[0028] The beneficial effects of the preparation method of the Chinese medicine component and the Chinese medicine composition for treating Alzheimer's disease of the present invention are:

[0029] The invention can fix the γ-linolenic acid and linoleic acid in the extracted safflower oil at a specific ratio through a specific extraction process, and can increase the content thereof, thereby improving the efficacy of the prepared traditional Chinese medicine composition for treating Alzheimer's disease;

[0030] The invention utilizes acid cellulase in combination with ultrasonic hydrolysis to break safflower cell walls, so that safflower oil is precipitated from safflower cells and dispersed in a Tris solvent, and the stability of the safflower oil is maintained by sodium chloride and glucose dissolved in the Tris solvent, which is beneficial to improving the recovery rate of safflower oil and the contents of gamma-linolenic acid and linoleic acid in the safflower oil;

[0031] A specific amount of sodium chloride in the Tris solvent can change the ion concentration, which helps stabilize the surface charge of safflower oil bodies, so that the safflower oil bodies are under a certain electric field, which is conducive to the dispersion of the oil bodies, and a specific salt concentration can also help suspend the safflower oil bodies;

[0032] A specific amount of glucose in the Tris solvent can help maintain the stability of the oil bodies, prevent the oil bodies from agglomerating or deteriorating, and prevent the safflower oil from being oxidized, acting as a stabilizer and protective agent; a specific amount of glucose can make the safflower oil bodies disperse more evenly;

[0033] At the same time, a specific amount of glucose at a specific temperature can also promote cellulase hydrolysis and accelerate the release of safflower oil bodies from cells;

[0034] When the pH value of Tris solvent is 7.0-7.2, it can ensure the stability of safflower oil body potential, improve the activity of cellulase, and thus increase the extraction rate of safflower oil.

[0035] The safflower oil extraction process of the invention is simple and easy to industrialize; the obtained safflower oil has uniform and stable oil body and good quality. DETAILED DESCRIPTION

[0036] The technical solutions in the embodiments of the present invention are described clearly and completely below. In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention can also be implemented in other ways different from those described herein, and those skilled in the art can make similar generalizations without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0037] Example 1 A method for preparing a Chinese medicine component

[0038] This embodiment is a method for preparing a Chinese medicine component, and the specific preparation process includes the following steps performed in sequence:

[0039] S1. Grind safflower seeds and pass them through a 30-mesh sieve to obtain pulverized safflower medicinal material (oil content: 24.8%).

[0040] S2, take 20L of Tris-HCl buffer with a pH of 7.0, add 3.8g of NaCl and 2.2g of glucose and dissolve them to obtain Tris solvent;

[0041] Take 1 kg of crushed safflower medicinal material, add it to 10 L of Tris solvent, stir, then add 15 g of acid cellulase (suitable for pH = 7 environment), heat to 50°C, maintain 50°C ultrasonic enzymatic reaction for 30 minutes, the ultrasonic frequency is 300W, after the enzymatic reaction is completed, centrifuge at 8000 rpm and 4°C for 10 minutes to remove impurities and collect the upper milky substance; wherein the acid cellulase is any cellulase suitable for a neutral environment.

[0042] S3, add the upper emulsion to 5L of Tris solvent, stir and disperse for 30min, then centrifuge at 8000rpm and 4℃ for 10min to remove impurities, and collect the upper emulsion for the second time;

[0043] S4. The upper emulsion collected for the second time was added to 5 L of Tris solvent again, stirred and dispersed for 30 min, and then centrifuged at 8000 rpm and 4 ° C for 10 min to remove impurities and collect safflower oil bodies to obtain 0.203 kg of safflower oil (marked as M1), with an oil body yield of 20.3% (recovery rate of 76.35%), a water content of 6.73%, a linoleic acid content of 71.1%, and a γ-linolenic acid content of 13.6%.

[0044] Among them, the content of γ-linolenic acid was determined according to "Determination of γ-linolenic acid content in safflower oil soft capsules by gas chromatography" by Chen Aiying et al.

[0045] The method for determining the linoleic acid content is as follows:

[0046] Determination of correction factor: Take an appropriate amount of n-tricosane, weigh it accurately, and add n-hexane to make a solution containing 4 mg per 1 mL as the internal standard solution. Take about 10 mg of linoleic acid methyl ester reference substance, weigh it accurately, put it in a 10 mL volumetric flask, accurately add 2 mL of internal standard solution, add n-hexane to the scale, and shake well. Take 1uL, inject it into the gas chromatograph, and calculate the correction factor (linoleic acid weight = linoleic acid methyl ester weight × 0.9524).

[0047] Determination method: Take about 35 mg of refined safflower oil, accurately weigh, place in a stoppered test tube, add 2 mL of 0.5 mol / L potassium hydroxide methanol solution, saponify in a 60°C water bath for 10 minutes, cool, add 2 mL of 15% boron trifluoride methanol solution, methylate in a 60°C water bath for 2 minutes, cool, accurately add 5 mL of n-hexane, shake to extract, add 2 mL of saturated sodium chloride solution, shake well, let stand, accurately measure 2 mL of supernatant, place in a 10 mL volumetric flask, accurately add 2 mL of internal standard solution, add n-hexane to the scale, shake well. Take 1 μL and inject it into the gas chromatograph, determine according to the gas chromatography method (General Rule 0521), and calculate (weight of linoleic acid = weight of linoleic acid methyl ester × 0.9524).

[0048] Chromatographic conditions: using succinic acid dioxalate polyester (DEGS) as the stationary phase, the coating concentration is 15%, the column temperature is 180°C, and the theoretical plate number calculated based on the linoleic acid methyl ester peak should not be less than 1500.

[0049] Preparation methods of Chinese medicine components of embodiments 2 to 6

[0050] Examples 2 to 6 are respectively methods for preparing a Chinese medicine component, and their steps are basically the same as those of Example 1, except that the amount of raw materials used and the process parameters are different, as shown in Table 1:

[0051] Table 1 List of process parameters in Examples 2 to 6

[0052]

[0053] The process parameters and steps of other parts of Examples 2 to 6 are the same as those of Example 1.

[0054] Example 7 Preparation method of a Chinese medicine composition for treating Alzheimer's disease

[0055] This embodiment is a method for preparing a Chinese medicine composition for treating Alzheimer's disease. The specific preparation process includes the following steps performed in sequence:

[0056] Take 390g of safflower oil M1 prepared in Example 1, add 3g of borneol, 17g of vitamin E, and 5g of vitamin B6, grind and mix them together, stir evenly, pass through a 100-mesh sieve, press into soft capsules, and then shape, wash and dry them to obtain a traditional Chinese medicine composition for treating Alzheimer's disease, marked as N1.

[0057] Examples 8 to 12 Preparation of a Chinese medicine composition for treating Alzheimer's disease

[0058] Examples 8 to 12 are respectively methods for preparing a Chinese medicine composition for treating Alzheimer's disease, and their steps are basically the same as those of Example 7, except that different safflower oils are used:

[0059] In Example 8, the safflower oil M2 refined in Example 2 is used, and the obtained Chinese medicine composition for treating Alzheimer's disease is marked as N2;

[0060] The safflower oil M3 refined in Example 3 was used in Example 9, and the obtained Chinese medicine composition for treating Alzheimer's disease was marked as N3;

[0061] In Example 10, the safflower oil M4 refined in Example 4 is used, and the obtained Chinese medicine composition for treating Alzheimer's disease is marked as N4;

[0062] In Example 11, the safflower oil M5 refined in Example 5 is used, and the obtained Chinese medicine composition for treating Alzheimer's disease is marked as N5;

[0063] The safflower oil M6 refined in Example 6 is used in Example 12, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease is marked as N6.

[0064] Experimental Example 1 Preparation method of Chinese medicine components

[0065] Comparative Examples 1 to 6 are comparative tests of the preparation method of the Chinese medicinal components in Example 1, and the same batch of safflower seeds as in Example 1 are used as raw materials. The steps are basically the same as in Example 1, except that:

[0066] In step S2 of comparative example 1, 2.2 g of sucrose was used instead of 2.2 g of glucose, and the amounts of other raw materials, process steps and parameters remained unchanged, to obtain 0.187 kg of safflower oil, labeled as DM1, with an oil yield of 18.7% (recovery rate of 70.31%), a water content of 6.75%, a linoleic acid content of 69.6%, and a γ-linolenic acid content of 11.5%.

[0067] In step S2 of comparative example 2, 4 g of glucose was used instead of 2.2 g of glucose, and the amounts of other raw materials, process steps and parameters remained unchanged, to obtain 0.192 kg of safflower oil, labeled as DM2, with an oil yield of 18.7% (recovery rate of 71.47%), a water content of 7.69%, a linoleic acid content of 65.7%, and a γ-linolenic acid content of 13.4%.

[0068] In step S2 of comparative example 3, 0.5 g of glucose was used instead of 2.2 g of glucose, and the amounts of other raw materials, process steps and parameters remained unchanged, to obtain 0.171 kg of safflower oil, labeled as DM3, with an oil yield of 17.1% (recovery rate of 64.57%), a water content of 6.35%, a linoleic acid content of 70.9%, and a γ-linolenic acid content of 9.3%.

[0069] In step S2 of comparative example 4, 8 g of sodium chloride was used instead of 3.8 g of sodium chloride, and the amounts of other raw materials, process steps and parameters remained unchanged, to obtain 0.168 kg of safflower oil, labeled as DM4, with an oil yield of 16.8% (recovery rate of 63.35%), a water content of 6.49%, a linoleic acid content of 67.9%, and a γ-linolenic acid content of 12.1%.

[0070] In step S2 of comparative example 5, 2 g of sodium chloride was used instead of 3.8 g of sodium chloride, and the amounts of other raw materials, process steps and parameters remained unchanged, to obtain 0.153 kg of safflower oil, labeled as DM5, with an oil yield of 15.3% (recovery rate of 56.77%), a water content of 7.98%, a linoleic acid content of 69.7%, and a γ-linolenic acid content of 8.4%.

[0071] In step S2 of comparative example 6, the temperature of ultrasonic enzymolysis is 30° C. and the ultrasonic frequency is 500 W. The amount of other raw materials, process steps and parameters remain unchanged, and 0.139 kg of safflower oil is obtained, which is marked as DM6, with an oil yield of 13.9% (recovery rate of 52.29%), a water content of 6.71%, a linoleic acid content of 70.8%, and a γ-linolenic acid content of 13.4%. It can be seen that changing the process parameters will lead to a lower overall recovery rate of safflower oil, which is not conducive to production.

[0072] By comparing Example 1 with Comparative Examples 1 to 6, it can be seen that the present invention can effectively improve the recovery rate of safflower oil and increase the contents of linoleic acid and γ-linolenic acid in safflower oil only with specific process parameters.

[0073] Experimental Example 2 Determination of the efficacy of a Chinese medicine composition for treating Alzheimer's disease

[0074] Comparative Examples 7 to 12 are comparative tests of the preparation process of the Chinese medicine composition for treating Alzheimer's disease in Example 7, the only difference being that:

[0075] In Comparative Example 7, the safflower oil DM1 prepared in Comparative Example 1 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN1;

[0076] In Comparative Example 8, the safflower oil DM2 prepared in Comparative Example 2 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN2;

[0077] In Comparative Example 9, the safflower oil DM3 prepared in Comparative Example 3 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN3;

[0078] In Comparative Example 10, safflower oil DM4 prepared in Comparative Example 4 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN4.

[0079] In Comparative Example 11, safflower oil DM5 prepared in Comparative Example 5 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN5.

[0080] In Comparative Example 12, safflower oil DM6 prepared in Comparative Example 6 was used as a raw material, and the obtained traditional Chinese medicine composition for treating Alzheimer's disease was marked as DN6.

[0081] The Chinese medicinal compositions N1 and DN1 to DN6 for treating Alzheimer's disease prepared in Example 7 and Comparative Examples 7 to 12 were compared for efficacy, as follows:

[0082] 1. Behavioral influence experiment

[0083] 110 healthy male SD rats, weighing 150±17g, were randomly divided into blank group, sham operation group, model group, experimental groups 1 to 7, and control group, with 10 rats in each group. Except for the blank group, rats in other groups were anesthetized with 10% chloral hydrate intraperitoneal injection (3mL / kg), and 1μL of β-AP was injected into the hippocampus according to the stereotaxic atlas (AP=2.22mm, ML-=3.0mm, DR=2.8mm). The sham operation group was injected with an equal amount of saline without β-AP, and the blank group was not treated. Behavioral tests were performed 1 week after surgery. After the test, the drugs were administered at 15 times the human dosage per kilogram, wherein the blank group, sham operation group and model group were not administered, the experimental group 1 was administered with the Chinese medicine composition N1 for treating Alzheimer's disease prepared in Example 1, the experimental group 2 was administered with the Chinese medicine composition DN1 for treating Alzheimer's disease prepared in Comparative Example 7, the experimental group 3 was administered with the Chinese medicine composition DN2 for treating Alzheimer's disease prepared in Comparative Example 8, the experimental group 4 was administered with the Chinese medicine composition DN3 for treating Alzheimer's disease prepared in Comparative Example 9, the experimental group 5 was administered with the Chinese medicine composition DN4 for treating Alzheimer's disease prepared in Comparative Example 10, the experimental group 6 was administered with the Chinese medicine composition DN5 for treating Alzheimer's disease prepared in Comparative Example 11, the experimental group 7 was administered with the Chinese medicine composition DN6 for treating Alzheimer's disease prepared in Comparative Example 12, and the control group was administered with Naofukang (piracetam), and behavioral tests were performed again 3 weeks after administration. Wherein, β-AP was prepared into a solution with a concentration of 5 μg / μL with physiological saline before use.

[0084] Behavioral measurement: The rat jumping platform method was used. The animals were placed in the jumping platform and adapted to the environment for 5 minutes. The time it took for the rats to escape to the safety platform and then jump off the safety platform was the latency period, and the number of electric shocks received within 5 minutes was the number of errors.

[0085] Statistical analysis The measured data were expressed as (x ± s), and the t test was used for the significance test of the mean between groups. The specific results are as follows:

[0086] Table 2 Summary of behavioral (latency) (x ± s) results

[0087]

[0088]

[0089] Table 3 Summary of behavioral (number of errors) (x ± s) results

[0090] Group Before treatment After treatment Blank Group 0.37±0.69 / Sham injury group 2.49±0.73 2.69±0.82 Model Group 1.98±0.89 4.99±1.17 Control group 1.86±0.91 2.67±1.09 Experimental group 1.95±0.97 2.09±0.79 Experimental Group 2 1.87±0.84 2.52±1.05 Experimental Group 3 1.77±0.93 2.66±1.16 Experimental Group 4 1.83±0.69 2.54±0.93 Experimental Group 5 1.92±1.05 2.61±1.21 Experimental Group 6 1.88±0.79 2.58±0.99 Experimental Group 7 1.84±0.96 2.02±1.07

[0091] It can be seen from Tables 3 and 4 that the Chinese herbal compositions N1 and DN6 for treating Alzheimer's disease can prolong the platform jumping latency of AD rats, reduce the number of errors, and improve learning and memory, while the therapeutic effect of the Chinese herbal compositions DN1 to DN5 for treating Alzheimer's disease is poor, indicating that a specific ratio of linoleic acid and γ-linolenic acid content can improve the therapeutic effect on AD rats.

[0092] 2. Experiment on the impact of vascular dementia

[0093] 100 healthy male SD rats weighing 150±17g were randomly divided into a blank group, a model group, experimental groups 1 to 7, and a control group, with 10 rats in each group. Except for the blank group, rats in the other groups were anesthetized with 10% chloral hydrate intraperitoneally (3mL / kg), and bilateral common carotid arteries were permanently ligated. The administration method is that the rats of experimental groups one to seven and the control group are administered drugs at the same time during modeling. The rats in the experimental group are gavaged with 2 mL of the solution of the traditional Chinese medicine composition N1 for treating Alzheimer's disease every day, the rats in the experimental group two are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN1 for treating Alzheimer's disease every day, the rats in the experimental group three are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN2 for treating Alzheimer's disease every day, the rats in the experimental group four are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN3 for treating Alzheimer's disease every day, the rats in the experimental group five are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN4 for treating Alzheimer's disease every day, the rats in the experimental group six are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN5 for treating Alzheimer's disease every day, the rats in the experimental group seven are gavaged with 2 mL of the solution of the traditional Chinese medicine composition DN6 for treating Alzheimer's disease every day, and the rats in the control group are gavaged with 0.5 mg / kg of Xidezhen every day, and the administration is continued for 30 days.

[0094] Material processing and index determination Determination of brain tissue MAO content and SOD activity: Take the right side of the rat brain tissue, weigh it, put it in a micro homogenizer, add 0.1 mL of anhydrous ethanol to grind it, then add 0.9 mL of physiological saline to fully grind it into a homogenate, centrifuge it at 4°C and 4500 rpm for 15 minutes, and take the supernatant. Use the thiobarbituric acid method and xanthine oxidase method respectively.

[0095] Measurement of NO and NOS: Quickly take 1g of left cerebral tissue and homogenize it with physiological saline at a ratio of 0.1g:1mL, centrifuge at 3000rpm for 10min, take the supernatant and place it in a -20℃ refrigerator for testing. Refer to the instructions of the kit. The results are shown in Tables 4 and 5.

[0096] Effect of the soft capsule of the present invention on the activities of MAO and SOD in brain tissue of rats with vascular dementia

[0097] Table 4 Results of MAO and SOD activities (x±s) in brain tissue of rats with vascular dementia

[0098] Group MAO SOD Blank Group 2.54±0.17 205.12±8.96 Model Group 4.05±0.16 125.69±14.28 Control group 3.17±0.14 168.46±11.27 Experimental group 2.76±0.15 189.67±9.69 Experimental Group 2 3.07±0.19 170.24±13.07 Experimental Group 3 2.86±0.15 176.66±12.14 Experimental Group 4 3.03±0.16 172.55±12.85 Experimental Group 5 2.99±0.18 173.96±13.68 Experimental Group 6 2.93±0.19 175.12±11.06 Experimental Group 7 2.79±0.17 188.96±10.76

[0099] The activity of monoamine oxidase (MAO) increases with age, and causes changes in the content of catecholamines in the brain, promoting physiological activity disorders. It is closely related to learning and attention. The activity of MAO in the tissues of aging and AD patients increases, oxidatively decomposing monoamine neurotransmitters, affecting learning and attention, and is one of the causes of senile neurodegenerative diseases. The Chinese medicine compositions N1 and DN6 for treating Alzheimer's disease prepared by the present invention can better inhibit MAO activity, reduce the decomposition of monoamine neurotransmitters, promote learning and memory functions, and enhance stress sensitivity, while the therapeutic effect of the Chinese medicine compositions DN1 to DN5 for treating Alzheimer's disease is poor. Oxygen free radicals (SOD) play an important role in the pathological changes of AD. The Chinese medicine compositions N1 and DN6 for treating Alzheimer's disease prepared by the present invention can scavenge free radicals, promote cell metabolism, and enhance cell vitality, so they can effectively delay cell aging, inhibit cell degeneration and degeneration, improve the structure and function of the body, and improve the vitality of the body, while the therapeutic effect of the Chinese medicine compositions DN1 to DN5 for treating Alzheimer's disease is poor.

[0100] Table 5 Results of NO and NOS activities (x±s) in brain tissue of rats with vascular dementia

[0101] Group NO NOS Blank Group 92.14±24.67 1.20±0.39 Model Group 143.64±30.77 2.03±0.16 Control group 110.12±28.63 1.32±0.27 Experimental group 105.21±26.62 1.25±0.21 Experimental Group 2 109.93±25.96 1.35±0.33 Experimental Group 3 108.25±26.87 1.28±0.29 Experimental Group 4 109.56±31.27 1.33±0.19 Experimental Group 5 109.17±27.96 1.31±0.25 Experimental Group 6 108.84±29.34 1.29±0.28 Experimental Group 7 106.33±27.19 1.26±0.32

[0102] Nitric oxide (NO) is a gaseous substance with a simple structure and extremely unstable, easy to diffuse, highly reactive, short biological half-life, diffused in various brain regions in the central nervous system, and has multiple functions such as regulating cerebral blood flow, participating in synaptic plasticity, neuronal toxicity and inflammatory damage, and is closely related to learning and memory. As can be seen from Table 5, the NO level and NOS activity of the model group continued to increase, indicating that the neurotoxicity of NO participated in the formation of VD. After treatment with the Chinese medicine composition N1 and DN6 for treating Alzheimer's disease prepared by the present invention, the activity of NOS in the cerebral cortex tissue decreased, and the NO generation and release content were significantly reduced, thereby protecting the brain neurons and achieving the purpose of treating VD, while the treatment effect of the Chinese medicine composition DN1 to DN5 for treating Alzheimer's disease was poor.

[0103] Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Claims

1. A method for preparing a Chinese medicine component, characterized in that: The preparation method comprises the following steps: adding sodium chloride and glucose to a Tris-HCl buffer for dissolution, adding safflower medicinal materials and acid cellulase to the obtained Tris solvent, performing ultrasonic enzymolysis, and collecting oil bodies after the enzymolysis is completed, thereby obtaining the safflower oil.

2. The method for preparing the Chinese medicine component according to claim 1, characterized in that: The content of sodium chloride in the Tris solvent is 0.175-0.19 g / L, and the content of glucose is 0.11-0.125 g / L.

3. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The pH value of Tris-HCl buffer is 7.0-7.

2.

4. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The weight-to-volume ratio of safflower medicinal material to Tris solvent is 1kg:9.5-10.5L.

5. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The weight ratio of safflower medicinal material to acid cellulase is 1:0.013-0.

02.

6. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The temperature of ultrasonic enzymatic hydrolysis is 50-55°C, the ultrasonic frequency is 300-350W, and the time is 30-60min.

7. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The specific process of collecting oil bodies is as follows: After the ultrasonic enzymatic hydrolysis is completed, the obtained system is centrifuged to remove impurities and collect the upper milky substance; The upper emulsion was added to Tris solvent, dispersed, and then centrifuged to remove impurities, and the upper emulsion was collected for the second time; The upper emulsion collected for the second time is added to the Tris solvent again, dispersed, and then centrifuged to remove impurities and collect safflower oil bodies to obtain the safflower oil.

8. The method for preparing the Chinese medicine component according to claim 1 or 2, characterized in that: The preparation method comprises the following specific steps: S1. Grind safflower seeds, sieve and obtain the crushed safflower medicinal material. S2, taking Tris-HCl buffer, adding sodium chloride and glucose to dissolve, and obtaining Tris solvent; The crushed safflower medicinal material is added to a Tris solvent, and then acid cellulase is added, heated, and ultrasonic enzymolysis is performed. After the enzymolysis is completed, centrifugation is performed to remove impurities, and the upper layer of milky matter is collected; S3, adding the upper emulsion into Tris solvent, dispersing, then centrifuging to remove impurities, and collecting the upper emulsion for the second time; S4. The upper emulsion collected for the second time is added to the Tris solvent again, dispersed, and then centrifuged to remove impurities and collect safflower oil bodies to obtain the safflower oil.

9. A Chinese medicine composition for treating Alzheimer's disease, characterized in that: In parts by weight, the raw materials for making the effective ingredients of the traditional Chinese medicine composition for treating Alzheimer's disease include: 390 parts of safflower oil prepared by the preparation method described in any one of claims 1 to 8, 3 parts of borneol, 17 parts of vitamin E and 5 parts of vitamin B6.

10. The Chinese medicine composition for treating Alzheimer's disease according to claim 9, characterized in that: The preparation method of the traditional Chinese medicine composition for treating Alzheimer's disease comprises the following steps: mixing all raw materials, and then shaping, washing and drying the obtained soft capsules.

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