Rhizoma cyperi processed product for treating migraine and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine processing, and relates to a rhizoma cyperi processed product for treating migraine and a preparation method thereof. According to the processing technology, a vinegar braising method is adopted, nutgrass galingale rhizome granules and nutgrass galingale rhizome blocks and rice vinegar (3.5-9 DEG C) are fully and uniformly stirred and moistened for 4 hours until vinegar liquid completely permeates into internal tissues of medicinal materials, then the medicinal materials are placed in a marmite to be braised for 3-7 hours, and a finished product is obtained after drying, cooling and impurity removal. According to the invention, the optimal preparation parameters of the vinegar braised rhizoma cyperi are determined for the first time, so that the vinegar braised rhizoma cyperi shows the pharmacological activity which is obviously superior to that of the traditional vinegar rhizoma cyperi in the aspect of treating migraine. Animal experiments prove that the processed product can effectively improve the pain behavior of migraine model rats and remarkably regulate the abnormal level of key neurotransmitters such as 5-HT and CGRP in serum. The invention provides a technical support for developing a novel anti-migraine traditional Chinese medicine with stronger targeting property and clearer curative effect, and has clinical application value and market prospect. The method is clear in technological parameters, simple and convenient to operate and good in repeatability, and the obtained vinegar braised rhizoma cyperi is stable and controllable in quality.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine processing, and particularly relates to a processed product of Cyperus rotundus for treating migraine and a preparation method thereof. Background Art

[0002] Cyperus rotundus is the dried rhizome of the plant Cyperus rotundus L. of the Cyperaceae family, also known as nutgrass flatsedge. Its taste is pungent, slightly bitter, slightly sweet, and its nature is flat. It mainly enters the liver meridian and also enters the spleen and triple energizer meridians. It has the effects of soothing the liver and relieving depression, regulating qi and relieving distension, regulating menstruation and relieving pain. It is a commonly used medicine in traditional Chinese medicine clinical treatment of liver qi stagnation, chest and hypochondrium distending pain, spleen qi stagnation, irregular menstruation and other symptoms, and has the reputation of "the general of qi diseases and the marshal of women's diseases". The traditional processing method is mainly vinegar roasting method. The obtained vinegar-processed Cyperus rotundus can enhance the effect of entering the liver meridian and has the effects of promoting blood circulation and removing stasis, astringing and detoxifying, and relieving pain. However, the traditional processing technology still needs to be further optimized in the precise treatment of specific diseases (such as migraine), and lacks certain modern pharmacodynamic verification.

[0003] Migraine is a common chronic neurovascular disease in clinic. Its pathogenesis is complex and involves abnormal release of neurotransmitters such as 5-hydroxytryptamine (5-HT) and calcitonin gene-related peptide (CGRP). At present, although western medicine treatment has certain effects, long-term use is prone to produce side effects. Although traditional Chinese medicine Cyperus rotundus has certain effects in analgesia, the specific regulatory effect of traditional processed products on migraine is still unclear.

[0004] Therefore, the present invention innovatively proposes a new processing technology of Cyperus rotundus - vinegar stewed Cyperus rotundus. Through the combination of process design, optimization and pharmacodynamic verification, the present invention systematically establishes the preparation method of vinegar stewed Cyperus rotundus and its efficacy in the treatment of migraine for the first time, and has clear scientific basis and application value. Summary of the Invention

[0005] The purpose of the present invention is to provide a processed product of Cyperus rotundus for treating migraine and a preparation method thereof. The preparation method clarifies the processing technology of vinegar stewed Cyperus rotundus and its pharmacodynamic effect in treating migraine, and provides a basis for the research and development of new drugs. The vinegar stewed Cyperus rotundus product obtained by this method has a good effect in treating migraine.

[0006] In order to achieve the above purpose, the present invention adopts the following technical solutions.

[0007] A processed product of Cyperus rotundus for treating migraine is prepared by a vinegar stewing process.

[0008] A preparation process of a processed product of Cyperus rotundus for treating migraine includes the following steps:

[0009] Step 1: Add net Cyperus rotundus granules and pieces to rice vinegar, mix well, and carry out moistening.

[0010] Step 2: Then, it is stewed in a casserole and dried to obtain the processed Cyperus rotundus for treating migraine (vinegar-stewed Cyperus rotundus).

[0011] Furthermore, in Step 1, the mass of the rice vinegar used is 10%-30% of the mass of the net Cyperus rotundus granules and pieces.

[0012] Furthermore, in Step 1, the degree of the rice vinegar is 3.5°-9°.

[0013] Furthermore, in Step 1, the moistening time is 4 h.

[0014] Furthermore, in Step 2, the stewing time in the casserole is 3 h-7 h.

[0015] The vinegar-stewed Cyperus rotundus can be applied in the medicine for treating migraine.

[0016] Compared with the prior art, the present invention has the following beneficial effects.

[0017] In the current existing technology, there is no process for vinegar-stewing Cyperus rotundus, and the existing processed Cyperus rotundus cannot achieve the expected effect of precisely treating migraine. The present invention has carried out key optimizations in the processing technology and found that the amount of added vinegar, the stewing duration, and the acid content in the vinegar are the core factors affecting the medicinal efficacy of vinegar-stewed Cyperus rotundus; among them, the effect of stewing with 10% 3.5° rice vinegar for 3 h is more prominent, which is more beneficial for Cyperus rotundus to treat migraine. By precisely controlling these parameters, its analgesic, anti-inflammatory, and neuromodulatory effects can be enhanced. This process not only fills the blank in the field of Cyperus rotundus processing technology for treating migraine but also provides new ideas for developing traditional Chinese medicines for anti-migraine with higher efficiency, stability, and stronger targeting.

[0018] In this patent, an acute migraine model of rats was established, the behavioral manifestations of rats in each group were observed, and then they were anesthetized to collect blood from the abdominal aorta. 5-HT and CGRP in the serum of rats were detected by ELISA. The experimental results showed that vinegar-stewed Cyperus rotundus could significantly improve the pain behavior of migraine model rats and effectively regulate the abnormal release of 5-HT and CGRP, and its effect was better than that of raw Cyperus rotundus. It provides certain experimental basis for clinical medication. The present invention can improve the medicinal value of the processed Cyperus rotundus, clarify the medicinal efficacy of vinegar-stewed Cyperus rotundus, and provide a basis for further developing Cyperus rotundus into a medicine for treating migraine.

[0019] The preparation method of the vinegar-stewed Cyperus rotundus provided by the present invention has the characteristics of high efficiency, stable process, simple operation, and low cost. Description of the Drawings

[0020] Figure 1 It is the high-performance liquid chromatography diagram of the processed Cyperus rotundus for treating migraine in Example 1. (Among them, 1-5-HMF, 2-ferulic acid, 3-luteoloside, 4-luteolin, 5-cyperenone, 6-nootkatone, 7-α-cyperone) Detailed implementation mode

[0021] The present invention will be further described in detail below with reference to specific embodiments and the accompanying drawings. However, it should not be understood that the scope of the above-mentioned subject matter of the present invention is limited to the following examples. Any technology implemented based on the above content of the present invention belongs to the scope of the present invention.

[0022] A processed product of Cyperus rotundus for treating migraine is prepared by the process of vinegar stewing.

[0023] A preparation process of a processed product of Cyperus rotundus for treating migraine comprises the following steps:

[0024] Step 1: Add the purified Cyperus rotundus granule blocks to 3.5°-9° rice vinegar, mix well, and carry out moistening for 4 h until the rice vinegar is absorbed, so that the rice vinegar completely penetrates into the internal tissue of the cut pieces; the mass of the used rice vinegar is 10%-30% of the mass of the purified Cyperus rotundus granule blocks;

[0025] Step 2: Then use a casserole to stew for 3 h-7 h and then dry to obtain the processed product of Cyperus rotundus for treating migraine (processed product of Cyperus rotundus stewed with vinegar).

[0026] Example 1

[0027] Add the Cyperus rotundus granule blocks to the rice vinegar and mix well. For every 100 kg of Cyperus rotundus, use 20 kg of 6° rice vinegar, moisten for 4 h until it is completely absorbed by the Cyperus rotundus, until the rice vinegar completely penetrates into the internal tissue of the Cyperus rotundus. Place the moistened Cyperus rotundus on a casserole and stew for 3 h; dry, cool, remove impurities, and obtain the processed product of Cyperus rotundus stewed with vinegar.

[0028] Perform chromatographic detection under the chromatographic conditions of Example 3. The chromatogram is shown in Figure 1 .

[0029] Example 2

[0030] Add the Cyperus rotundus granule blocks to the rice vinegar and mix well. For every 100 kg of Cyperus rotundus, use 30 kg of 9° rice vinegar, moisten for 4 h until it is completely absorbed by the Cyperus rotundus, until the rice vinegar completely penetrates into the internal tissue of the Cyperus rotundus. Place the moistened Cyperus rotundus on a casserole and stew for 5 h; dry, cool, remove impurities, and obtain the processed product of Cyperus rotundus stewed with vinegar.

[0031] Example 3

[0032] 1. Instruments and materials

[0033] 1.1 Instruments: Waters e2695 high-performance liquid chromatograph, METTLER AE240 type one-hundred-thousandth analytical balance, numerically controlled ultrasonic cleaner, micropipette (Thermo Fisher), -80°C refrigerator (Qingdao Haier).

[0034] 1.2 Test drugs: Cyperus rotundus herbs were purchased from Kangmei Pharmaceutical Co., Ltd.; 3.5° rice vinegar was purchased from Hunan Changkang Industrial Co., Ltd., 6° rice vinegar was purchased from Sichuan Baoning Vinegar Co., Ltd., and 9° rice vinegar was purchased from Qianhe Flavor & Food Co., Ltd.; methanol was of chromatographic purity, water was purified water, and other reagents were of analytical purity. Reference standards of 5-HMF, ferulic acid, luteoloside, luteolin, cyperenone, nootkatone, and α-cyperone were all purchased from Dalian Meilun Biotechnology Co., Ltd. for content determination, and their purities were all greater than 98%. Adult healthy male SD rats were purchased from Liaoning Changsheng Biotechnology Co., Ltd., nitroglycerin injection was purchased from Beijing Yimin Pharmaceutical Co., Ltd., sumatriptan succinate capsules were purchased from Hunan Jiudian Pharmaceutical Co., Ltd., and rat 5-hydroxytryptamine (5-HT) enzyme-linked immunosorbent assay kit and rat calcitonin gene-related peptide (CGRP) enzyme-linked immunosorbent assay kit were both purchased from Shanghai Kexing Biotechnology Co., Ltd.

[0035] 2. This example provides a preparation method for the vinegar-braised Cyperus rotundus processed product, which includes the following steps:

[0036] Add Cyperus rotundus granules to rice vinegar and mix evenly. For every 100 kg of Cyperus rotundus, use 10 - 30 kg of 3.5° - 9° rice vinegar, moisten for 4 h until completely absorbed by Cyperus rotundus, until the rice vinegar completely penetrates into the internal tissue of Cyperus rotundus, and place the well-moistened Cyperus rotundus on a casserole and braise for 3 h - 7 h; dry, cool, remove impurities, and obtain the vinegar-braised Cyperus rotundus processed product.

[0037] 3. Orthogonal experiment

[0038] Taking the contents of 5-HMF, ferulic acid, luteoloside, luteolin, cyperenone, nootkatone, and α-cyperone after vinegar-braising as evaluation indexes, select the amount of added vinegar, the acid content in the vinegar, and the braising duration as the investigation factors, and use the orthogonal experiment method to optimize the best processing technology for vinegar-braising Cyperus rotundus.

[0039] The factor level table for vinegar-braising Cyperus rotundus in the orthogonal experiment method is designed as follows, see Table 1:

[0040] Table 1 Factor and level table for orthogonal experiment design of vinegar-braising Cyperus rotundus

[0041]

[0042] Determination methods for evaluation indicators of 5-HMF, ferulic acid, luteoloside, luteolin, cyperenone, nootkatone, and α-cyperone: High performance liquid chromatography was used. Chromatographic conditions: Venusil C18 chromatographic column (250 mm × 4.6 mm, 5 μm); methanol - 0.1% phosphoric acid aqueous solution as the mobile phase, with gradient elution: 0 - 12 min, 5% - 20% methanol; 12 - 30 min, 20% - 30% methanol; 30 - 40 min, 30% - 48% methanol; 40 - 52 min, 48% - 50% methanol; 52 - 55 min, 50% - 59% methanol; 55 - 60 min, 59% - 70% methanol; 60 - 80 min, 70% - 77% methanol; detection wavelength: 0 - 65 min, 310 nm; 65 - 80 min, 254 nm; volume flow rate 1.0 mL / min; column temperature 30°C; injection volume 10 μL.

[0043] Solution preparation Preparation of reference solution: Accurately weigh the reference standards of 5-HMF, ferulic acid, luteoloside, luteolin, cyperenone, nootkatone, and α-cyperone separately and place them in 5 mL volumetric flasks. Dilute to the mark with chromatographic methanol to obtain reference solutions with concentrations of 0.102 mg / mL, 0.110 mg / mL, 0.102 mg / mL, 0.090 mg / mL, 0.116 mg / mL, 0.100 mg / mL, and 0.112 mg / mL respectively. Appropriately measure a certain amount of the 7 reference solutions and prepare a mixed reference solution with a certain mass concentration. Preparation of test solution: Crush the sample, accurately weigh an appropriate amount of the sample powder, place it in a stoppered conical flask, accurately add 25 mL of 75% methanol, weigh it, ultrasonically treat it (250 W, 40 kHz) for 45 min, let it cool, make up the lost mass with methanol, shake well, filter, take the subsequent filtrate, and filter it through a 0.22 μm microporous filter membrane to obtain the test solution. The determination results are as follows, shown in Table 2.

[0044] Table 2 Content and comprehensive score of the index components of vinegar-braised Cyperi Rhizoma

[0045]

[0046] Comprehensive score:

[0047] 10×(A i / A max ) + 10×(B i / B max ) + 10×(C i / C max ) + 10×(D i / D max ) + 20×(E i / E max ) + 20×(Fi / F max ) + 20×(G i / G max )

[0048] In the above formula, A is the content of 5-HMF, B is the content of ferulic acid, C is the content of luteoloside, D is the content of luteolin, E is the content of cyperenone, F is the content of nootkatone, G is the content of α-cyperone, and the parameters of the 7 indicators for the comprehensive score of stir-frying Cyperi Rhizoma with vinegar are set to 10, 10, 10, 10, 20, 20, 20 respectively.

[0049] The best technological parameters for stir-frying Cyperi Rhizoma with vinegar are obtained: 10 g of 3.5° rice vinegar is used for moistening 100 g of Rhizoma Cyperi pieces for 4 h, and then it is stewed in a casserole for 3 h. The comprehensive score is 94.10285, and the processing technology of stir-frying Cyperi Rhizoma with vinegar is determined accordingly.

[0050] 4. Experiment on "Treatment of Migraine with Stir-fried Cyperi Rhizoma with Vinegar"

[0051] 4.1 Determination Method

[0052] 4.1.1 Preparation of Administration Solution

[0053] Take an appropriate amount of the above-mentioned stir-fried Cyperi Rhizoma with vinegar, add 10 times the amount of distilled water, soak for 30 min, heat to boiling over high heat and then adjust to simmer for 60 min, stir, filter, and retain the medicinal solution. Add 8 times the amount of distilled water to the medicinal residues, decoct, stir, and filter according to the above method, combine the two medicinal solutions, and concentrate to an appropriate concentration. The concentrations of the administration solutions for the low- and high-dose groups of raw and vinegar Cyperi Rhizoma are 0.10 g / ml and 0.94 g / ml respectively. Take sumatriptan succinate, add an appropriate amount of aqueous solution, and dissolve to a concentration of 25 mg / kg.

[0054] 4.1.2 Grouping, Modeling and Administration

[0055] A total of 42 male SD rats with a body weight between 180 - 220 g are used. After 7 days of adaptive feeding, they are randomly divided into seven groups, including a blank group, a model group, a control group (sumatriptan succinate 25 mg / kg), a low-dose group of stir-fried Cyperi Rhizoma with vinegar (vinegar Cyperi Rhizoma 0.10 g / ml), a high-dose group of stir-fried Cyperi Rhizoma with vinegar (vinegar Cyperi Rhizoma 0.94 g / ml), a low-dose group of raw Cyperi Rhizoma (raw Cyperi Rhizoma 0.10 g / ml), and a high-dose group of raw Cyperi Rhizoma (raw Cyperi Rhizoma 0.94 g / ml). They are continuously gavaged at a dose of 10 mL / kg for 7 days. 30 min after administration on the 7th day, the rats are modeled: an acute migraine model is induced by subcutaneous injection of nitroglycerin (GTN) at 10 mg / kg in the neck (the blank group is water).

[0056] 4.1.3 Observation Indexes and Detection Methods

[0057] Immediately after successful model establishment, observe the scratching head times of the mice within 0 - 30 min; after observing the behavior, intraperitoneally inject 20% urethane at 1 g / kg to anesthetize them, then draw 2 - 5 ml of blood from the abdominal aorta of the rats (note: the blood collection tube should not contain anticoagulant), let it stand at room temperature for 2 h, centrifuge at 4°C and 3000 r / min for 15 minutes, separate the serum, aliquot it, and store it in a -80°C low-temperature refrigerator for later use.

[0058] 4.1.4 Detection Indexes

[0059] Detect 5-HT and CGRP in the rat serum by ELISA.

[0060] 4.1.7 Statistical Methods

[0061] Use the statistical software SPSS to perform statistical analysis on the data. The data are expressed as , and one-way analysis of variance (ANOVA) and LSD test method are used for multiple comparisons between groups. p < 0.05 indicates statistically significant differences.

[0062] 4.2 Measurement Results

[0063] 4.2.1 Effects of Vinegar-Stewed Cyperus rotundus on the Behavior of Migraine Model Rats

[0064] Effects of vinegar-stewed Cyperus rotundus on the behavioral performance of migraine model rats: Compared with the blank group, the scratching head times of the model group, control group, high- and low-dose groups of vinegar-stewed Cyperus rotundus, and high- and low-dose groups of raw Cyperus rotundus were all significantly increased; compared with the model group, the scratching head times of the control group, high- and low-dose groups of vinegar-stewed Cyperus rotundus, and high- and low-dose groups of raw Cyperus rotundus were all significantly decreased; when comparing raw and vinegar-stewed Cyperus rotundus at the same dose, the effect of vinegar-stewed Cyperus rotundus was better than that of raw Cyperus rotundus. The observation results are as follows, shown in Table 3.

[0065] Table 3 Comparison of the scratching head times of rats (n = 6, )

[0066]

[0067]

[0068] Note: * p < 0.05 compared with the blank group, with statistical significance; # p < 0.05 compared with the model group, with statistical significance.

[0069] 4.2.2 Effects of Vinegar-Stewed Cyperus rotundus on the Contents of 5-HT and CGRP in Rat Serum

[0070] Effect of Vinegar-braised Cyperus rotundus on the Contents of 5-HT and CGRP in Rat Serum: Compared with the blank group, the contents of 5-HT in the serum of rats in the model group, the low-dose vinegar Cyperus rotundus group, and the high- and low-dose raw Cyperus rotundus groups were significantly decreased; the contents of CGRP in the serum of rats in the model group, the high- and low-dose vinegar Cyperus rotundus groups, and the high- and low-dose raw Cyperus rotundus groups were significantly increased; compared with the model group, the 5-HT in the blank group, the control group, the high- and low-dose vinegar-braised Cyperus rotundus groups, and the high-dose raw Cyperus rotundus group was significantly increased; the contents of CGRP in the blank group, the control group, the high-dose vinegar-braised Cyperus rotundus group, and the high-dose raw Cyperus rotundus group were significantly decreased. The results are as follows, shown in Table 4.

[0071] Table 4 Levels of 5-HT and CGRP in the Serum of Rats in Each Group (n = 6, )

[0072]

[0073] Note: *Compared with the blank group, p < 0.05, with statistical significance; #Compared with the model group, p < 0.05, with statistical significance.

[0074] 4.3 Conclusion

[0075] The improvement of rat behavior indicates that vinegar-braised Cyperus rotundus has obvious preventive and therapeutic effects on migraine model rats. Vinegar-braised Cyperus rotundus can increase the content of 5-HT in the serum of migraine model rats and decrease the content of CGRP in the serum.

[0076] Typical Cases

[0077] Case 1: Song, male, 24 years old, with recurrent severe pain in the left temporal region, accompanied by nausea, vomiting, photophobia, and phonophobia. He was diagnosed with acute migraine. After using the processed product in Example 1 of the present invention for 10 days, the frequency of headache attacks was significantly reduced, the pain intensity was alleviated, the accompanying symptoms were significantly relieved, and the quality of daily life was improved.

[0078] Case 2: Wang, female, 35 years old, suffering from acute migraine for a long time. When the headache was severe, it was accompanied by dizziness and vomiting, affecting daily work. After using the processed product in Example 1 of the present invention for 7 days, the number of headache attacks decreased, the symptoms of dizziness and vomiting improved, and the working state recovered well.

Claims

1. A preparation method of a processed Cyperus rotundus product for treating migraine, characterized in that, It includes the following steps: Step 1: Add the purified Cyperus rotundus L. granules and pieces to rice vinegar, mix well, and then moisten them. Step 2: Then braise them in a casserole and dry them to obtain the processed Cyperus rotundus L. product for treating migraine.

2. The preparation method of the prepared Cyperus rotundus product for treating migraine according to claim 1, characterized in that, In Step 1, the mass of the rice vinegar used is 10%-30% of the mass of the purified Cyperus rotundus L. granules and pieces.

3. The preparation method of the processed Cyperus rotundus product for treating migraine according to claim 1, wherein In Step 1, the alcohol content of the rice vinegar is 3.5°-9°.

4. The preparation method of the prepared Cyperus rotundus product for treating migraine according to claim 1, wherein In Step 1, the moistening time is 4 hours.

5. The preparation method of the prepared product of Cyperus rotundus for treating migraine according to claim 1, characterized in that, In Step 2, the braising time in the casserole is 3 hours - 7 hours.

6. The preparation method of the processed Cyperus rotundus product for treating migraine according to claim 1, wherein, The application of the processed Cyperus rotundus L. product for treating migraine in the medicine for treating migraine.

7. The preparation method of the processed Cyperus rotundus product for treating migraine according to claim 1, wherein The processed Cyperus rotundus L. product for treating migraine can improve the scratching times of migraine rats and the levels of 5-HT and CGRP in the serum.

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