A Mongolian medicine for nourishing the liver and promoting hematopoiesis, its preparation method and application

Abstract

The present invention discloses a preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis, including: crushing tsaoko fruit, gypsum, viola yedoensis, momordica cochinchinensis, green tangerine peel, amomum kravanh, aquilaria sinensis, madder, costus root, dwarf lilyturf tuber, weeping forsythia, semen nelumbinis, cape jasmine fruit, nardostachys chinensis, etc. to obtain a prefabricated Mongolian medicine powder; adding cholesterol and lecithin to ethanol and stirring evenly to obtain a premixed solution; dissolving lactoferrin in PBS solution and stirring evenly, dropping the premixed solution into it under stirring, evaporating to remove ethanol, performing ultrasonic treatment for 1 - 2 h, then adjusting the temperature to 10 - 20 °C and standing for 1 - 4 h, and filtering to obtain lactoferrin liposomes; boiling honey and then changing to a gentle heat, continuing to boil until the foam shows a light red luster, adding konjac gum, propolis, and chitosan, adjusting the pH value of the system to 6.2 - 6.7, adding lactoferrin liposomes, and homogenizing to obtain a concentrated gel; fully mixing the prefabricated Mongolian medicine powder and the concentrated gel, making pills, wrapping the pills after sweating, and performing microwave sterilization to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

Description

Technical Field

[0001] The invention relates to the technical field of Mongolian medicines, and in particular to a liver-nourishing and hematopoietic Mongolian medicine and a preparation method and application thereof. Background Art

[0002] Aplastic anemia is one of the difficult and serious diseases of the blood system. It is a disease caused by a variety of causes, including bone marrow hematopoietic failure, decreased number or abnormal function of hematopoietic stem cells, and abnormal hematopoietic microenvironment, which leads to decreased red bone marrow capacity, erythropenia, neutropenia, thrombocytopenia and other blood cell reductions. At present, it is believed that the onset of aplastic anemia is related to abnormal immune mechanisms, defects in the hematopoietic microenvironment, and defects in the number and / or function of hematopoietic stem cells. The cause of the disease is unclear and may be related to infections such as hepatitis viruses and parvoviruses, chemical factors such as chloramphenicol antibiotics, sulfonamides and pesticides, or physical factors such as X-rays. Its pathogenesis has not yet been fully elucidated.

[0003] Western medicine is a common treatment for aplastic anemia, mainly iron injection or oral administration. However, its efficacy is general and has limitations. Clinical practice has found that after taking iron supplements, patients may reduce their medication compliance due to adverse reactions, which is not conducive to subsequent treatment.

[0004] Mongolian medicine belongs to the medicine of ethnic minorities in my country, with a long history and distinct ethnic characteristics. In the Mongolian medical theory system, the cause of aplastic anemia is the imbalance of three roots, namely, the imbalance of Bada Gan, He Yi and Xila. At present, Mongolian medicine has obvious therapeutic advantages, is more convenient to use, is less likely to cause adverse drug reactions, and is more acceptable to patients.

[0005] At present, Mongolian medicine has achieved good therapeutic effects, and its cure rate and effectiveness rate have been significantly improved. At the same time, Mongolian medicine is a pure green medicine, which has the functions of promoting blood circulation and removing blood stasis, improving the bone marrow hematopoietic microenvironment, is cheap and safe, and long-term use does not damage the patient's own immune function. The chance of death due to secondary infection is low, and it can eliminate or reduce the dosage and toxic side effects of hormones and immunosuppressants, which is worthy of clinical promotion. Summary of the invention

[0006] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a liver-nourishing and hematopoietic Mongolian medicine and a preparation method and application thereof.

[0007] A method for preparing a liver-nourishing and hematopoietic Mongolian medicine comprises the following steps:

[0008] S1. Grind tsaoko, gypsum, violet yedoensis, cochinchinensis, green peel, white cardamom, agarwood, madder, costusroot, ophiopogon japonicus, cinnamomum officinale, caesalpinia, gardenia, nard, terminalia chebula, jujube, santalinum, dragon's blood, saffron, musk, three fresh red, cardamom and cistanche to obtain prefabricated Mongolian medicine powder;

[0009] S2. Add cholesterol and lecithin to ethanol and stir evenly to obtain a premixed solution; dissolve lactoferrin in PBS solution and stir evenly. While stirring, dropwise add the premixed solution, evaporate to remove ethanol, perform ultrasonic treatment for 1 - 2 h, then adjust the temperature to 10 - 20 °C and let it stand for 1 - 4 h, and filter to obtain lactoferrin liposomes;

[0010] S3. Boil honey, then turn to low heat and continue to boil until the foam shows a light red luster. Add konjac gum, propolis, and chitosan, adjust the pH value of the system to 6.2 - 6.7, add lactoferrin liposomes, and homogenize to obtain a concentrated gel;

[0011] S4. Thoroughly mix the pre - prepared Mongolian medicine powder with the concentrated gel, make pills, wrap the pills after sweating, and perform microwave sterilization to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

[0012] Preferably, in S1, the mass ratio of tsaoko fruit, gypsum, viola philippica, momordica cochinchinensis, green tangerine peel, amomum kravanh, agarwood, madder, costus root, dwarf lilyturf tuber, weeping myrrh, semen nelumbinis, gardenia fruit, nardostachys jatamansi, terminalia chebula, fructus choerospondiatis, red sandalwood, dragon's blood, saffron, musk, three - fresh red, amomum kravanh, and desertliving cistanche is 45 - 55:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:8 - 12:350 - 450:8 - 12:4 - 6.

[0013] Preferably, in S1, after pulverization, it is sieved through a 120 - mesh sieve.

[0014] Preferably, in S2, the mass ratio of cholesterol, lecithin, and lactoferrin is 1 - 4:5 - 15:1 - 5.

[0015] Preferably, in S2, the ultrasonic treatment power is 200 - 400 W.

[0016] Preferably, in S2, the particle size of lactoferrin liposomes is 0.1 - 1.5 μm.

[0017] Preferably, in S3, the mass ratio of honey, konjac gum, propolis, chitosan, and lactoferrin liposomes is 10 - 20:1 - 5:1 - 5:1 - 2:10 - 25.

[0018] Preferably, in S4, the mass ratio of the pre - prepared Mongolian medicine powder to the concentrated gel is 90 - 110:20 - 30.

[0019] A Mongolian medicine for nourishing the liver and promoting hematopoiesis is prepared by the above - mentioned preparation method of the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

[0020] The application of the above - mentioned Mongolian medicine for nourishing the liver and promoting hematopoiesis in the preparation of drugs for treating aplastic anemia.

[0021] The technical effects of the present invention are as follows:

[0022] In the present invention, lactoferrin is encapsulated into phospholipid liposomes, which can not only form stable liposomes, but also use honey as raw material, cooperate with konjac gum and propolis, and can effectively enhance the stability of honey pills; the concentrated glue and the prefabricated Mongolian medicine powder are compounded into pills. Since the lactoferrin liposomes have strong negative charges, during the thorough mixing process, they can combine with chitosan through self-assembly, which can not only further enhance the overall stability of the prepared pills, but also make the content of reducing sugar in the honey pills ≤ 0.5 g / 100 g, and at the same time have excellent antibacterial effect, and the total number of colonies ≤ 10 cfu / g. After 90 days of accelerated experiment, all indicators of the product are stable, and the problems of sterilization and preservation of honey pills can be effectively solved.

[0023] The Mongolian medicine of the present invention is based on Mongolian medical theory, taking into account conditioning and treatment at the same time, and playing the role of preventing and treating diseases. It has better therapeutic effect on patients with aplastic anemia, can effectively regulate peripheral blood picture indexes, improve cellular immunity, and promote the recovery of bone marrow hematopoietic function, and has high safety; at the same time, the traditional Chinese medicine raw materials are combined with lactoferrin liposomes. Due to the excellent lipophilicity and mucosal permeability of lactoferrin liposomes, combined with chitosan, it can assist in entering cells, and then synergistically act with the Mongolian medicine composition. It can not only actively participate in and promote the improvement of cellular immunity, but also has good targeting, can greatly reduce the dosage of traditional Chinese medicine, and significantly improve the absorption efficiency. The applicant of the present invention found through experiments that: the compounding of lactoferrin liposomes and the Mongolian medicine composition in the present application, compared with only adding the Mongolian medicine composition, can not only regulate the disordered T cell subsets, improve cellular immunity, but also promote the recovery of bone marrow hematopoietic function, correct peripheral blood picture indexes, and at the same time will not increase adverse reactions, and the drug safety is high. Description of the Drawings

[0024] Figure 1 It is a comparison chart of white blood cells of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group in the immune-mediated aplastic anemia experiment.

[0025] Figure 2 It is a comparison chart of white blood cells of the model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group in the immune-mediated aplastic anemia experiment.

[0026] Figure 3 It is a comparison chart of red blood cells of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group in the immune-mediated aplastic anemia experiment.

[0027] Figure 4Comparison chart of hemoglobin of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the immune-mediated aplastic anemia test.

[0028] Figure 5 Comparison chart of platelets of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the immune-mediated aplastic anemia test.

[0029] Figure 6 Comparison chart of platelets of the model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the immune-mediated aplastic anemia test.

[0030] Figure 7 For the CD3 + T cell comparison chart of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the immune-mediated aplastic anemia test.

[0031] Figure 8 For the CD8 + / CD4 + Ratio comparison chart of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the immune-mediated aplastic anemia test.

[0032] Figure 9 Comparison chart of white blood cells of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the complex factor-induced aplastic anemia test.

[0033] Figure 10 Comparison chart of white blood cells of the model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the complex factor-induced aplastic anemia test.

[0034] Figure 11 Comparison chart of red blood cells of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the complex factor-induced aplastic anemia test.

[0035] Figure 12 Comparison chart of hemoglobin of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the complex factor-induced aplastic anemia test.

[0036] Figure 13 Comparison chart of platelets of the normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group in the complex factor-induced aplastic anemia test.

[0037] Figure 14 In the test of aplastic anemia induced by complex factors, the platelet comparison chart of the model group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3.

[0038] Figure 15 In the test of aplastic anemia induced by complex factors, the comparison chart of reticulocyte counts of the normal control group, the model group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3.

[0039] Figure 16 In the test of aplastic anemia induced by complex factors, CD3 + T cell comparison chart of the normal control group, the model group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3.

[0040] Figure 17 In the test of aplastic anemia induced by complex factors, CD8 + / CD4 + Ratio comparison chart of the normal control group, the model group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3. Detailed implementation mode

[0041] The present invention will be further explained below in conjunction with specific embodiments. Example 1

[0042] A preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis includes the following steps:

[0043] S1. Crush 45 g of tsaoko fruit, 25 g of gypsum, 25 g of viola yedoensis, 25 g of momordica cochinchinensis, 25 g of green tangerine peel, 25 g of amomum kravanh, 25 g of aquilaria sinensis, 25 g of madder, 25 g of Aucklandia lappa, 25 g of ophiopogon japonicus, 25 g of trogopterus dung, 25 g of semen nelumbinis, 25 g of gardenia jasminoides, 25 g of nardostachys jatamansi, 25 g of terminalia chebula, 25 g of fructus choerospondiatis, 25 g of red sandalwood, 25 g of dragon's blood, 25 g of crocus sativus, 8 g of moschus, 350 g of three fresh reds, 8 g of amomum kravanh, and 4 g of cistanche deserticola, and pass through a 120-mesh sieve to obtain a prefabricated Mongolian medicine powder;

[0044] S2. Add 1 g of cholesterol and 5 g of lecithin to 40 g of ethanol and stir evenly to obtain a premixed solution; dissolve 1 g of lactoferrin in 100 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. While stirring, dropwise add the premixed solution, evaporate to remove ethanol, perform ultrasonic treatment for 1 h with an ultrasonic treatment power of 200 W, then adjust the temperature to 10 °C and let it stand for 1 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm;

[0045] S3. Boil 10 g of honey until it boils, then turn to a low heat and continue boiling until the foam shows a light red luster. Add 1 g of konjac gum, 1 g of propolis, and 1 g of chitosan thereto, adjust the pH value of the system to 6.2 - 6.7, add 10 g of lactoferrin liposomes, and homogenize to obtain a concentrated gel.

[0046] S4. Thoroughly mix 90 g of prefabricated Mongolian medicine powder with 20 g of concentrated gel, make pills, wrap the pills after sweating, and sterilize them by microwave at 1000 MHz for 1 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis. Example 2

[0047] A preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0048] S1. Crush 55 g of tsaoko fruit, 35 g of gypsum, 35 g of viola yedoensis, 35 g of momordica cochinchinensis, 35 g of immature tangerine peel, 35 g of amomum kravanh, 35 g of aquilaria sinensis, 35 g of rubia cordifolia, 35 g of costus root, 35 g of ophiopogon japonicus, 35 g of dung beetle, 35 g of semen nelumbinis, 35 g of gardenia jasminoides, 35 g of nardostachys jatamansi, 35 g of terminalia chebula, 35 g of choerospondias axillaris, 35 g of red sandalwood, 35 g of dragon's blood, 35 g of crocus sativus, 12 g of moschus, 450 g of three fresh reds, 12 g of cardamom, and 6 g of cistanche deserticola, and pass through a 120 - mesh sieve to obtain prefabricated Mongolian medicine powder.

[0049] S2. Add 4 g of cholesterol and 15 g of lecithin to 60 g of ethanol and stir evenly to obtain a premixed solution; dissolve 5 g of lactoferrin in 200 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. While stirring, drop the premixed solution into it, evaporate to remove ethanol, perform ultrasonic treatment for 2 h with an ultrasonic treatment power of 400 W, then adjust the temperature to 20 °C and let it stand for 4 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm.

[0050] S3. Boil 20 g of honey until it boils, then turn to a low heat and continue boiling until the foam shows a light red luster. Add 5 g of konjac gum, 5 g of propolis, and 2 g of chitosan thereto, adjust the pH value of the system to 6.2 - 6.7, add 25 g of lactoferrin liposomes, and homogenize to obtain a concentrated gel.

[0051] S4. Thoroughly mix 110 g of prefabricated Mongolian medicine powder with 30 g of concentrated gel, make pills, wrap the pills after sweating, and sterilize them by microwave at 1000 MHz for 2 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis. Example 3

[0052] A preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0053] S1. Crush 48 g of tsaoko fruit, 32 g of gypsum, 28 g of viola yedoensis, 32 g of cochinchina momordica seed, 28 g of immature tangerine peel, 32 g of round cardamom, 28 g of aquilaria sinensis, 32 g of madder, 28 g of costus root, 32 g of dwarf lilyturf tuber, 28 g of dung beetle, 32 g of semen nelumbinis, 28 g of cape jasmine fruit, 32 g of Chinese spikenard, 28 g of myrobalan fruit, 32 g of choerospondias axillaris, 28 g of red sandalwood, 32 g of dragon's blood, 28 g of saffron, 11 g of moschus, 380 g of three fresh reds, 11 g of cardamom and 4.5 g of cistanche salsa, and pass through a 120-mesh sieve to obtain the prefabricated Mongolian medicine powder;

[0054] S2. Add 3 g of cholesterol and 8 g of lecithin to 55 g of ethanol and stir evenly to obtain a premixed solution; dissolve 2 g of lactoferrin in 180 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. While stirring, add the premixed solution dropwise, evaporate to remove ethanol, perform ultrasonic treatment for 80 min with an ultrasonic treatment power of 350 W, then adjust the temperature to 12 °C and let it stand for 3 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm;

[0055] S3. Boil 13 g of honey, then change to a slow fire and continue to boil until the foam shows a light red luster. Add 4 g of konjac gum, 2 g of propolis, and 1.7 g of chitosan, adjust the pH value of the system to 6.2 - 6.7, add 15 g of lactoferrin liposomes, and homogenize to obtain a concentrated gel;

[0056] S4. Thoroughly mix 105 g of the prefabricated Mongolian medicine powder with 22 g of the concentrated gel, make pills, wrap the pills after sweating, and sterilize with 1000 MHz microwave for 1.5 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis. Example 4

[0057] A method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0058] S1. Crush 52 g of tsaoko fruit, 28 g of gypsum, 32 g of viola yedoensis, 28 g of cochinchina momordica seed, 32 g of immature tangerine peel, 28 g of round cardamom, 32 g of aquilaria sinensis, 28 g of madder, 32 g of costus root, 28 g of dwarf lilyturf tuber, 32 g of dung beetle, 28 g of semen nelumbinis, 32 g of cape jasmine fruit, 28 g of Chinese spikenard, 32 g of myrobalan fruit, 28 g of choerospondias axillaris, 32 g of red sandalwood, 28 g of dragon's blood, 32 g of saffron, 9 g of moschus, 420 g of three fresh reds, 9 g of cardamom and 5.5 g of cistanche salsa, and pass through a 120-mesh sieve to obtain the prefabricated Mongolian medicine powder;

[0059] S2. Add 2 g of cholesterol and 12 g of lecithin to 45 g of ethanol and stir evenly to obtain a premixed solution; dissolve 4 g of lactoferrin in 120 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. While stirring, add the premixed solution dropwise, evaporate to remove ethanol, perform ultrasonic treatment for 100 min with an ultrasonic treatment power of 250 W, then adjust the temperature to 18 °C and let it stand for 2 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm;

[0060] S3. Boil 17 g of honey until it boils, then turn to a gentle heat and continue boiling until the foam shows a light red luster. Add 2 g of konjac gum, 4 g of propolis, and 1.3 g of chitosan to it, adjust the pH value of the system to 6.2 - 6.7, add 20 g of lactoferrin liposomes, and homogenize to obtain a concentrated gel.

[0061] S4. Thoroughly mix 95 g of prefabricated Mongolian medicine powder with 28 g of concentrated gel, make pills, wrap the pills after sweating, and sterilize them with 1000 MHz microwave for 1.5 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis. Example 5

[0062] A method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0063] S1. Crush 50 g of tsaoko fruit, 30 g of gypsum, 30 g of viola yedoensis, 30 g of momordica cochinchinensis, 30 g of immature tangerine peel, 30 g of amomum kravanh, 30 g of agarwood, 30 g of madder, 30 g of costus root, 30 g of dwarf lilyturf tuber, 30 g of trogopterus dung, 30 g of semen nelumbinis, 30 g of gardenia fruit, 30 g of nardostachys jatamansi, 30 g of myrobalan fruit, 30 g of wild jujube, 30 g of red sandalwood, 30 g of dragon's blood, 30 g of saffron, 10 g of musk, 400 g of fresh three - red, 10 g of cardamom, and 5 g of cistanche deserticola, and pass through a 120 - mesh sieve to obtain prefabricated Mongolian medicine powder.

[0064] S2. Add 2.5 g of cholesterol and 10 g of lecithin to 50 g of ethanol and stir evenly to obtain a premixed solution; dissolve 3 g of lactoferrin in 150 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. Drop the premixed solution into it under stirring, evaporate to remove ethanol, perform ultrasonic treatment for 90 min with an ultrasonic treatment power of 300 W, then adjust the temperature to 15 °C and let it stand for 2.5 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm.

[0065] S3. Boil 15 g of honey until it boils, then turn to a gentle heat and continue boiling until the foam shows a light red luster. Add 3 g of konjac gum, 3 g of propolis, and 1.5 g of chitosan to it, adjust the pH value of the system to 6.2 - 6.7, add 18 g of lactoferrin liposomes, and homogenize to obtain a concentrated gel.

[0066] S4. Thoroughly mix 100 g of prefabricated Mongolian medicine powder with 25 g of concentrated gel, make pills, wrap the pills after sweating, and sterilize them with 1000 MHz microwave for 1.5 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

[0067] Control Example 1

[0068] Use Xuekang Capsules (manufacturer: Jiangxi Tianshikang Traditional Chinese Medicine Co., Ltd.).

[0069] Control Example 2

[0070] A preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0071] S1. Crush 50 g of tsaoko fruit, 30 g of gypsum, 30 g of viola yedoensis, 30 g of cochinchina momordica seed, 30 g of green tangerine peel, 30 g of amomum kravanh, 30 g of aquilaria sinensis, 30 g of madder, 30 g of costus root, 30 g of dwarf lilyturf tuber, 30 g of oriental bittersweet fruit, 30 g of stone lotus seed, 30 g of cape jasmine fruit, 30 g of nardostachys chinensis, 30 g of myrobalan fruit, 30 g of choerospondias axillaris, 30 g of red sandalwood, 30 g of dragon's blood, 30 g of saffron, 10 g of musk, 400 g of three fresh reds, 10 g of cardamom and 5 g of cistanche deserticola, and pass through a 120-mesh sieve to obtain the prefabricated Mongolian medicine powder;

[0072] S3. Boil 15 g of honey, then change to a gentle heat and continue boiling until the foam shows a light red luster. Add 3 g of konjac gum, 3 g of propolis and 1.5 g of chitosan thereto, adjust the pH value of the system to 6.2 - 6.7, add 18 g of lactoferrin, and homogenize to obtain the concentrated glue;

[0073] S4. Thoroughly mix 100 g of the prefabricated Mongolian medicine powder with 25 g of the concentrated glue, make pills, wrap the pills after sweating, and sterilize them by 1000 MHz microwave for 1.5 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

[0074] Comparative Example 3

[0075] A preparation method of Mongolian medicine for nourishing the liver and promoting hematopoiesis, comprising the following steps:

[0076] S1. Crush 50 g of tsaoko fruit, 30 g of gypsum, 30 g of viola yedoensis, 30 g of cochinchina momordica seed, 30 g of green tangerine peel, 30 g of amomum kravanh, 30 g of aquilaria sinensis, 30 g of madder, 30 g of costus root, 30 g of dwarf lilyturf tuber, 30 g of oriental bittersweet fruit, 30 g of stone lotus seed, 30 g of cape jasmine fruit, 30 g of nardostachys chinensis, 30 g of myrobalan fruit, 30 g of choerospondias axillaris, 30 g of red sandalwood, 30 g of dragon's blood, 30 g of saffron, 10 g of musk and 400 g of three fresh reds, and pass through a 120-mesh sieve to obtain the prefabricated Mongolian medicine powder;

[0077] S2. Add 2.5 g of cholesterol and 10 g of lecithin to 50 g of ethanol and stir evenly to obtain a premixed solution; dissolve 3 g of lactoferrin in 150 g of PBS solution with a pH of 6.2 - 7.1 and stir evenly. Dropwise add the premixed solution thereto under stirring, evaporate to remove ethanol, perform ultrasonic treatment for 90 min with an ultrasonic treatment power of 300 W, then adjust the temperature to 15 °C and let it stand for 2.5 h, and filter with a filter membrane to obtain lactoferrin liposomes with a particle size of 0.1 - 1.5 μm;

[0078] S3. Boil 15 g of honey, then change to a gentle heat and continue boiling until the foam shows a light red luster. Add 3 g of konjac gum, 3 g of propolis and 1.5 g of chitosan thereto, adjust the pH value of the system to 6.2 - 6.7, add 18 g of lactoferrin liposomes, and homogenize to obtain the concentrated glue;

[0079] S4. Mix 100 g of the prepared Mongolian medicine powder and 25 g of the concentrated glue evenly, make pills, wrap the pills after sweating, and sterilize them with 1000 MHz microwave for 1.5 min to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

[0080] Using mice as the test subjects, a comparative test on the treatment of aplastic anemia was carried out with the Mongolian medicine for nourishing the liver and promoting hematopoiesis obtained in Example 5 and Comparative Examples 2 - 3 and the Xuekang Capsule used in Comparative Example 1.

[0081] (1) Immune - mediated aplastic anemia

[0082] Take 8 8 - week - old DBA / 2 mice (weight 18 - 20 g, male, SPF grade), collect blood by eye - ball enucleation and sacrifice them, remove the thymus and lymph nodes in the axilla, groin, submandibular, mesenteric and other places, and make the cell activity reach more than 95%, and prepare a single - cell suspension with a cell number of 5×10 6 / ml. Mix the thymus cells and lymph node cells in a ratio of 1:2 to prepare a mixed single - cell suspension for standby.

[0083] Take 72 BALB / c mice (6 - 8 weeks old, weight 16 - 18 g, female, SPF grade), randomly divide them into 6 groups: normal control group, model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group, with 12 mice in each group. Except for the normal control group, the other 5 groups of mice were irradiated with 5.0 Gy of X - rays from a linear accelerator throughout the body, and within 4 h, 2 mL of the mixed suspension of thymus and lymph node cells from DBA / 2 mice with a cell number of 1×10 6 was injected into the tail vein to establish an immune - mediated aplastic anemia model.

[0084] Taking pancytopenia; bone marrow pathology showing hypoplasia or hypofunction in multiple parts of the bone marrow, and an increase in the proportion of non - hematopoietic cells; bone marrow nucleated cell count showing a significant decrease in the number of bone marrow nucleated cells and other changes as the successful indicators of model establishment.

[0085] While establishing the immune - mediated aplastic anemia model, the normal control group and the model group were given 20 mL / kg of distilled water by gavage; the Comparative Example 1 group was given Xuekang Capsule suspension by gavage daily at a dose of 1.5 g / kg; the Example 5 group, Comparative Example 2 group and Comparative Example 3 group were respectively given the Mongolian medicine for nourishing the liver and promoting hematopoiesis suspension obtained in Example 5 and Comparative Examples 2 - 3 by gavage daily at a dose of 1.5 g / kg. The administration lasted for 15 days in total.

[0086] One hour after the last administration, collect blood by eye - ball enucleation, and detect white blood cells (WBC), red blood cells (RBC), hemoglobin (HGB), platelets (PLT), and T - cell subsets (CD3, CD4, CD8).

[0087] Among them, the data of white blood cells, red blood cells, hemoglobin, and platelets are respectively as Figure 1 、Figure 3 , Figure 4 and Figure 5 as shown. Since the white blood cell and platelet data of the normal control group far exceeded those of other groups, the applicant only redrew the corresponding data of the model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group, as shown in Figure 2 and Figure 6 .

[0088] It can be seen from Figures 1 - 6 that there were significant differences in white blood cells, red blood cells, hemoglobin, and platelets between the model group and the normal control group. Compared with the normal control group, the white blood cells, red blood cells, hemoglobin, and platelets in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group were significantly reduced; while compared with the model group, the white blood cells, red blood cells, hemoglobin, and platelets in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group were significantly increased.

[0089] At the same time, when comparing the Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group with each other, it was found that the contents of white blood cells, red blood cells, hemoglobin, and platelets in the Example 5 group were the highest.

[0090] The contents of T cell subsets (CD3 + ) and the ratio of CD8 + / CD4 + were as shown in Figure 7 and Figure 8 respectively. There were significant differences in the content of CD3 + and the ratio of CD8 + / CD4 + between the model group and the normal control group. Compared with the model group, the content of CD3 + in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group was significantly increased, while the ratio of CD8 + / CD4 + was significantly reduced. When comparing the Example 5 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group with each other, it was found that the content of CD3 + in the Example 5 group was the highest, and the ratio of CD8 + / CD4 + was the lowest.

[0091] The mouse immune-mediated aplastic anemia model is established by irradiating mice with sublethal doses of radiation and then injecting a mixture of thymus and lymph node cells from mice with incompatible histocompatibility antigens. Due to damage to the body's immune system, these cells survive in the recipient mice and further act on the bone marrow hematopoietic tissue through a certain pathway, resulting in bone marrow hematopoietic failure. Based on the above-mentioned peripheral blood picture and peripheral blood T lymphocyte subset indicators, it shows that the Yanggan Zaoxue Mongolian medicine obtained in Example 5 and Comparative Examples 2-3 and the Xuekang Capsule used in Comparative Example 1 have therapeutic effects on immune-mediated aplastic anemia, and the Yanggan Zaoxue Mongolian medicine obtained in this application has the best therapeutic effect.

[0092] (2)Compound factor-induced aplastic anemia

[0093] Take 96 Wistar rats (weighing 180-220 g, half male and half female, SPF grade), and randomly divide them into 6 groups, namely the normal control group, the model group, the group of Example 5, the group of Comparative Example 1, the group of Comparative Example 2, and the group of Comparative Example 3, with 16 rats in each group.

[0094] On the first day of modeling, except for the normal control group, the other 5 groups of rats were fixed with a plastic fixator and irradiated with a linear accelerator at a total dose rate of 240 cGy and SSD = 100 cm throughout the body. Starting from the 4th day, cyclophosphamide 35 mg / kg and chloramphenicol 43.75 mg / kg were intraperitoneally injected every other day for a total of 3 times to establish a compound factor-induced aplastic anemia model.

[0095] Taking the reduction of the three-line cells in peripheral blood; the reduction of reticulocytes; and the bone marrow biopsy showing poor or decreased bone marrow hyperplasia (hematopoietic cell volume < 40%) and an increase in non-hematopoietic cells (adipocytes) as the success indicators of modeling.

[0096] At the same time of modeling, the normal control group and the model group were given 20 ml / kg of distilled water by gavage; the group of Comparative Example 1 was given Xuekang Capsule suspension by gavage daily at a dose of 1.5 g / kg; the group of Example 5, the group of Comparative Example 2, and the group of Comparative Example 3 were respectively given the Yanggan Zaoxue Mongolian medicine suspension obtained in Example 5 and Comparative Examples 2-3 by gavage daily at a dose of 1.5 g / kg. The administration lasted for 15 days in total.

[0097] One hour after the last administration, blood was collected from the abdominal aorta to detect the peripheral blood WBC, RBC, HGB, PLT, reticulocyte count (RET), and T cell subsets (CD3, CD4, CD8) of the rats.

[0098] Among them, the data of white blood cells, red blood cells, hemoglobin, platelets, and reticulocyte count are respectively as Figure 9 、 Figure 11 、 Figure 12 、 Figure 13 and Figure 15As shown. Since the white blood cell and platelet data of the normal control group far exceeded those of other groups, the applicant only redrew the corresponding data of the model group, Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group, as Figure 10 and Figure 14 shown.

[0099] It can be seen from Figures 9 - 15 that there were significant differences in the white blood cells, red blood cells, hemoglobin, platelets and reticulocyte counts between the model group and the normal control group. Compared with the normal control group, the white blood cells, red blood cells, hemoglobin and platelets in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group were significantly reduced; while compared with the model group, the white blood cells, red blood cells, hemoglobin, platelets and reticulocyte counts in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group were significantly increased.

[0100] At the same time, when comparing the Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group with each other, it was found that the contents of white blood cells, red blood cells, hemoglobin and platelets and the reticulocyte count in the Example 5 group were the highest.

[0101] The content of T cell subsets (CD3 + ) and the ratio of CD8 + / CD4 + are shown in Figure 16 and Figure 17 respectively. There were significant differences in the content of CD3 + and the ratio of CD8 + / CD4 + between the model group and the normal control group. Compared with the model group, the content of CD3 + in the Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group was significantly increased, while the ratio of CD8 + / CD4 + was significantly reduced. When comparing the Example 5 group, Comparative Example 1 group, Comparative Example 2 group and Comparative Example 3 group with each other, it was found that the content of CD3 + in the Example 5 group was the highest, and the ratio of CD8 + / CD4 + was the lowest.

[0102] The combined method is mainly caused by the combined action of main radiation, cyclophosphamide and chloramphenicol to inhibit DNA synthesis and act on the hematopoietic microenvironment or hematopoietic stem cells, thereby affecting the generation and maturation of blood cells and leading to the failure of bone marrow hematopoietic function. Based on the above peripheral blood picture and peripheral blood T lymphocyte subset indicators, it shows that the liver-nourishing and hematopoiesis-promoting Mongolian medicine obtained in Example 5 and Comparative Examples 2-3 and the Xuekang Capsule used in Comparative Example 1 have a therapeutic effect on aplastic anemia induced by combined factors, and the liver-nourishing and hematopoiesis-promoting Mongolian medicine obtained in this application has the best therapeutic effect.

[0103] Since T lymphocytes are the main effector cells in cellular immunity, research has found that in patients with aplastic anemia, there are not only abnormal numbers of T cells, but also obvious changes in the distribution, phenotype and function of T cell subsets. The above experimental results show that in both models, the number of CD3 + T cells increased in the fifth embodiment group, and the CD4 + / CD8 + ratio returned to the normal level.

[0104] The applicant of the present application speculates that the liver-nourishing and hematopoiesis-promoting Mongolian medicine obtained in the present application increases the relatively disadvantaged CD4 + T cells, inhibits the overly differentiated and activated CD8 + T cells, and restores the CD4 + / CD8 + T cell ratio to the normal level, thereby regulating the disorder of T cell subsets.

[0105] That is, the liver-nourishing and hematopoiesis-promoting Mongolian medicine obtained in the present application may promote the recovery of bone marrow hematopoiesis function and improve cellular immunity by regulating the disordered T cell subsets and reducing the targeting effect of Ts (CD8 + ) on bone marrow hematopoietic cells.

[0106] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, makes equivalent substitutions or changes, and should be covered by the protection scope of the present invention.

Claims

1. A method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis, characterized in that, It includes the following steps: S1. Grind tsaoko fruit, gypsum, viola philippica, momordica cochinchinensis, green tangerine peel, amomum kravanh, aquilaria sinensis, madder, costus root, dwarf lilyturf tuber, weeping forsythia fruit, cape jasmine fruit, nardostachys jatamansi, myrobalan fruit, mongolian snakegourd fruit, saffron crocus, moschus, three fresh reds, amomum kravanh, and desertliving cistanche to obtain a prefabricated Mongolian medicine powder; Among them, the mass ratio of tsaoko fruit, gypsum, viola philippica, momordica cochinchinensis, green tangerine peel, amomum kravanh, aquilaria sinensis, madder, costus root, dwarf lilyturf tuber, weeping forsythia fruit, cape jasmine fruit, nardostachys jatamansi, myrobalan fruit, mongolian snakegourd fruit, saffron crocus, moschus, three fresh reds, amomum kravanh, and desertliving cistanche is 45 - 55:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:25 - 35:8 - 12:350 - 450:8 - 12:4 - 6; S2. Add cholesterol and lecithin to ethanol and stir evenly to obtain a premixed solution; dissolve lactoferrin in PBS solution and stir evenly. While stirring, drop the premixed solution into it, evaporate to remove ethanol, perform ultrasonic treatment for 1 - 2 h, then adjust the temperature to 10 - 20 °C and let it stand for 1 - 4 h, and filter to obtain lactoferrin liposomes; S3. After boiling honey, change to a slow fire and continue to boil until the foam shows a light red luster. Add konjac gum, propolis, and chitosan to it, adjust the pH value of the system to 6.2 - 6.7, add lactoferrin liposomes, and homogenize to obtain a concentrated gel; S4. Thoroughly mix the prefabricated Mongolian medicine powder and the concentrated gel, make pills, wrap the pills after sweating, and perform microwave sterilization to obtain the Mongolian medicine for nourishing the liver and promoting hematopoiesis.

2. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S1, after grinding, it is sieved through a 120 - mesh sieve.

3. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S2, the mass ratio of cholesterol, lecithin, and lactoferrin is 1 - 4:5 - 15:1 - 5.

4. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S2, the ultrasonic treatment power is 200 - 400 W.

5. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S2, the particle size of the lactoferrin liposomes is 0.1 - 1.5 μm.

6. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S3, the mass ratio of honey, konjac gum, propolis, chitosan, and lactoferrin liposomes is 10 - 20:1 - 5:1 - 5:1 - 2:10 - 25.

7. The method for preparing Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 1, characterized in that, In S4, the mass ratio of the prefabricated Mongolian medicine powder and the concentrated gel is 90 - 110:20 - 30.

8. A Mongolian medicine for nourishing the liver and promoting hematopoiesis, characterized in that, Prepared by using the preparation method of the Mongolian medicine for nourishing the liver and promoting hematopoiesis according to any one of claims 1 - 7.

9. An application of the Mongolian medicine for nourishing the liver and promoting hematopoiesis according to claim 8 in the preparation of a medicine for treating aplastic anemia.

Images