CN113134040B

CN113134040B - Qi-tonifying blood-controlling traditional Chinese medicine composition for treating ITP and preparation method and application thereof

Info

Publication number: CN113134040B
Application number: CN202110616136.5A
Authority: CN
Inventors: 黄伟; 罗雅琴
Original assignee: Shandong Academy of Chinese Medicine
Current assignee: Shandong Academy of Chinese Medicine
Priority date: 2021-06-02
Filing date: 2021-06-02
Publication date: 2022-08-12
Anticipated expiration: 2041-06-02
Also published as: CN113134040A

Abstract

The invention belongs to the field of traditional Chinese medicines, and discloses a traditional Chinese medicine composition for benefiting qi and controlling blood circulation for treating ITP, which is prepared from the following raw materials in parts by weight: 25-35 parts of astragalus membranaceus, 20-30 parts of radix rehmanniae recen, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 25-35 parts of buffalo horn, 10-15 parts of red peony root, 6-12 parts of cortex moutan, 25-35 parts of hairyvein agrimonia herb and bud, 10-20 parts of madder, 10-20 parts of eclipta, 10-15 parts of selaginella tamariscina, 1-5 parts of pseudo-ginseng powder, 6-12 parts of glabrous sarcandra herb and 3-9 parts of liquorice. The qi-tonifying and blood-controlling formula with astragalus and astragalus membranaceus has definite clinical curative effect, obviously improves the symptoms of ITP patients, and increases the PLT count. Can increase the level of bone marrow megakaryocyte and TPO by adjusting the secretion imbalance of proinflammatory factors and inflammation-inhibiting factors of an ITP model mouse, thereby playing a role in treating ITP.

Description

Qi-tonifying blood-controlling traditional Chinese medicine composition for treating ITP and preparation method and application thereof

Technical Field

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a qi-tonifying blood-controlling prescription for treating ITP and an anti-inflammatory mechanism.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Immune Thrombocytopenia (ITP), a clinically common hemorrhagic disease, is characterized by a reduced number of platelets in the peripheral blood. Mild patients may have no obvious bleeding symptoms, severe patients may have skin mucosa bleeding, menorrhagia, even visceral and intracranial bleeding, and endanger life. The quality of life of ITP patients is seriously reduced, even lower than that of cancer patients, because of various factors such as increased bleeding risk, unpredictable disease outcome, fear to diseases, long-time treatment, reduced social activities, influence on work and the like. Currently, the first line treatment for ITP mainly involves: glucocorticoids, intravenous immunoglobulins and anti-D immunoglobulins; the second line treatment method comprises: splenectomy, thrombopoietin receptor agonist, rituximab, cyclosporine, immunosuppressant and the like. A significant proportion of patients do not respond well to first-and second-line treatment, and persistent repetitive treatment becomes refractory ITP, severely increasing patient burden. Clinical practice proves that the traditional Chinese medicine is used as a main or auxiliary treatment, and has obvious advantages in the aspects of preventing and treating bleeding of ITP patients, stabilizing the number of peripheral blood platelets, improving clinical symptoms and the like, for example: clinical research on spleen-invigorating, qi-tonifying and blood-controlling prescription for treating spleen-qi deficiency type immune thrombocytopenia requires deep research on the effect mechanism of traditional Chinese medicine treatment to better enhance the curative effect.

Disclosure of Invention

In order to overcome the problems, the invention provides a qi-tonifying and blood-controlling traditional Chinese medicine composition for treating ITP and a preparation method and application thereof, and the clinical treatment effect on ITP is effectively improved by researching the treatment effect and the anti-inflammatory mechanism of a qi-tonifying and blood-controlling prescription of astragalus mongholicus on an ITP model mouse.

In order to achieve the technical purpose, the invention adopts the following technical scheme:

the invention provides a traditional Chinese medicine composition for benefiting qi and controlling blood circulation for treating ITP, which comprises the following raw materials in parts by weight: 25-35 parts of astragalus membranaceus, 20-30 parts of radix rehmanniae recen, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 25-35 parts of buffalo horn, 10-15 parts of red peony root, 6-12 parts of cortex moutan, 25-35 parts of hairyvein agrimonia herb and bud, 10-20 parts of madder, 10-20 parts of eclipta, 10-15 parts of selaginella tamariscina, 1-5 parts of pseudo-ginseng powder, 6-12 parts of glabrous sarcandra herb and 3-9 parts of liquorice.

The existing research shows that: the qi-tonifying and blood-controlling formula which is mainly composed of astragalus, poria cocos, bighead atractylodes rhizome and the like can effectively improve clinical symptoms of patients and improve bleeding conditions, but the treatment mechanism is not clear, and the efficacy is to be enhanced. Therefore, the invention provides a clinical empirical formula (Qihuang qi-tonifying blood-controlling formula) for treating ITP according to the traditional Chinese medicine syndrome differentiation treatment theory and the summary of years of clinical experience, the clinical empirical formula mainly comprises 14 traditional Chinese medicines such as astragalus, radix rehmanniae, poria cocos, bighead atractylodes rhizome, buffalo horn and the like, the clinical curative effect is definite, the symptoms of ITP patients are obviously improved, and the PLT count is increased. Meanwhile, the invention also researches the treatment effect of the qi-tonifying and blood-taking formula of astragalus mongholicus on the ITP model mouse by establishing an ITP mouse model and continuously performing gastric lavage and drug administration for 14 days, discusses the anti-inflammatory mechanism of the blood-taking formula of astragalus mongholicus by respectively detecting the level changes of proinflammatory factors and anti-inflammatory factors in serum, and provides scientific basis and support for the clinical application of the blood-taking formula of astragalus mongholicus.

In a second aspect of the invention, a pharmaceutical preparation for treating ITP is provided, which is prepared from any one of the above-mentioned Chinese medicinal compositions and pharmaceutically acceptable excipients.

In a third aspect of the present invention, a preparation method of a qi-tonifying blood-nourishing traditional Chinese medicine composition for treating ITP is provided, which comprises:

mixing any of the above Chinese medicinal materials, extracting with water, collecting filtrate, concentrating, and drying.

The fourth aspect of the present invention provides an application of any one of the above-mentioned Chinese medicinal compositions in regulating the imbalance of secretion of proinflammatory factors and anti-inflammatory factors and increasing the level of bone marrow metaplastic megakaryocytes and TPO.

The invention has the beneficial effects that:

(1) the qi-tonifying and blood-controlling Qihuang prescription for treating ITP has definite clinical curative effect, obviously improves the symptoms of ITP patients and increases PLT (total viable cell count).

(2) The qi-tonifying and blood-taking formula of astragalus mongholicus can increase the levels of bone marrow megakaryocytes and TPO (platelet-derived macrophage) by regulating the secretion imbalance of proinflammatory factors and inflammation-inhibiting factors of an ITP model mouse, and plays a role in treating ITP.

(3) The preparation method is simple, low in cost, high in practicability and easy for large-scale production.

Detailed Description

It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

The invention provides a traditional Chinese medicine composition for benefiting qi and controlling blood circulation for treating ITP, which is prepared from the following raw materials in parts by weight: 25-35 parts of astragalus membranaceus, 20-30 parts of radix rehmanniae recen, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 25-35 parts of buffalo horn, 10-15 parts of red peony root, 6-12 parts of cortex moutan, 25-35 parts of hairyvein agrimonia herb and bud, 10-20 parts of madder, 10-20 parts of eclipta, 10-15 parts of selaginella tamariscina, 1-5 parts of pseudo-ginseng powder, 6-12 parts of glabrous sarcandra herb and 3-9 parts of liquorice.

Meanwhile, the therapeutic effect and the anti-inflammatory mechanism of the qi-tonifying and blood-controlling formula of astragalus membranaceus and astragalus membranaceus on Immune Thrombocytopenia (ITP) model mice are researched.

The specific method comprises the following steps: 50 BALB/C mice were randomly divided into 5 groups, namely a normal group, a model group, a Qihuang qi-tonifying and blood-engorging high and low dose group and a prednisone group, and 10 mice in each group. Establishing an ITP mouse model by adopting a guinea pig anti-mouse platelet serum (APS) intraperitoneal injection method; on day 5 after APS injection, each fraction was administered by intragastric administration for 14 days. Observing the general condition of each group of mice; detecting changes in peripheral platelet count (PLT) and hemoglobin (Hb) concentration; the Elisa method detects the levels of serum Thrombopoietin (TPO), proinflammatory factor interleukin-2 (IL-2), IL-6, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and inflammation suppressor factors IL-4, IL-10 and transforming growth factor-beta 1 (TGF-beta 1); separating spleen and thymus gland tissues, weighing, and calculating organ index; taking the sternum as a bone marrow smear, and carrying out bone marrow megakaryocyte classification under a microscope.

The research result shows that: compared with a normal group, the peripheral blood PLT count, Hb and TPO level of the model group mice are remarkably reduced (P <0.05 or P <0.01), spleen and thymus index are remarkably increased (P <0.01), serum IL-2, IL-6, TNF-alpha and IFN-gamma levels are remarkably increased (P <0.05 or P <0.01), IL-4, IL-10 and TGF-beta 1 levels are remarkably reduced (P <0.01), and the number of megakaryocytes produced from bone marrow is remarkably reduced (P < 0.01); compared with the model group, the PLT counts and TPO levels of the astragalus mongholicus blood-tonifying formula high and low dose groups and the prednisone group are obviously increased (P is less than 0.01), the spleen index and the thymus index are obviously reduced (P is less than 0.05 or P is less than 0.01), the levels of serum IL-2, IL-6, TNF-alpha and IFN-gamma are obviously reduced (P is less than 0.05 or P is less than 0.01), the levels of IL-4, IL-10 and TGF-beta 1 are obviously increased (P is less than 0.05 or P is less than 0.01), the number of bone marrow plate-producing megakaryocytes is obviously increased (P is less than 0.01), and the Hb level of the high dose group is obviously increased (P is less than 0.05) only; compared with the low dose group, the PLT counts of the high dose group and the prednisone group are obviously increased (P is less than 0.05), the TNF-alpha level is obviously reduced (P is less than 0.05 or P is less than 0.01), the IL-4 and TGF-beta 1 levels are obviously increased (P is less than 0.01), the IL-6 level of only the prednisone group is obviously reduced (P is less than 0.05), the IL-10 level of only the high dose group is obviously increased (P is less than 0.05), and the rest indexes have no statistical difference (P is more than 0.05).

The present invention is described in further detail below with reference to specific examples, which are intended to be illustrative of the invention and not limiting.

Example 1:

1 materials

1.1 Experimental animals

SPF-grade BALB/C mice, the body mass is 18-20 g, the sex is half each, Beijing vitamin Tonglihua experimental animal breeding Limited company, the animal license number: SCXK (Kyoto) 2016-. Conventional feeding in animal rooms of Shandong Chinese medicinal university, wherein the feeding conditions are as follows: the illumination rhythm is 12L: 12D (6: 00-18: 00); room temperature is 20-25 ℃; relative humidity (50 +/-5)%.

1.2 Experimental drugs

The qi-tonifying and blood-nourishing formula is an empirical formula for clinical treatment of ITP in the department, and comprises 30g of astragalus, 24g of radix rehmanniae recen, 15g of poria cocos, 15g of bighead atractylodes rhizome, 30g of buffalo horn, 12g of red peony root, 9g of cortex moutan, 30g of hairyvein agrimony, 15g of madder, 15g of eclipta, 12g of selaginella tamariscina, 3g of pseudo-ginseng powder, 9g of glabrous sarcandra herb and 6g of liquorice, wherein the daily prescription amount is 225 g. The above medicinal materials are provided by Beijing Tongrentang pharmaceutical industry GmbH, and are certified by Bin researchers at traditional Chinese medicine resource research institute of Shandong province. The 14 medicinal materials of the whole formula are added with 10 times of water for extraction for 2 times, each time lasts for 1 hour, the filtrates are combined, concentrated, dried in vacuum, crushed into fine powder and prepared into liquid medicine with required concentration by using distilled water before use. Prednisone acetate tablets: the trade name of prednisone, 5 mg/tablet, is made by Shandong Xinhua pharmaceutical Co., Ltd, and is the Chinese medicine standard H37020647 which is a positive control group of the experimental western medicine. The prednisone acetate tablets are ground into fine powder and prepared into 1mg/ml suspension by using distilled water for later use.

1.3 Primary reagents and instruments

Complete Freund's adjuvant (Sigma, Lot # SLBZ 9884); incomplete Freund's adjuvant (Sigma, Lot # SLBZ 2765). Guinea pig anti-mouse platelet serum (APS) was prepared by the laboratory with reference to the relevant literature. Mouse TPO, IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma, TGF-beta 1 kits, by Shandong Jiuxiliao Biotechnology Limited. A fully automatic hemocytometer (model number CELL-DYN1700, Abbott, USA); centrifuge (model TGL-168, Shanghai' an pavilion scientific Instrument factory); universal microscope of the DMRBE type (Leica, germany).

2 method

2.1 establishment of ITP mouse model

Reference methods, the GP-APS method was used to establish Immune Thrombocytopenia (ITP) mouse models. The specific method comprises the following steps: taking APS out of a refrigerator at the temperature of-20 ℃, putting the APS in a water bath at the temperature of 56 ℃ for 30min after the APS is completely melted, adsorbing the APS with mouse red blood cells of the same amount of BALB/c for at least 2 times, and adding normal saline according to the ratio of 1: 4-ratio dilution, followed by intraperitoneal injection of 0.1mL diluted APS at 0, 1, 3, 5, 7, 9, 11, 13d, respectively, to maintain a sustained low level of platelets.

2.2 Experimental groups and dosing

50 BALB/C mice were randomly divided into 5 groups, which were a normal control group, a model group, a Qihuang qi-tonifying and blood-engorging high and low dose group, and a prednisone group, 10 mice per group. On day 8 after APS injection, each fraction was administered by intragastric gavage. According to the equivalent dose ratio between human and animal according to the conversion of body surface area, the clinical daily dose of human is 225g crude drug, the equivalent dose of mouse is 9.1 times of human, so the low and high dose groups of Qihuang qi-tonifying and blood-controlling prescription are respectively set to be 30 and 60g/kg, which are respectively 1 and 2 times of the clinical daily dose of human; the daily clinical dosage of prednisone is 1mg/kg, and the administration dosage converted into mice is 9.1mg/kg, and is normalThe group and the model group are administered with the same volume of physiological saline for intragastric administration for 1 time/d and continuous intragastric administration for 14d, and the administration volume is 20 mL/kg ^-1 。

2.3 peripheral blood picture detection

After continuous administration for 14 days, the tail vein of the mouse is bled, diluted in proportion, and a full-automatic blood analyzer is used for detecting the Platelet (PLT) count and the hemoglobin (Hb) value.

2.4Elisa method for detecting serum TPO, IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma and TGF-beta 1 levels

After the administration, the mice are fasted and forbidden to be watered for 8h, the orbital venous plexus is bled, serum is separated, the operation is strictly carried out according to an Elisa kit instruction, and the serum TPO, IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma and TGF-beta 1 levels of each group of mice are detected.

2.5 spleen and thymus indices

Mice were weighed, sacrificed for dissection and isolation of spleen and thymus, dissected free of surrounding adipose tissue, blotted, wet weighed with an electronic balance and recorded, and spleen and thymus indices were calculated: spleen index ═ spleen mass (g)/mouse body weight (g) × 100; thymus index is thymus mass (g)/mouse body weight (g) × 100.

2.6 bone marrow megakaryocyte Classification

Taking a mouse sternum to make a bone marrow smear, dyeing in Reye, counting 100 megakaryocytes under a microscope (1000 x), and classifying the megakaryocytes (including primitive + juvenile megakaryocytes, granular megakaryocytes, platemaking megakaryocytes and naked-nucleus megakaryocytes).

2.7 statistical analysis

Data were processed using SPSS 20.0 statistical software, with data averaged + -SD

Showing that the comparison between groups is performed by analysis of variance, P<A difference of 0.05 is statistically significant.

2 results

2.1 general case observations

The normal group of mice has flexible response, bright hair color and no bleeding point; after the model group mice are injected with APS for 48 hours in the abdominal cavity, the reaction is slow, the hair color is dry, the activity is obviously weakened, and the symptoms of food consumption reduction, skin bleeding and the like can be seen on the 8 th day of model building; after the medicine is taken, the reaction and the activity of the model mouse are obviously increased, the hair color is recovered to be bright, and the bleeding point is obviously reduced.

2.2 Effect of Qi-tonifying and blood-controlling formula of Astragalus membranaceus on PLT count, Hb and TPO level of peripheral blood of mice in ITP model

Compared with the normal group, the peripheral blood PLT count, Hb and TPO level of the mice in the model group are remarkably reduced (P <0.05 or P < 0.01); compared with the model group, the PLT counts and TPO levels of the Qihuang qi-tonifying blood-taking prescription high and low dose groups and the prednisone group are remarkably increased (P is less than 0.01), and the Hb level of the high dose group is remarkably increased (P is less than 0.05); compared with the low dose group, the PLT counts of the high dose group and the prednisone group are obviously increased (P is less than 0.05), and other indexes have no statistical difference (P is more than 0.05). See table 1.

TABLE 1 influence of qi-tonifying and blood-controlling formula of Astragalus membranaceus on PLT count, Hb and TPO levels in ITP model mice: (

n＝10)

Note: in comparison with the normal group, ^* P<0.05， ^** P<0.01; in comparison with the set of models, ^# P<0.05， ^## P<0.01; compared with the low-dose group, the treatment was, ^& P<0.05 (same below).

2.4 influence of Qi-tonifying and blood-controlling formula of Astragalus membranaceus and Astragalus membranaceus on spleen and thymus index of ITP model mouse

Compared with the normal group, the spleen and thymus indexes of the model group are obviously increased (P is less than 0.01); compared with the model group, spleen indexes and thymus indexes of qi-tonifying and blood-controlling formulas of astragalus mongholicus and astragalus membranaceus in the low-dose group and the prednisone group are remarkably reduced (P <0.05 or P < 0.01). See table 2.

TABLE 2 influence of qi-tonifying and blood-controlling formula of Astragalus membranaceus and Astragalus membranaceus on spleen and thymus index in ITP model mice: (

n＝10)

2.3 influence of Qi-tonifying and blood-controlling formula of Qihuang on levels of proinflammatory factors IL-2, IL-6, TNF-alpha and IFN-gamma in serum of ITP model mice

Compared with a normal group, the serum IL-2, IL-6, TNF-alpha and IFN-gamma of the model group mice are remarkably increased (P <0.05 or P < 0.01); compared with the model group, the levels of IL-2, IL-6, TNF-alpha and IFN-gamma of the Qihuang blood-tonifying prescription in the high and low dose groups and the prednisone group are remarkably reduced (P is less than 0.05 or P is less than 0.01); compared with the low dose group, the IL-6 level of only the prednisone group is significantly reduced (P <0.05), the TNF-alpha level of the high dose group and the prednisone group is significantly reduced (P <0.05 or P <0.01), and the rest indexes are not statistically different (P > 0.05). See table 3.

TABLE 3 influence of qi-tonifying and blood-controlling formula on the levels of proinflammatory factors IL-2, IL-6, TNF-alpha and IFN-gamma in serum of ITP model mice ((

n＝10)

2.4 influence of qi-tonifying and blood-controlling formula of Qihuang on serum anti-inflammatory factors IL-4, IL-10 and TGF-beta 1 levels of ITP model mice

Compared with a normal group, the serum IL-4, IL-10 and TGF-beta 1 levels of the mice in the model group are all obviously reduced (P is all less than 0.01); compared with the model group, the levels of IL-4, IL-10 and TGF-beta 1 of the Qihuang qi-tonifying blood-controlling prescription high and low dose groups and the prednisone group are obviously increased (P is less than 0.05 or P is less than 0.01); compared with the low dose group, the IL-4 and TGF-beta 1 levels of the high dose group and the prednisone group are obviously increased (P is less than 0.01), and the IL-10 level of the high dose group is obviously increased (P is less than 0.05). See table 4.

TABLE 4 influence of qi-tonifying and blood-controlling formula of Astragalus membranaceus Hemsl on the levels of IL-4, IL-10 and TGF-beta 1 in serum of ITP model mice ((

n＝10)

2.5 influence of Qi-tonifying and blood-controlling prescription of Astragalus membranaceus on bone marrow megakaryocyte classification of ITP model mouse

Compared with the normal group, the number of the bone marrow plate-producing megakaryocytes of the mice in the model group is obviously reduced (P < 0.01); compared with the model group, the number of bone marrow megakaryocytes of the Qihuang qi-tonifying blood-controlling prescription in the high and low dose groups and the prednisone group is obviously increased (P is less than 0.01). See table 5.

TABLE 5 influence of qi-tonifying and blood-controlling formula of Astragalus membranaceus and Astragalus membranaceus on classification of bone marrow megakaryocytes in ITP model mice: (

n＝10)

Discussion of 3

Immune Thrombocytopenia (ITP) is currently recognized as an autoimmune disorder, the clinical manifestations of which are mainly immune-mediated increased platelet destruction and decreased production. The pathogenesis of ITP is complex and is not clear up to now, and previous researches suggest that the ITP can be caused by humoral immunity and cellular immunity abnormality.

The main pathological change in ITP is peripheral thrombocytopenia, and the injection of exogenous GP-APS can cause platelet destruction. In the research, an ITP mouse model is established by adopting an intraperitoneal injection GP-APS method, and compared with a normal group, the results of the ITP mouse model are compared with those of a model group, the model group mice can show the expressions of food intake reduction, skin bleeding and the like, the PLT count of peripheral blood, TPO and Hb are obviously reduced (P <0.05 or P <0.01), the success of model making of the mouse ITP model is prompted, the spleen and thymus index of the model mice is also obviously increased (P <0.01), and the number of bone marrow plate-producing megakaryocytes is obviously reduced (P < 0.01). After 14 days of continuous treatment by the qi and blood tonifying formula, compared with the model group, the PLT counts and TPO levels of the qi and blood tonifying formula of astragalus mongholicus and the low dose group and the prednisone group are obviously increased (P is less than 0.01), the Hb level of the high dose group is obviously increased (P is less than 0.05), the spleen and thymus indexes are obviously reduced (P is less than 0.05 or P is less than 0.01), and the number of megakaryocytes of the bone marrow producing plates is obviously increased (P is less than 0.01); compared with the low-dose group, the PLT counts of the high-dose group and the prednisone group are obviously increased (P is less than 0.05), and the results indicate that the qi-tonifying and blood-controlling formula of astragalus mongholicus has a good treatment effect on the ITP model mice, and the curative effect of the high-dose group is better than that of the low-dose group.

Research shows that inflammatory cytokines have important significance in maintaining in-vivo immune balance, so the research discusses the pathogenesis of ITP and the action mechanism of the qi-tonifying and blood-taking formula of astragalus mongholicus for treating ITP from the perspective of proinflammatory/inflammation-inhibiting cytokine imbalance. The level of cytokines IL-2, IL-6, TNF-a and IFN-gamma is increased, so that the inflammatory reaction can be aggravated, and the cytokines IL-2, IL-6, TNF-a and IFN-gamma belong to proinflammatory factors; and the increase of IL-4, IL-10 and TGF-beta 1 level can reduce inflammatory reaction, belonging to the inflammation inhibiting factor. IL-2 is an important substance for immune regulation, is mainly secreted by Th1 cells and can enhance the killing activity of T cells, and the research suggests that IL-2 can cause the increase of platelet destruction by promoting the direct cytotoxic effect of the T cells. IL-6 is secreted mainly by Th2 cells, and can promote B cell proliferation, differentiation and antibody secretion, so that the antibody can be combined with the platelet itself, and the platelet is largely destroyed and reduced by the monocyte phagocytosis system, thereby causing ITP. TNF-alpha is a promoter and amplification factor for inflammatory reactions, and can be produced by Th cells, NK cells and B cells. The generation and release of a large amount of TNF-alpha can destroy the immune balance of the organism, and cause the damage of the organism. IFN-gamma is a proinflammatory factor with strong effect, is mainly secreted by Th1 cells, and has inhibitory effect on inflammation-inhibiting factors such as IL-4, TGF-beta and the like. IL-4 is a Th2 type cytokine, has inhibitory action on proinflammatory factors such as IFN-gamma and the like, and has important significance on maintaining the immune balance of organisms. IL-10 exerts immunosuppressive effects mainly through antigen-presenting cells, and particularly has significant inhibitory effects on macrophages and dendritic cells. The research finds that IL-10 can inhibit mononuclear macrophages from releasing immune mediators and reduce the release of inflammatory mediators such as proinflammatory factors TNF-alpha, IL-8 and the like, IL-10 can also inhibit IFN-gamma and TNF-alpha generated by Th1 cells and IL-4 and IL-5 generated by Th2 cells, and IL-10 can inhibit the proliferation of CD4+ T cells and cytokine synthesis to different degrees and inhibit the proliferation of lymphocytes. TGF-beta 1 is a cytokine that regulates inflammatory responses in the body, and regulates the proliferation, differentiation, and activation of immune-related cells such as lymphocytes and macrophages in an autocrine and paracrine manner. TGF-beta 1 can induce Treg cell secretion, and Treg cells play an immunosuppressive role through secreting TGF-beta and IL-10. TGF-beta 1 has negative regulation effect on megakaryocyte development and differentiation process in bone marrow, and can be combined with TGF-beta 1 III type receptor on the surface of megakaryocyte to inhibit mitosis in megakaryocyte nucleus and formation of megakaryocyte colony (CFU-MK), thereby participating in regulation of thrombopoiesis. The research result shows that compared with the normal group, the serum IL-2, IL-6, TNF-alpha and IFN-gamma levels of the model group mice are obviously increased (P is less than 0.05 or P is less than 0.01), and the IL-4, IL-10 and TGF-beta 1 levels are all obviously reduced (P is less than 0.01); after the qi-tonifying and blood-controlling prescription of astragalus mongholicus is given for continuous treatment for 14 days, compared with a model group, the levels of IL-2, IL-6, TNF-alpha and IFN-gamma in serum of a high-dose group, a low-dose group and a prednisone group of the qi-tonifying and blood-controlling prescription of astragalus mongholicus are obviously reduced (P is less than 0.05 or P is less than 0.01), and the levels of IL-4, IL-10 and TGF-beta 1 are obviously increased (P is less than 0.05 or P is less than 0.01); compared with the low dose group, the TNF-alpha level of the high dose group and the prednisone group is remarkably reduced (P <0.05 or P <0.01), the IL-4 and TGF-beta 1 level is remarkably increased (both P <0.01), the IL-6 level of only the prednisone group is remarkably reduced (P <0.05), the IL-10 level of only the high dose group is remarkably increased (P <0.05), and the rest indexes have no statistical difference (P > 0.05). The result indicates that the qi-tonifying and blood-controlling formula of astragalus mongholicus has the effects of reducing proinflammatory factors and improving the level of the inflammation-inhibiting factors on an ITP model mouse, so that the anti-inflammatory effect of the composition is exerted.

During the research, the following are also found: the Chinese medicinal preparation is added with the medicaments for clearing heat and removing toxicity, and activating blood circulation to dissipate blood stasis, which play a key role in adjusting proinflammatory factors and inhibiting secretion imbalance of the inflammatory factors, so that the qi-tonifying and blood-nourishing formula containing astragalus and astragalus has a better clinical treatment effect. In conclusion, the qi-tonifying and blood-taking formula with astragalus mongholicus can increase the levels of bone marrow megakaryocytes and TPO (thermoplastic polyolefin) by regulating the secretion imbalance of proinflammatory factors and inflammation-inhibiting factors of an ITP model mouse, so that the effect of treating ITP is exerted, and a certain dose dependence relationship exists between high and low doses.

It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited thereto, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents can be made in the technical solutions described in the foregoing embodiments, or equivalents thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention. Although the present invention has been described with reference to the specific embodiments, it should be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.

Claims

1. An application of a Chinese medicinal composition in preparing medicine for treating immune thrombocytopenia;

the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 25-35 parts of astragalus membranaceus, 20-30 parts of radix rehmanniae recen, 10-20 parts of poria cocos, 10-20 parts of bighead atractylodes rhizome, 25-35 parts of buffalo horn, 10-15 parts of red peony root, 6-12 parts of cortex moutan, 25-35 parts of hairyvein agrimonia herb and bud, 10-20 parts of madder, 10-20 parts of eclipta, 10-15 parts of selaginella tamariscina, 1-5 parts of pseudo-ginseng powder, 6-12 parts of glabrous sarcandra herb and 3-9 parts of liquorice;

the preparation method of the medicine comprises the following steps:

mixing the above Chinese medicinal compositions, extracting with water, collecting filtrate, concentrating, and drying;

wherein the water extraction times are 2-4 times, the water addition amount is 8-12 times of the total weight of the raw materials, and the water extraction time is 1-2 hours.

2. The use of claim 1, wherein the Chinese medicinal composition is prepared from the following raw materials in parts by weight: 25-30 parts of astragalus membranaceus, 20-25 parts of radix rehmanniae recen, 10-15 parts of poria cocos, 10-15 parts of bighead atractylodes rhizome, 25-30 parts of buffalo horn, 10-12 parts of red peony root, 6-9 parts of cortex moutan, 25-30 parts of hairyvein agrimony, 10-15 parts of madder, 10-15 parts of eclipta, 10-12 parts of selaginella tamariscina, 1-3 parts of pseudo-ginseng powder, 6-9 parts of glabrous sarcandra herb and 3-6 parts of liquorice.

3. The use of claim 1, wherein the Chinese medicinal composition is prepared from the following raw materials in parts by weight: 30-35 parts of astragalus membranaceus, 25-30 parts of radix rehmanniae recen, 15-20 parts of poria cocos, 15-20 parts of bighead atractylodes rhizome, 30-35 parts of buffalo horn, 12-15 parts of red peony root, 9-12 parts of cortex moutan, 30-35 parts of hairyvein agrimony, 15-20 parts of madder, 15-20 parts of eclipta, 12-15 parts of selaginella tamariscina, 3-5 parts of pseudo-ginseng powder, 9-12 parts of glabrous sarcandra herb and 6-9 parts of liquorice.

4. The use of claim 1, wherein the number of aqueous extractions is 2.

5. The use according to claim 1, wherein the amount of water added is 10 times the total weight of the starting materials.

6. The use as claimed in claim 1, wherein the water extraction time is 1 h.

7. Use according to claim 1, wherein the drying is vacuum drying.

8. The use of claim 1, wherein the method further comprises: pulverizing the dried raw materials.