Traditional Chinese medicine composition for treating recurrent abortion and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine compositions, and provides a traditional Chinese medicine composition for treating recurrent abortion and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 2-18 parts of ginseng, 3-30 parts of astragalus membranaceus, 3-30 parts of prepared rehmannia root, 3-24 parts of fructus lycii, 3-24 parts of angelica sinensis, 2-10 parts of radix paeoniae alba, 3-24 parts of schisandra chinensis, 5-40 parts of semen cuscutae, 2-18 parts of ligusticum wallichii, 2-16 parts of rhizoma cyperi, 5-40 parts of raspberry, 3-24 parts of eclipta, 2-20 parts of scutellaria baicalensis, 2-18 parts of polygala tenuifolia and 1-10 parts of liquorice. The traditional Chinese medicine composition is reasonable in compatibility and synergistic in effect, has the effects of reinforcing qi and nourishing blood, tonifying kidney and replenishing essence, and clearing heat and preventing miscarriage, and has an obvious treatment effect on recurrent abortion.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine compositions, in particular to a traditional Chinese medicine composition for treating recurrent spontaneous abortion and application thereof. BACKGROUND

[0002] Recurrent spontaneous abortion (RSA) refers to the continuous occurrence of ≥2 fetal loss (body weight <500g) before 28 weeks of gestation with the same sexual partner. The incidence of RSA in pregnant women is 1%-5%, and the frequency of spontaneous abortion is increasing, and the risk of RSA is also increasing. Traditional Chinese medicine has been recorded in classical works such as Zhuzhibenghoulun and Jinkuiaolue for treating RSA, and traditional Chinese medicine has achieved remarkable effects in treating RSA.

[0003] Recurrent spontaneous abortion is also known as "sliding abortion" or "multiple fetal loss". The premise of pregnancy in women is that the kidney qi is sufficient, the heavenly essence is mature, the qi and blood of the chong and ren channels are harmonious, and the function of the uterus, vessels and collaterals is normal. Any abnormality in the above aspects can lead to sliding abortion. The book Zhuzhibenghoulun·Yunghuizhaofaichou of Chao Yuanfang pointed out: "If the blood and qi are deficient, the uterus is suffering from wind and cold, the blood and qi are insufficient, so it cannot nourish the fetus, so it is multiple fetal loss." The book also puts forward the view that "pregnant women with waist pain like to have abortions." The main pathogenesis of recurrent spontaneous abortion is mainly: (1) deficiency of kidney qi, and the fetus loses the system: traditional Chinese medicine believes that the kidney stores essence and is the root of life, and the kidney is the main system of the fetus. If the parents are not enough in the preconception, the organs are weak, although the two essences are combined, but the fetus cannot be real, it is easy to cause repeated pregnancy and repeated abortion. Or after pregnancy, improper sexual behavior, deficiency of kidney qi, instability of chong and ren, and inability to system the fetus, the fetus loses the nourishment, and thus the fetus is lost; (2) deficiency of spleen and stomach, deficiency of qi and blood: spleen and stomach are the root of life, and the source of qi and blood. If the spleen and stomach are insufficient, the blood is deficient and the qi is weak, the chong and ren blood pool is empty, or the spleen is injured by diet and fatigue, the qi and blood are deficient, or the disease is long and the qi and blood are consumed, the qi and blood are deficient, the chong and ren are not stable, and repeated pregnancy and repeated abortion lead to sliding abortion; (3) blood stasis, chong and ren are not smooth: if the mother has a tumor, the blood stasis in the vessels and organs, the chong and ren are damaged, the blood is not smooth, and the kidney is damaged by long-term blood stasis, the fetus is not nourished, and the fetus is lost; (4) liver depression and qi stagnation, blood heat and other factors can lead to recurrent spontaneous abortion.

[0004] At present, the main treatment of recurrent spontaneous abortion in western medicine is cell therapy, immunosuppressants, progesterone, anticoagulants and other therapies. Traditional Chinese medicine treats recurrent spontaneous abortion with the principles of tonifying the kidney and invigorating the spleen, tonifying qi and blood, regulating the chong and ren, and preventing and treating the damage, and pays attention to "prevention first, combination of prevention and treatment". Traditional Chinese medicine can prevent and treat recurrent spontaneous abortion by regulating oxidative stress, promoting decidualization, regulating immune function, promoting embryo invasion, improving endometrial receptivity, promoting angiogenesis, and affecting autophagy, and has broad development prospects in clinical application. SUMMARY

[0005] Therefore, the present application aims to provide a traditional Chinese medicine composition for treating recurrent miscarriage and application thereof to solve the problems mentioned in the background.

[0006] To achieve the above-mentioned purposes, the technical scheme of the present application is as follows:

[0007] A traditional Chinese medicine composition for treating recurrent miscarriage is prepared from the following raw medicinal materials by weight:

[0008] Radix ginseng 2-18 parts, Radix astragali 3-30 parts, Radix rehmanniae preparata 3-30 parts, Fructus lycii 3-24 parts, Radix angelicae sinensis 3-24 parts, Radix paeoniae alba 2-10 parts, Schisandra chinensis 3-24 parts, Semen cuscutae 5-40 parts, Rhizoma chuanxiong 2-18 parts, Rhizoma cyperi 2-16 parts, Rubus chingii 5-40 parts, Herba ecliptae 3-24 parts, Radix scrophulariae 2-20 parts, Polygala 2-18 parts, and Radix glycyrrhizae 1-10 parts.

[0009] Further, the traditional Chinese medicine composition is prepared from the following raw medicinal materials by weight:

[0010] Radix ginseng 9 parts, Radix astragali 15 parts, Radix rehmanniae preparata 15 parts, Fructus lycii 12 parts, Radix angelicae sinensis 12 parts, Radix paeoniae alba 10 parts, Schisandra chinensis 12 parts, Semen cuscutae 20 parts, Rhizoma chuanxiong 9 parts, Rhizoma cyperi 8 parts, Rubus chingii 20 parts, Herba ecliptae 12 parts, Radix scrophulariae 10 parts, Polygala 9 parts, and Radix glycyrrhizae 5 parts.

[0011] Further, the preparation method of the traditional Chinese medicine composition comprises the following steps:

[0012] a. taking Radix ginseng, Semen cuscutae, and Rubus chingii, crushing them into fine powder, sieving, and reserving;

[0013] b. taking Radix scrophulariae, Radix angelicae sinensis, Rhizoma chuanxiong, Schisandra chinensis, and Rhizoma cyperi, refluxing extraction with 6-12 times of 60%-90% ethanol for 1-3 times, each time for 1-3 hours, filtering, combining the filtrates, recovering ethanol, and concentrating into thick paste with relative density of 1.30-1.35 measured at 60℃, reserving;

[0014] c. taking the rest of Radix astragali, Radix rehmanniae preparata, Fructus lycii, Radix paeoniae alba, Herba ecliptae, Polygala, and Radix glycyrrhizae, decocting with 6-12 times of water for 1-3 times, each time for 1-3 hours, filtering, combining the filtrates, standing, taking supernatant, and concentrating into thick paste with relative density of 1.30-1.35 measured at 60℃, reserving;

[0015] d. combining a, b, and c, mixing, drying, and crushing into fine powder, to obtain the traditional Chinese medicine composition.

[0016] Further, a, taking Radix ginseng, Semen cuscutae, and Rubus chingii, crushing them into fine powder, sieving, and reserving;

[0017] b. Scutellaria, Angelica, Chuanxiong, Schisandra, Cyperus, 8 times the amount of 70% ethanol reflux extraction 2 times, 2 hours each time, filter, combined filtrate, recovery of ethanol, concentrated into 60 ℃ measured relative density of 1.30~1.35 thick paste, ready for use;

[0018] c. The rest of the Astragalus, Rehmannia, Medlar, white peony root, Herba Phryta, Polygala, licorice seven, plus 8 times the amount of water decoction 2 times, 2 hours each time, filter, combined filtrate, stand, take the supernatant, concentrated into 60 ℃ measured relative density of 1.30~1.35 thick paste, ready for use;

[0019] d. The above a, b, c, combined, mixed, dried, pulverized into fine powder, the traditional Chinese medicine composition is obtained.

[0020] Further, according to the conventional method is made into pharmaceutically acceptable traditional Chinese medicine preparation.

[0021] Further, the pharmaceutical dosage form of the traditional Chinese medicine preparation is tablet, capsule, granule or pill.

[0022] Another object of the present application is to provide a kind of traditional Chinese medicine composition for treating recurrent abortion in the preparation of drug for treating recurrent abortion.

[0023] Compared with the prior art, the present application has the following advantages:

[0024] The traditional Chinese medicine composition for treating recurrent abortion and its application of the present application, by reasonable compatibility of each component, mutual synergistic effect, has the effect of benefiting qi and nourishing blood, tonifying kidney and filling essence, clearing heat and securing fetus, the traditional Chinese medicine composition of the present application can significantly reduce the whole blood viscosity and plasma viscosity of blood stasis syndrome model rats; increase the red blood cell deformation ability, inhibit the aggregation of red blood cells; improve the blood circulation of the body, increase the blood supply and oxygen supply of tissues. It has obvious improvement effect on the hemorheological characteristics of blood stasis syndrome "thick, sticky, coagulation, aggregation"; the number of embryos absorbed by pregnant rats of the traditional Chinese medicine composition of the present application is significantly reduced, the number of surviving embryos is significantly increased, and the embryo absorption rate of pregnant rats is significantly reduced, which has obvious therapeutic effect on recurrent abortion. BRIEF DESCRIPTION OF DRAWINGS

[0025] The drawings constituting a part of the present application are used to provide further understanding of the present application, the schematic embodiments of the present application and their descriptions are used to explain the present application, and do not constitute improper limitation on the present application. In the drawings:

[0026] Figure 1 The effect of the traditional Chinese medicine composition for treating recurrent abortion described in the embodiments of the present application on the embryo development of abortion model pregnant rats and the structure of maternal-fetal interface;

[0027] Figure 2The effect of the traditional Chinese medicine composition for treating recurrent miscarriage described in this embodiment of the invention on serum inflammatory factors in a miscarriage model of pregnant mice. Detailed Implementation

[0028] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0029] In the description of this invention, it should be noted that the terms "upper," "lower," "inner," and "back," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0030] The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0031] Example 1

[0032] This embodiment relates to a traditional Chinese medicine composition for treating recurrent miscarriage and its application. The traditional Chinese medicine composition is made from the following raw materials in parts by weight:

[0033] Ginseng 2-18 parts, Astragalus 3-30 parts, Rehmannia glutinosa 3-30 parts, Lycium barbarum 3-24 parts, Angelica sinensis 3-24 parts, Paeonia lactiflora 2-10 parts, Schisandra chinensis 3-24 parts, Cuscuta chinensis 5-40 parts, Ligusticum chuanxiong 2-18 parts, Cyperus rotundus 2-16 parts, Rubus idaeus 5-40 parts, Eclipta prostrata 3-24 parts, Scutellaria baicalensis 2-20 parts, Polygala tenuifolia 2-18 parts, Glycyrrhiza uralensis 1-10 parts.

[0034] Example 2

[0035] The preparation method of the traditional Chinese medicine composition in Example 1 includes the following steps:

[0036] a. Take ginseng, dodder seed, and raspberry, grind them into a fine powder, sift them, and set aside;

[0037] b. Take Scutellaria baicalensis, Angelica sinensis, Ligusticum chuanxiong, Schisandra chinensis, and Cyperus rotundus, and extract them by reflux with 6 to 12 times the amount of 60 to 90% ethanol 1 to 3 times, each time for 1 to 3 hours. Filter, combine the filtrates, recover the ethanol, and concentrate into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use.

[0038] c. Add 6 to 12 times the amount of water to the remaining seven herbs: Astragalus membranaceus, Rehmannia glutinosa, Lycium barbarum, Paeonia lactiflora, Eclipta prostrata, Polygala tenuifolia, and Glycyrrhiza uralensis, decoct 1 to 3 times, 1 to 3 hours each time, filter, combine the filtrates, let stand, take the supernatant, and concentrate it into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use.

[0039] d, combine a, b, and c above, mix well, dry, and pulverize into fine powder to obtain the traditional Chinese medicine composition.

[0040] The traditional Chinese medicine composition is prepared into a pharmaceutically acceptable traditional Chinese medicine preparation according to a conventional method. Specifically, the medicinal dosage form of the traditional Chinese medicine preparation is a tablet, a capsule, a granule, or a pill.

[0041] Example 3

[0042] The traditional Chinese medicine composition is prepared from the following raw medicinal materials by weight:

[0043] Radix ginseng 9 parts, Radix astragali 15 parts, Radix rehmanniae preparata 15 parts, Fructus lycii 12 parts, Radix angelicae sinensis 12 parts, Radix paeoniae alba 10 parts, Schisandra chinensis 12 parts, Semen cuscutae 20 parts, Rhizoma chuanxiong 9 parts, Rhizoma cyperi 8 parts, Rubus chingii 20 parts, Herba ecliptae 12 parts, Radix scrophulariae 10 parts, Polygala 9 parts, and Radix glycyrrhizae 5 parts.

[0044] Example 4

[0045] The preparation method of the traditional Chinese medicine composition includes the following steps:

[0046] a, pulverize Radix ginseng, Semen cuscutae, and Rubus chingii into fine powder, sieve, and reserve;

[0047] b, extract Radix scrophulariae, Radix paeoniae alba, Fructus lycii, Radix angelicae sinensis, and Rhizoma chuanxiong with 8 times the amount of 70% ethanol by reflux extraction for 2 times, 2 hours each time, filter, combine the filtrates, recover ethanol, and concentrate into a thick paste with a relative density of 1.30-1.35 measured at 60°C, and reserve;

[0048] c, decoct the remaining Radix astragali, Radix rehmanniae preparata, Fructus lycii, Radix paeoniae alba, Herba ecliptae, Radix scrophulariae, and Radix glycyrrhizae with 8 times the amount of water for 2 times, 2 hours each time, filter, combine the filtrates, stand, take the supernatant, and concentrate into a thick paste with a relative density of 1.30-1.35 measured at 60°C, and reserve;

[0049] d, combine a, b, and c above, mix well, dry, and pulverize into fine powder to obtain the traditional Chinese medicine composition.

[0050] Example 5

[0051] The traditional Chinese medicine composition is prepared from the following raw medicinal materials by weight:

[0052] Radix ginseng 90g, Radix astragali 150g, Radix rehmanniae preparata 150g, Fructus lycii 120g, Radix angelicae sinensis 120g, Radix paeoniae alba 100g, Schisandra chinensis 120g, Semen cuscutae 200g, Rhizoma chuanxiong 90g, Rhizoma cyperi 80g, Rubus chingii 200g, Herba ecliptae 120g, Radix scrophulariae 100g, Polygala 90g, and Radix glycyrrhizae 50g.

[0053] Prepare according to the following steps:

[0054] a. Take three kinds of ginseng, dodder, raspberry, etc. Grind into fine powder, sieve, and reserve;

[0055] b. Take five kinds of Huangqin, Danggui, Chuanqiong, Wuweizi, Xiangfu, etc. Use 8 times the amount of 70% ethanol to reflux extract 2 times, 2 hours each time, filter, combine the filtrate, recover the ethanol, and concentrate into thick paste with relative density of 1.30-1.35 (60°C measurement), and reserve;

[0056] c. The remaining seven kinds of Huangqi, Shudihuang, Gouqi, Baishao, Muhuanglian, Yuanzhi, and Gancao are added with 8 times the amount of water to decoct 2 times, 2 hours each time, filter, combine the filtrate, stand, take the supernatant, and concentrate into thick paste with relative density of 1.30-1.35 (60°C measurement), and reserve;

[0057] d. Combine the above a, b, and c, mix well, dry, grind into fine powder (100 mesh), and obtain the composition.

[0058] Example 6

[0059] Clinical test of the traditional Chinese medicine composition of the present application for treating recurrent miscarriage

[0060] I. Acute toxicity test of the traditional Chinese medicine composition on rats

[0061] SPF level rats were used, half male and half female, weighing 198.4±11.4g, animal license number: SCXK-(Jing)2021-0011, purchased from Beijing Vito Lihua Experimental Animal Technology Co., Ltd.

[0062] Test method

[0063] 1. Quarantine and domestication of animals After purchasing the animals, first pre-cage them, set a temporary pre-cage number, and measure the weight on the purchase day. Take the purchase day as the first day, and the quarantine and domestication period is 2 days. During this period, animals with abnormalities in appearance, general state, and possible impact on the test are excluded during the test.

[0064] 2. Preparation of test sample Before the test, the test sample is weighed with a ten-thousandth balance, and the maximum concentration is prepared with distilled water.

[0065] 3. Dose, method, frequency, and duration of administration of test sample and control sample

[0066] 3.1 Route and method of administration: oral administration.

[0067] 3.2 Dose design reason: According to the results of the pre-test, the maximum dose volume of 20mL / kg was used for gavage in rats, and 4 animals were given gavage 3 times a day. No death occurred. Therefore, the maximum dose method was used in this test.

[0068] Based on the preliminary test results, with a maximum oral dose volume of 20 ml / kg and a maximum oral concentration of 65.3 g / 100 ml for rats, administered three times a day, no deaths occurred in any of the four animals. Therefore, this experiment adopted the maximum dose method.

[0069] 3.3 Dosage frequency and timing: Administer 3 times a day, with an interval of 5 hours.

[0070] 4. Detection methods and frequencies for various indicators

[0071] 4.1 Observation of general symptoms

[0072] 4.1.1 Observation frequency: Observe continuously for 4 hours after administration, and then observe and record once in the morning and once in the afternoon every day for 14 consecutive days.

[0073] 4.1.2 Observation of all experimental animals after drug administration.

[0074] 4.1.3 Observation content: Observe the animal's coat color, mental state, spontaneous activities, respiration, oral and nasal secretions, diet, urine and feces, as well as the time of death and recovery.

[0075] 4.2 Weight Measurement

[0076] 4.2.1 The determination method was carried out using a "KF-H2 electronic balance".

[0077] 4.2.2 All animals were purchased before administration and all test animals were tested after administration.

[0078] 4.2.3 Number of measurements: Measured once on the day of purchase, once on the day of animal grouping (day 1 of drug administration), once every day after drug administration, and once on the day of dissection.

[0079] 4.3 Anatomy

[0080] 4.3.1 The dissection was conducted the day after the experiment ended.

[0081] 4.3.2 Number of dissections: All experimental animals after drug administration.

[0082] 4.3.3 Dissection Method: After euthanizing the animal by cervical dislocation, a dissection is performed. During the dissection, the animal's external appearance and all organs are visually observed. If any abnormalities are found, a histopathological examination is conducted.

[0083] 5. Statistical methods used in data analysis: Statistical analysis was performed using SPSS statistical software. Measurement results are expressed as mean ± standard deviation. ± S) represents the mean; analysis of variance was used to compare quantitative data between groups.

[0084] Test results

[0085] No abnormal reaction was observed in the 20 rats within 14 days after administration, and no animal died. The animals were sacrificed after 14 days, and no abnormality was observed in the dissection of organs. Body weight change: During the observation period, the body weight of the surviving animals gradually increased, as shown in Tables 1 and 2.

[0086] Table 1. Effect of the Chinese medicine composition on the body weight of rats (♂) ± S)

[0087]

[0088] Note: The number in the brackets is the number of surviving animals P>0.05

[0089] Table 2. Effect of the Chinese medicine composition on the body weight of rats (♀) ± S)

[0090]

[0091] Note: The number in the brackets is the number of surviving animals P>0.05

[0092] The experimental results showed that no obvious acute toxicity reaction was observed when the maximum gavage dose of 39.18 / kg was administered 3 times a day.

[0093] Second, the effect of the Chinese medicine composition on improving the hemorheology of blood stasis model rats

[0094] 1. Experimental materials

[0095] 1.1 Experimental animals

[0096] Strain: Wistar rats, grade: SPF level, number: 40, gender: 20 males and 20 females, body weight: 250-270g. License number: SCXK-(Jing)2021-0011, purchased from Beijing Vantianlihua Experimental Animal Technology Co., Ltd.

[0097] 1.2 Drugs and reagents

[0098] Adrenaline hydrochloride injection, product batch number: 2007201. Suicheng Pharmaceutical Co., Ltd. Heparin sodium, batch number: 20241202, Shanghai Huishi Biochemical Reagent Co., Ltd. Isoflurane, batch number: 22102301, Ruivode Life Science Co., Ltd.

[0099] 1.3 Experimental instruments

[0100] MR animal anesthesia machine, model: Matrx VIP3000 series isoflurane (ether) anesthesia volatile tank (Isoflurane Vaporizer), product serial number: V1547492, MIDMARK company of the United States. Blood rheology tester, model: BT-300, Beijing Bolai Medical Technology Co., Ltd. Table type high speed low temperature centrifuge, model: Biofuge Stratos, Heraeus company of Germany. Electronic balance: specifications: PRACTUM313-1CN, serial number: 0028892121, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0101] 2 Experimental method

[0102] 2.1 Experimental grouping

[0103] After the experimental animals were purchased, they were first stabilized and adapted to the environment for 1 week. Then they were randomly divided into: normal control group; blood stasis model group; this traditional Chinese medicine composition is divided into high dose, medium dose and low dose of 3 different dosing groups, a total of 5 groups, 8 animals in each group, 4 males and 4 females.

[0104] 2.2 Administration method

[0105] The traditional Chinese medicine composition was prepared into 53.6%, 26.8% and 13.4% three different concentrations of suspension liquid with the daily drinking water of rats; the administration dose of the traditional Chinese medicine composition was high dose: 5.36g / kg / d, medium dose: 2.68g / kg / d, low dose: 1.34g / kg / d. Different dosing groups of animals used the same gavage volume method to give different drug concentrations, and the gavage volume was 10ml / kg / d. The normal control group and the model control group of rats were given the same volume (10ml / kg / d) of daily drinking water of rats. The body weight of rats was measured once every 7 days, and the administration dose was adjusted according to the body weight change. The activity, food intake and water intake of rats after administration were observed daily.

[0106] 2.3 Establishment of blood stasis model

[0107] The compound stasis factors of subcutaneous injection of adrenaline plus ice water immersion stimulation were used to establish the blood stasis model of rats. 1 hour after the last administration, except for the normal control group, the rest of the animals were subcutaneously injected (sc) with adrenaline hydrochloride injection 1mg / kg body weight. After 2 hours, the animals were immersed in 4℃ ice water for 5 minutes and taken out. After 2 hours, the same dose (1mg / kg body weight) of adrenaline hydrochloride injection was subcutaneously injected again to establish the blood stasis model of rats.

[0108] 2.4 Blood sample processing

[0109] After the blood stasis model of rats was established, the rats were fasted for 16 hours without water. The rats were placed in the closed induction box of the VMR animal anesthetizing machine, and the rats were anesthetized by adjusting the volatile release flow of anesthetic isoflurane through the control knob. 4 ml of blood was taken from the abdominal aorta, and 0.1% heparin sodium was used for anticoagulation. 1 ml of whole blood was taken for different shear rate whole blood viscosity detection; the rest of the anticoagulated blood was centrifuged at 3000 rpm for 15 min, and 1 ml of plasma was separated for plasma viscosity detection.

[0110] 2.5 Detection index

[0111] 2.5.1 Whole blood viscosity

[0112] 1 ml of anticoagulated whole blood was taken to detect high shear rate (100 / S), medium shear rate (30 / S) and low shear rate (3 / S) whole blood viscosity with a blood rheometer.

[0113] 2.5.2 Plasma viscosity

[0114] 1 ml of plasma was taken to detect plasma viscosity with a blood rheometer.

[0115] 2.6 Statistical analysis method

[0116] SPSS 20.0 statistical software was used for data analysis, and the experimental results were expressed as mean ± standard deviation (S). One-way ANOVA test was used for comparison between different groups, and LSD test was selected when the variance was equal, and Dunnett T3 test was selected when the variance was not equal. When P <0.05, it indicated that there was significant difference.

[0117] 3 Experimental results

[0118] The experimental results showed that the traditional Chinese medicine composition could significantly reduce the whole blood viscosity and plasma viscosity of blood stasis model rats; increase the deformation ability of red blood cells, inhibit the aggregation of red blood cells; improve the blood circulation of the machine, and increase the blood and oxygen supply. It has obvious improvement effect on the hemorheology characteristics of blood stasis syndrome such as "thick, sticky, coagulation and aggregation". The detailed results are shown in Table 3.

[0119] Table 3. Effect of the traditional Chinese medicine composition on whole blood viscosity, plasma viscosity and hematocrit of acute blood stasis model rats ±s, n=8

[0120]

[0121] Note: Compared with the model control group: ** P<0.01; * P<0.05.

[0122] Three, the therapeutic effect of the traditional Chinese medicine composition on abortion model mice ​

[0123] 1. Experimental materials

[0124] 1.1 Experimental animals

[0125] Strain: ICR mice, grade: SPF level, gender: female, age: 8 weeks, number: 48. License number: SCXK-(Jing)2021-0011, purchased from Beijing Vantoll Life Science and Technology Co., Ltd.

[0126] 1.2 Drugs and reagents

[0127] Lipopolysaccharide (LPS), S11060, MLOT: KS373982, BR, derived from Escherichia coli, Shanghai Yuanye Bio-technology Co., Ltd. Hydroxyprogesterone caproate tablets (Oxytocin), batch number: T001-250201B, Yangzhou Auctate Pharmaceutical Co., Ltd. Adrenaline hydrochloride injection, batch number: 2411251, Tianjin He Ping Pharmaceutical Co., Ltd. Hydroxyurea tablets, batch number: 5A0023DG0, drug standard number H37021289, Qilu Pharmaceutical Co., Ltd. Isoflurane, batch number: G45880, product of Ruivode Life Science Co., Ltd. Isoflurane, product batch number: 24081502, product of Ruivode Life Science Co., Ltd. TNF-α (product number: JL10484), IL-10 (product number: JL20242), IL-1β (product number: JL18442) were purchased from Shanghai Jianglai Biological Technology Co., Ltd.

[0128] 2. Experimental methods

[0129] 2.1 LPS-induced mouse abortion model

[0130] The experiment was performed on the 10th day of pregnancy by injecting LPS 0.2 mL (0.1 μg) per mouse via the tail vein to establish a mouse abortion model, and the samples were taken on the 14th day of pregnancy.

[0131] 2.2 Experimental grouping

[0132] 48 ICR normal pregnant mice were selected and randomly divided into normal pregnancy group, abortion model group, dydrogesterone group, and traditional Chinese medicine composition high, medium and low dose groups, 8 mice in each group, a total of 6 groups.

[0133] 2.3 Administration method

[0134] Preventive administration from the 0th day of pregnancy, gavage administration, once a day in the morning, administration for 2 weeks. The traditional Chinese medicine composition is prepared into 39.0%, 19.5% and 9.8% three different concentrations of suspensions with the daily drinking water of mice respectively; the positive control drug medroxyprogesterone tablets (Oxin Tong) is prepared into a suspension of 52 mg / 100 ml with the daily drinking water of mice. The administration dose of the traditional Chinese medicine composition is high dose: 7.8 g / kg / d, medium dose: 3.9 g / kg / d, low dose: 2.0 g / kg / d; the administration dose of the positive control drug medroxyprogesterone tablets is 10.4 mg / kg / d. According to the surface area equivalent dose conversion coefficient between different animals, the animals in different administration dose groups are administered with different drug concentrations with the same gavage volume, and the gavage volume is 20 ml / kg / d. The normal pregnancy group and the abortion model control group of mice are given the same volume (20 ml / kg / d) of the daily drinking water of mice. The body weight of the mice is weighed once every 3 days, and the administration dose is adjusted according to the body weight change. The activity, food intake and water drinking of the mice after administration are observed daily.

[0135] 2.4 Detection index

[0136] 2.4.1 HE staining of the maternal-fetal interface

[0137] The morphology of the maternal-fetal interface of the pregnancy abortion model mice is observed by HE staining method. The uterus of the pregnant mice is collected and fixed in 4% paraformaldehyde for 12-16 h. After dehydration of the tissue sample, paraffin embedding is performed, and continuous sectioning is performed with a section thickness of 4 μm. The cross section of the maternal-fetal interface tissue is cut. Histological examination and analysis are performed under an optical microscope.

[0138] 2.4.2 ELISA detection

[0139] The content of IL-10, TNF-α and IL-1β in the serum of the mice is detected by enzyme-linked immunosorbent assay (ELISA). The setting, sample addition, plate sealing, incubation, plate washing, incubation, reaction termination and color development are performed according to the instructions of the detection kit. The optical density (OD value) of each well is measured by an enzyme label instrument at a wavelength of 450 nm. Then, the regression equation of the standard curve is calculated according to the concentration of the standard sample and the measured OD value. Finally, the OD value of the sample is substituted into the equation to calculate the concentration of the measured sample.

[0140] 2.4.3 Observation of the uterine cavity of pregnant mice, embryo state and calculation of embryo resorption rate

[0141] After the pregnant mice are sacrificed by carbon dioxide euthanasia at 14 d of pregnancy, the abdominal cavity is opened, and the uterus is fully exposed. The growth of the uterus is observed, and the number of embryo implantation and the number of embryo resorption are recorded. The embryo resorption rate is calculated as follows: embryo resorption rate (%) = resorbed embryo number / (resorbed embryo number + survived embryo number) x 100%

[0142] 2.5 Statistical processing

[0143] SPSS 20.0 statistical software was used for data analysis, and the experimental results were expressed as mean ± standard deviation (±s). One-way ANOVA test was used for comparison between different groups, LSD test was selected when the variance was equal, and Dunnett T3 test was selected when the variance was not equal. When P<0.05, it indicates significant difference.

[0144] 3 Experimental results

[0145] 3.1 Effects of the Chinese medicine composition on the embryonic development of the abortion model pregnant mice and the structure of the maternal-fetal interface

[0146] The HE staining results show that the Control group: the placental tissue structure is complete, the decidua tissue and the trophoblast cell distribution are orderly; the interface area is clear, the interface boundary is clear; the trophoblast structure is clear, the blood sinus structure is complete and open, and no obvious inflammatory infiltration and tissue necrosis is observed. The Model group: the decidua area structure is obviously destroyed, the cell components are disordered; the interface area is fuzzy, the interface boundary disappears; the trophoblast structure is disordered, part of the blood sinus is narrow, and local cellulose-like deposition and reticular necrosis can be observed, accompanied by cellulose-like exudation and neutrophil infiltration. The Dydrogesterone group: the placental decidua tissue is orderly distributed, and tissue disintegration can be observed at the placental base; the interface area is clear, the interface boundary is clear; the trophoblast structure is slightly disordered, the blood sinus structure is complete and open, and no obvious cellulose-like deposition and reticular necrosis is observed locally. The high-dose group (YXG-H group) of the present application: the placental decidua tissue is slightly disordered; the interface area is clear, the interface boundary is clear; the trophoblast structure is clear, the blood sinus structure is complete and open, and obvious cellulose-like deposition and reticular necrosis can be observed locally. The medium-dose group (YXG-M group) of the present application: the placental decidua tissue is orderly distributed; the interface area is slightly fuzzy, the interface boundary disappears; the trophoblast structure is slightly disordered, the blood sinus structure is complete and open, and no obvious cellulose-like deposition and reticular necrosis is observed locally. The low-dose group (YXG-L group) of the present application: the placental decidua tissue structure is slightly disordered; the interface area is clear, the interface boundary is clear; the trophoblast structure is slightly disordered, part of the blood sinus is narrow, and a small amount of cellulose-like deposition and reticular necrosis can be observed locally. The results are shown in Figure 1 .

[0147] 3.2 Effects of the Chinese medicine composition on the serum inflammatory factors of the abortion model pregnant mice

[0148] ​Compared with the Control group, the serum IL-1β, TNF-α levels and TNF-α / IL-10 ratio in the Model group were significantly increased, and the IL-10 level was decreased (P<0.01 or P<0.05, respectively). Compared with the Model group, the serum IL-1β, TNF-α levels and TNF-α / IL-10 ratio in the dydrogesterone, the high and medium dose groups of the Chinese medicine composition (YXG-H, YXG-M groups) were significantly decreased, and the IL-10 level was significantly increased (P<0.01 or P<0.05, respectively); the TNF-α / IL-10 ratio in the low dose group of the Chinese medicine composition (YXG-L group) was significantly decreased, and the IL-10 level was significantly increased (P<0.05, respectively). The results showed that the Chinese medicine composition might inhibit the serum pro-inflammatory factors IL-1β, TNF-α levels, and regulate the anti-inflammatory factor IL-10 level to the normal range. It should be noted that compared with the model group, *P<0.05 (significant difference); **P<0.01 (very significant difference); ns. P>0.05 (no significant difference). The results are shown in Figure 2 (Wherein, ±s, n=8).

[0149] 3.3 Effect of the Chinese medicine composition on the embryo resorption rate of the abortion model pregnant rats

[0150] 1) Observation results: In the normal pregnancy control group, the embryos were similar in size, shaped like a string of beads, evenly distributed, bright red in color, with clear blood vessels, no congestion or bleeding, and visible red cord-like placenta. In the abortion model control group, the absorbed or dead embryo rats were observed, and there were obvious old congestion or blood clot embryos in the uterus, with dark red color, even brown or black purple, significantly reduced embryo volume, and atrophic and absorbed blastocyst, and some embryos disappeared.

[0151] 2) Embryo resorption rate: Compared with the normal pregnancy control group, the number of absorbed embryos in the abortion model control group was significantly increased, the number of surviving embryos was significantly reduced, and the embryo resorption rate was significantly increased, with statistically significant difference (P<0.01); compared with the abortion model group, the number of absorbed embryos in the pregnant rats of the Chinese medicine composition at 7.8 g / kg, 3.9 g / kg and 2.0 g / kg dose groups (YXG-H, YXG-M groups) was significantly reduced, the number of surviving embryos was significantly increased, and the embryo resorption rate of the pregnant rats was significantly reduced, with statistically significant difference (P<0.01 or P<0.05, respectively). The experimental results are shown in Table 4.

[0152] Table 4. Effect of the Chinese medicine composition on the embryo resorption rate of the abortion model pregnant rats ±s, n=8)

[0153]

[0154] Note: Compared with the model group,* P<0.05; ** P<0.01.

[0155] The above experimental results show that the traditional Chinese medicine composition can significantly reduce the whole blood viscosity and plasma viscosity of blood stasis syndrome model rats; increase the red blood cell deformation ability, inhibit the red blood cell aggregation; improve the blood circulation, increase the tissue blood supply and oxygen supply. It has obvious improvement effect on the hemorheology characteristics of blood stasis syndrome "thick, sticky, coagulation and aggregation"; the number of embryos absorbed by the pregnant rats in the three dose groups of the traditional Chinese medicine composition is significantly reduced, the number of surviving embryos is significantly increased, and the embryo absorption rate of the pregnant rats is significantly reduced, so the traditional Chinese medicine composition has obvious therapeutic effect on recurrent miscarriage.

[0156] The traditional Chinese medicine composition for treating recurrent miscarriage and its application adopting the above implementation solution is guided by the theory of traditional Chinese medicine, and the effects of multiple drugs are matched, the monarch, minister, assistant and messenger play their respective roles, the qi is supplemented and the blood is nourished, the kidney is tonified and the essence is filled, the heat is cleared and the fetus is protected, the symptoms and root causes are treated, the root cause is treated as the main treatment, and the characteristics of multiple targets and multiple mechanisms are jointly played. Animal experiments show that the drug has definite curative effect, small side effect and high cure rate. The drug better meets the medication needs of patients. Compared with similar technologies, it has obvious technical progress.

[0157] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A traditional Chinese medicine composition for treating recurrent spontaneous abortion, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ginseng 2-18 parts, Astragalus 3-30 parts, Rehmannia glutinosa 3-30 parts, Lycium barbarum 3-24 parts, Angelica sinensis 3-24 parts, Paeonia lactiflora 2-10 parts, Schisandra chinensis 3-24 parts, Cuscuta chinensis 5-40 parts, Ligusticum chuanxiong 2-18 parts, Cyperus rotundus 2-16 parts, Rubus idaeus 5-40 parts, Eclipta prostrata 3-24 parts, Scutellaria baicalensis 2-20 parts, Polygala tenuifolia 2-18 parts, Glycyrrhiza uralensis 1-10 parts.

2. The traditional Chinese medicine composition for treating recurrent spontaneous abortion according to claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: Ginseng 9 parts, Astragalus 15 parts, Rehmannia glutinosa 15 parts, Lycium barbarum 12 parts, Angelica sinensis 12 parts, Paeonia lactiflora 10 parts, Schisandra chinensis 12 parts, Cuscuta chinensis 20 parts, Ligusticum chuanxiong 9 parts, Cyperus rotundus 8 parts, Rubus idaeus 20 parts, Eclipta prostrata 12 parts, Scutellaria baicalensis 10 parts, Polygala tenuifolia 9 parts, Glycyrrhiza uralensis 5 parts.

3. The traditional Chinese medicine composition for treating recurrent miscarriage according to any one of claims 1, characterized in that: The preparation method of the traditional Chinese medicine composition includes the following steps: a. Take ginseng, dodder seed, and raspberry, grind them into a fine powder, sift them, and set aside; b. Take Scutellaria baicalensis, Angelica sinensis, Ligusticum chuanxiong, Schisandra chinensis, and Cyperus rotundus, and extract them by reflux with 6 to 12 times the amount of 60% to 90% ethanol 1 to 3 times, each time for 1 to 3 hours. Filter, combine the filtrates, recover the ethanol, and concentrate into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use. c. Add 6 to 12 times the amount of water to the remaining seven herbs: Astragalus membranaceus, Rehmannia glutinosa, Lycium barbarum, Paeonia lactiflora, Eclipta prostrata, Polygala tenuifolia, and Glycyrrhiza uralensis, decoct 1 to 3 times, 1 to 3 hours each time, filter, combine the filtrates, let stand, take the supernatant, and concentrate it into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use. d. Combine items a, b, and c above, mix well, dry, and pulverize into a fine powder to obtain the Chinese herbal composition.

4. The traditional Chinese medicine composition for treating recurrent miscarriage according to claim 3, characterized in that: a. Take ginseng, dodder seed, and raspberry, grind them into a fine powder, sift them, and set aside; b. Take Scutellaria baicalensis, Angelica sinensis, Ligusticum chuanxiong, Schisandra chinensis and Cyperus rotundus, and extract them twice with 8 times the amount of 70% ethanol, each time for 2 hours. Filter, combine the filtrates, recover the ethanol, and concentrate them into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use. c. Add 8 times the amount of water to the remaining seven herbs: Astragalus membranaceus, Rehmannia glutinosa, Lycium barbarum, Paeonia lactiflora, Eclipta prostrata, Polygala tenuifolia, and Glycyrrhiza uralensis, decoct twice for 2 hours each time, filter, combine the filtrates, let stand, take the supernatant, and concentrate it into a thick paste with a relative density of 1.30 to 1.35 measured at 60℃, for later use. d. Combine items a, b, and c above, mix well, dry, and pulverize into a fine powder to obtain the Chinese herbal composition.

5. The traditional Chinese medicine composition for treating recurrent miscarriage according to claim 1, characterized in that: Traditional Chinese medicine preparations are manufactured using conventional methods to achieve pharmaceutically acceptable results.

6. The traditional Chinese medicine composition for treating recurrent miscarriage according to claim 1, characterized in that: The pharmaceutical dosage forms of the traditional Chinese medicine preparations are tablets, capsules, granules, or pills.

7. The traditional Chinese medicine composition for treating recurrent miscarriage according to any one of claims 1-6, and its use in the preparation of a drug for treating recurrent miscarriage.

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