CN114668800A

CN114668800A - Traditional Chinese medicine composition for treating female hyperplasia of mammary glands and preparation method and application thereof

Info

Publication number: CN114668800A
Application number: CN202210545779.XA
Authority: CN
Inventors: 杨建华; 胡君萍; 居博伟
Original assignee: First Affiliated Hospital of Xinjiang Medical University
Current assignee: First Affiliated Hospital of Xinjiang Medical University
Priority date: 2022-05-19
Filing date: 2022-05-19
Publication date: 2022-06-28

Abstract

The invention discloses a traditional Chinese medicine composition for treating hyperplasia of mammary glands, which is prepared from the following raw material medicines in parts by weight: 6-10 parts of clematis root, 6-18 parts of papermulberry fruit and 6-12 parts of tangerine seed. The main medicine of the invention cuts the pathogenesis of mammary nodules in the composing prescription, the three medicines and the pathogenesis threads are buckled to mutually supplement each other to play the role, and the effect on treating the hyperplasia of mammary glands is obvious.

Description

Traditional Chinese medicine composition for treating female hyperplasia of mammary glands and preparation method and application thereof

Technical Field

The invention particularly relates to a traditional Chinese medicine composition for treating female hyperplasia of mammary glands and a preparation method and application thereof.

Background

Mammary Gland Hyperplasia (MGH) is a pathological state of abnormal Hyperplasia of Mammary tissue, usually Hyperplasia of Mammary epithelium and fibrous tissue, and abnormal quantity and morphology of Mammary structures caused by Hyperplasia and involution insufficiency of different degrees, not tumors, but inflammations; the main pathological manifestations of the disease are typical pregnancy-like changes of mammary gland principal and stroma and tumorous cystic hyperplasia, the clinical manifestations are breast mass, unilateral or bilateral breast distending pain or tenderness, accompanied by menstrual disorder, emotional changes, part of patients merging nipple discharge and the like, and the disease is one of the mammary gland diseases with the highest incidence.

According to the clinical symptoms of hyperplasia of mammary glands in the traditional Chinese medicine, the hyperplasia of mammary glands is classified into the category of nodules of breast, which is seen in the middle Tibetan Jing at the earliest time, and doctors in different generations consider that hyperplasia of mammary glands is closely related to disharmony between Chong and ren vessels and stagnation of liver qi caused by deficiency of kidney qi. The book Sheng Ji Zong Lu states that qi is obstructed and stagnated, or the wind is pathogenic, the qi is hard and swollen, and the pain is nucleated. The "breast nodules" in the "orthodox department of surgery" indicates that the breast nodules are nodules of breast milk, like the shape of a pill egg, or weigh down and cause pain, or do not cause pain, the skin color is unchanged, and the nodules are relieved and grown with joy. In addition, doctors think that women are "littley and mostly unsmooth" and "partial littley negative", and "love and jealousy worry" in the aspect of sexual conditions, so "often complain about" women are prone to unsmooth depression of the liver due to emotional depression, and qi stagnation and blood stasis appear in women. Therefore, the traditional medicine considers that the mammary gland hyperplasia is influenced by a coordination mechanism of kidney-Chong and ren-uterus-breast, which is consistent with the discovery of the modern medicine that the growth, development and secretion function of the mammary gland are regulated by a hypothalamus-pituitary-ovary axis, so that the mammary gland hyperplasia disease is found in liver and kidney, stagnation of liver qi, obstruction of qi activity, deficiency of liver and kidney and disharmony of Chong and ren meridians can further cause the generation and development of the mammary gland hyperplasia.

At present, the traditional Chinese medicine treatment for hyperplasia of mammary glands is provided with the prescriptions of bupleurum root liver-soothing powder, modified Xiaoyao powder, nodules of breast, cassia twig and poria pill, modified prescription, modified bupleurum root liver-soothing powder and the like. The western medicine treatment of hyperplasia of mammary glands mainly comprises hormone and surgical treatment, and hormone inhibitors such as aromatase inhibitor (tamoxifen) are mainly adopted clinically; for patients with unobvious medical conservative treatment effect, surgical resection treatment is mostly adopted clinically. However, western medicine treatment usually mainly takes hormone drugs, has many adverse reactions, is easy to cause nausea, vomiting and amenorrhea and increases the risk of endometrial cancer, and is easy to cause major body trauma in operation, and mammary gland hyperplasia is diffuse and easy to recur, and the operation treatment also brings high economic burden to patients; the traditional Chinese medicine is usually made of compound recipe, but the traditional Chinese medicine has the common characteristics of both the classical ancient recipe and the compound recipe for treating the hyperplasia of mammary glands in the prior art: the traditional Chinese medicine composition has a large number of medicinal materials, for example, the bupleurum liver-soothing powder has 7 medicines, the modified Xiaoyao powder has 10 medicines, the nodules of breast is more than 10 medicines, part of the medicinal materials are rare medicinal materials, the cost is high, the research mechanism is complex and cannot be clarified, and the difficulty is increased for later-period achievement transformation.

Disclosure of Invention

In order to solve the problems, the invention provides a traditional Chinese medicine composition for treating hyperplasia of mammary glands, which is prepared from the following raw material medicines in parts by weight:

6-10 parts of clematis root, 6-20 parts of papermulberry fruit and 6-12 parts of tangerine seed.

Further, the feed additive is prepared from the following raw materials in parts by weight:

6 parts of clematis root, 6-18 parts of papermulberry fruit and 6-12 parts of tangerine seed.

6 parts of clematis root, 12 parts of papermulberry fruit and 12 parts of tangerine seed.

Further, the tangerine seeds are salt tangerine seeds.

Furthermore, the traditional Chinese medicine composition is a preparation prepared by taking medicinal powder of raw material medicines, or water or organic solvent extracts of the raw material medicines as active ingredients and adding pharmaceutically acceptable auxiliary materials.

Further, the preparation is an oral preparation, and the oral preparation is granules, paste, powder, pills or solution.

The invention also provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps:

(1) weighing the raw materials according to the proportion;

(2) grinding the raw materials into powder, or extracting with water or organic solvent, and adding common adjuvants or auxiliary components.

The invention finally provides application of the traditional Chinese medicine composition in preparing a medicine for treating hyperplasia of mammary glands.

The traditional Chinese medicine composition for treating hyperplasia of mammary glands consists of Clematis root, fructus broussonetiae and salt tangerine seed, wherein the Clematis root belongs to dried roots and rhizomes of Clematis chinensis Osbeck of Ranunculaceae, and has the effects of dispelling wind-damp, dredging channels and collaterals and the like. The Li Shizhen specifically describes the efficacy of the clematis root: 'Xuan Tong five internal organs, removing cold stagnation in abdomen, heart and diaphragm phlegm, long-term accumulation in the abdomen, and mass of the liver-wind and spleen-qi'. Fructus Broussonetiae is dry mature fruit of Vent, Broussonetia papyrifera (L.) of Moraceae, and has sweet taste and enters liver and kidney meridians. It is recorded in Yi Zong jin Jian that Broussonetia papyrifera (L.) Nees has effects of nourishing yin, tonifying liver and kidney, and invigorating yang qi. The salt tangerine seeds are dry mature seeds of Citrus reticulata Blanco of Rutaceae and cultivated varieties thereof, and are salted by general rules of Chinese pharmacopoeia to obtain the salt tangerine seeds which are bitter and neutral in taste, enter liver and kidney channels, and have the effects of regulating qi, relieving pain, dissipating stagnation, dispelling cold and the like.

According to the traditional Chinese medicine, the deficiency of liver and kidney and the imbalance of Chong and ren channels are the root cause of the hyperplasia of mammary glands, the stagnation of qi due to depression of the liver and the stagnation of qi and the blood stasis due to phlegm coagulation are the targets of the hyperplasia of mammary glands, phlegm and fluid stay in the breast due to the deficiency of the liver and the kidney, and finally, the lump is formed, accumulated and obstructed, and the pain is caused if the lump is obstructed. In the composition, clematis root has the effects of dredging channels and collaterals, softening and resolving hard mass and dispelling phlegm, fructus broussonetiae has the effects of tonifying liver and kidney, regulating and tonifying Chong and ren meridians, softening and resolving hard mass, is a monarch drug together, and salt tangerine seed has the effects of regulating qi to alleviate pain, dispelling cold and resolving hard mass and enhancing the effect of treating hyperplasia of mammary glands.

Animal tests and clinical tests prove that the invention changes syndrome differentiation into disease differentiation aiming at the characteristics of breast pain and mass of a patient which are eliminated and grown along with emotional changes; aiming at the nodules of breast formed by disharmony of Chong and conception vessels, the treatment follows the principle of reaching the depressed people and consolidating and dispersing, and the compound obtained by compatibility has obvious effect on treating the female breast hyperplasia caused by liver and kidney deficiency and disharmony of Chong and conception vessels, has no toxic or side effect, high safety and low cost, and has practical popularization and application values.

Compared with the traditional Chinese medicine prescription for treating hyperplasia of mammary glands in the prior art, the invention has the following remarkable progress:

1. the invention finds out the basic pathogenesis of the hyperplasia of mammary glands, namely deficiency of liver and kidney and disharmony of thoroughfare and conception vessels, treats the basic pathogenesis and hits the target point, and really realizes that the diseases are treated by drugs and the drugs are combined, thereby having quick effect. Breast pain symptoms were reduced within three doses and disappeared within 1 week; with the disappearance of pain, the lump in the breast disappeared about 4 weeks. For consolidation of curative effect, it is usually used for 8 weeks. Syndrome differentiation is carried out during disease differentiation, and the interference of concurrent symptoms on syndrome types is avoided aiming at the basic pathogenesis of the hyperplasia of mammary glands.

2. In the formula, the clematis root has the effects of relaxing the channels and collaterals, dispelling phlegm and fluid, breaking hardness and eliminating mass, dispersing stagnant qi, softening hardness and dissolving stasis, softening and eliminating breast lumps and also having the functions of relieving pain and resisting inflammation, the paper mulberry fruit has the effects of tonifying liver and kidney, benefiting essence and blood, dissolving nodules, dredging and reducing blood, regulating Chong and ren meridians, the salt tangerine seeds have the effects of soothing the liver and regulating qi, relieving pain and dispelling cold, so the clematis root, the paper mulberry fruit and the salt tangerine seeds can be used together to achieve the curative effect of treating both symptoms and root causes, and the three medicines are mutually reinforced and play the role of achieving stasis and dispersing stasis. The main medicine of the invention is used for cutting the pathogenesis of mammary nodules in the composing prescription, and the three medicines are screwed with the pathogenesis threads to supplement each other to play the role. The invention has precise and definite formula principle and obvious treatment effect.

3. The three medicinal materials in the invention have wide sources, are easy to obtain, have lower cost and have sufficient substance guarantee.

Based on the traditional Chinese medicine theory, the 'syndrome differentiation and treatment' and the 'holistic concept' are important theoretical bases and effective material sources for research and development of the traditional Chinese medicine compound preparation, and are unique advantages of the traditional Chinese medicine compound preparation.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1 is a B-ultrasonic diagram of mammary gland of rats in each group

FIG. 2 is the staining pattern of the mammary gland HE of each group of rats

FIG. 3 is a Masson staining chart of mammary gland of each group of rats

FIG. 4 shows the pre-treatment hormone levels in a patient

FIG. 5 shows the hormone levels after treatment of a patient

FIG. 6 is a B-ultrasonic breast view of patient 1 before treatment

FIG. 7 is a B-ultrasonic photograph of the mammary gland of patient 1 after treatment

FIG. 8 is a B-ultrasonic view of the mammary gland before treatment of patient 2

FIG. 9 is a B-ultrasonic breast view of patient 2 after treatment

FIG. 10 is a B-ultrasonic photograph of the mammary gland before treatment of patient 3

FIG. 11 patient 3 post-treatment mammary B-ultrasonography

FIG. 12 is the index chart of liver and kidney function before treatment

FIG. 13 is the hepatorenal function index chart of the patients after treatment

FIG. 14 is the electrocardiogram index of the patient

FIG. 15 is a chart of the patient's blood routine before treatment

FIG. 16 is a graph of the patient's 3 blood routine index using the optimization method of the present invention

FIG. 17 is a urine routine index of a patient

Detailed Description

EXAMPLE 1 the drug of the invention

The formula is as follows: clematis root 6g and papermulberry fruit 20 g.

The preparation method comprises the following steps:

soaking the two raw materials in water for 1 hour, controlling the water to submerge the medicine by 2 cm when the medicine is decocted, timing when the water is boiled, decocting for 30 minutes, taking out 150ml of decoction after the decoction is finished, adding water into the decocting pot again, controlling the water to submerge the medicine by 1 cm, carrying out secondary decoction, timing when the water is boiled, decocting for 20 minutes again, taking out 150ml of decoction, combining the decoctions, and mixing uniformly. The medicine is taken twice in the morning and at night, and the interval between the two times of the medicine taking is 8 hours.

The using method comprises the following steps: the medicine is taken 1 dose a day in the morning and at night, and the two doses are separated by 8 hours.

Example 2 the drug of the invention

The formula is as follows: 6g of clematis root, 6g of fructus broussonetiae and 6g of salt tangerine seed.

The preparation method comprises the following steps:

mixing the three raw materials in proportion, soaking the mixture in water for 1 hour, controlling the water to submerge the medicine for 2 cm when the medicine is decocted, timing when the water is boiled, decocting for 30 minutes, taking out 150ml of decoction after the decoction is finished, adding water into the decocting pot again, controlling the water to submerge the medicine for 1 cm, carrying out secondary decoction, timing when the water is boiled, decocting for 20 minutes again, taking out 150ml of decoction, combining the decoctions, and uniformly mixing. The medicine is taken twice in the morning and at night, and the interval between the two times of the medicine taking is 8 hours.

Example 3 the drug of the invention

Clematis root 6g, papermulberry fruit 12g and salt tangerine seed 6 g.

The preparation method comprises the following steps:

soaking the three raw materials in water for 1 hour, controlling the water to submerge the medicine by 2 cm when the medicine is decocted, timing when the water is boiled, decocting for 30 minutes, taking out 150ml of decoction after the decoction is finished, adding water into the decocting pot again, controlling the water to submerge the medicine by 1 cm, carrying out secondary decoction, timing when the water is boiled, decocting for 20 minutes again, taking out 150ml of decoction, combining the decoctions, and uniformly mixing. The medicine is taken twice in the morning and at night, and the interval between the two times of the medicine taking is 8 hours.

Example 4 the invention

The formula is as follows: clematis root 6g, papermulberry fruit 12g and salt tangerine seed 12 g.

The preparation method comprises the following steps:

soaking the three raw materials in water for 1 hour, controlling the water to submerge the medicines by 2 cm when the medicines are decocted, timing when the water is boiled, decocting for 30 minutes, taking out 150ml of decoction after the decoction is finished, then adding water into the decocting pot again, controlling the water to submerge the medicines by 1 cm, carrying out secondary decoction, timing when the water is boiled, decocting for 20 minutes again, taking out 150ml of decoction, and combining and uniformly mixing the two decoctions. .

EXAMPLE 5 the inventive Agents

The formula is as follows: clematis root 6g, papermulberry fruit 18g and salt tangerine seed 6 g.

The preparation method comprises the following steps:

soaking the three raw materials in water for 1 hour, controlling the water to submerge the medicine by 2 cm when the medicine is decocted, timing when the water is boiled, decocting for 30 minutes, taking out 150ml of decoction after the decoction is finished, adding water into the decocting pot again, controlling the water to submerge the medicine by 1 cm, carrying out secondary decoction, timing when the water is boiled, decocting for 20 minutes again, taking out 150ml of decoction, combining the decoctions, and uniformly mixing.

The using method comprises the following steps: the medicine is taken 1 dose a day in the morning and at night, and the interval between the two doses is 8 hours.

The advantageous effects of the present invention are further illustrated by the following test examples

Test example 1 animal test study

The invention aims to screen out a compound with obvious effect of treating the hyperplasia of mammary glands by means of biochemical indexes, pathological tissue observation, imaging detection and other technologies and by carrying out pharmacodynamic investigation on different proportions of the clematis root, the paper mulberry fruit and the salt tangerine seed through an animal model.

1. Animal model establishment and drug intervention

1.1 medicaments

Clematis root (Hunan Qing pure science and technology Co., Ltd., lot No. 2012001C), paper mulberry fruit (Hunan Qing pure science and technology Co., Ltd., lot No. 1908001S), salt orange seed (Hunan Qing pure science and technology Co., Ltd., lot No. 2012001H). Estradiol benzoate for veterinary use (Shanghai Quanyu science and technology animal pharmaceuticals Co., Ltd., batch No. 210301); progesterone injection (Zhejiang Xianju pharmaceutical Co., Ltd., lot number: 33020828); tamoxifen (Yangzhou pharmaceutical industry group Co., Ltd., batch No.: 20072191).

1.2 animals

SPF-grade female SD non-pregnant rats aged 9 weeks and 180-; the animals are all raised in an animal experiment center of Xinjiang medical university, and in the experiment period, the animals can freely drink and eat water, the room temperature is 20-25 ℃, the humidity is 40-70%, the illumination period is light and dark, each time is 12 hours, and the ventilation is good.

1.3 methods

Referring to the draft prepared by the Chinese medicine society of medicine "preparation of mammary gland hyperplasia animal models", rats are injected with estradiol benzoate intramuscularly at 0.5mg/kg/d for 1 time (qd) for 25 days continuously, and then progesterone is injected intramuscularly at 4mg/kg/d for 1 time (qd) for 5 days continuously. The structure and the form of the mammary tissue of the animal are observed under a light mirror after the model is built, and the mammary acinar hyperplasia is selected as the main judgment basis for the success of the model building according to the pathological diagnosis standard in the preparation standard of the mammary hyperplasia animal model.

After successful modeling, 80 rats were randomly divided into 8 groups by body mass, namely a normal group, a model group, a positive control group, a basic formula (prepared according to example 1) intervention group, a preferred formula 1 (prepared according to example 2) intervention group, a preferred formula 2 (prepared according to example 3) intervention group, a preferred formula 3 (prepared according to example 4) intervention group and a preferred formula 4 (prepared according to example 5) intervention group. The preferred dose of the prescription and the basic prescription rat should be 2.34g/kg/d (the dose is calculated according to the conversion factor of human and rat being 6.3 and the clinical crude drug amount of the basic prescription is 26g/70 kg/d; the calculation process is that the clinical human dose is 26g/70kg/d which is 371.4mg/kg/d, 6.3 x 371.4mg/kg/d which is 2340mg/kg/d which is 2.34g/kg/d), the positive control group is administrated with tamoxifen, the dose is 5mg/kg/d, the normal group and the model group are perfused with physiological saline for 1 time every day, and the normal group and the model group are perfused with physiological saline for 30 days continuously. After the rat is subjected to the last administration, fasting and water prohibition are carried out for 12 hours, and mammary gland B-ultrasonic imaging examination is carried out; after the end of the imaging examination, rats were anesthetized by intraperitoneal injection of 10% chloral hydrate. Biochemical detection of blood: taking blood from abdominal aorta, 12000r/min, centrifuging for 15min, taking serum, freezing and storing at-80 ℃, using the serum in a subsequent kit for detecting blood biochemistry and ELISA (enzyme-linked immunosorbent assay) to detect hormone level, and inspecting blood biochemistry and hormone level change of each group. Sampling and observing indexes of mammary gland specimen: the 2 nd and 3 rd pairs of bilateral mammary glands of the rats are taken down, the pathological change of the mammary gland tissue of each group of rats is observed under an optical microscope, and the mammary fibrosis condition is observed by Masson staining. Taking down the uterus of the rat, measuring the diameter of the uterus by using a vernier caliper, weighing the uterus by using an electronic balance, and calculating the uterus index; and the rat heart, liver, spleen, lung, kidney were removed and weighed.

2. Index detection

(1) Breast diameter and height

The diameter and height of the udder of each group of rats were precisely measured with a vernier caliper, and the difference between the groups was compared.

(2) Uterine diameter measurement

The diameter of the uterus was measured with a vernier caliper and the diameter of the uterus was compared for each group.

(3) Weighing of organs

The heart, liver, spleen, lung, kidney and uterus were weighed accurately using an analytical balance, and the organ index and uterus index were calculated.

(4) Biochemical index of blood

And (5) detecting by using a biochemical kit.

Reduced Glutathione (GSH), superoxide dismutase (SOD), and Malondialdehyde (MDA).

ELISA measures serum hormone levels.

Estradiol (E)₂) Progesterone (P)_g) Prolactin (PRL), Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH) levels.

(5) Pathological examination of the breast

HE staining: the mammary gland is put into 4% paraformaldehyde solution, and dehydrated by conventional 95% ethanol. Paraffin embedding, slicing, HE staining, and observing the arrangement of ductal epithelium of the mammary gland of each group of rats, the number of acini in ductal cavity, acinus cavity, lobule and lobule after HE staining of the mammary tissue section of the rat by an optical biological microscope.

Masson staining: the mammary gland is put into 4% paraformaldehyde solution, and dehydrated by conventional 95% ethanol. Paraffin embedding, sectioning, staining according to Masson staining kit: dyeing with a prepared Weiert hematoxylin staining solution for 6min, differentiating with an acidic ethanol differentiation solution for 10s, washing with distilled water for 1min, returning blue to a Masson blue staining solution for 5min, washing with distilled water for 1min, dyeing with a ponceau fuchsin staining solution for 6min, washing with 1% glacial acetic acid for 1min, washing with a phosphomolybdic acid solution for 1-2min, adding aniline blue for dyeing for 2min, washing with 1% glacial acetic acid for 1min, storing at room temperature after transparence and sealing, and observing the fat pad, acinus number, catheter arrangement and fibrosis condition of rat mammary gland tissue sections after Masson staining by a biological microscope.

2.3 mammary B-ultrasonic

Adopting a Canon ultrahigh frequency ultrasonic instrument (Aplioi800TUS), using a probe with the model of (i24LX8), preparing skin of a rat in advance, unhairing, fixing the four limbs of the rat, keeping the rat quiet and still, lying down, fully exposing a nipple, uniformly coating an ultrasonic coupling agent on the probe, and displaying the complete shape of the mammary gland of the rat by a display screen of the ultrasonic instrument, thus acquiring an image.

3. Results

3.1 diameter and height of breasts of groups of rats

After the rats are anesthetized, the rats are fixed, the heights and the diameters of the breasts of each group of rats are accurately measured by using a vernier caliper and are subjected to statistical analysis, the heights of the breasts of each group have statistical differences, and compared with a basic formula intervention group, the heights and the diameters of the breasts of the rats after the optimization formula intervention are obviously reduced, wherein the improvement of the optimization formula 3 is more obvious.

(see Table 1).

TABLE 1 Effect of the invention on the Breast height and diameter of various groups of rats

Note:^#p compared with normal control group<0.01，^*Comparison with model group p<0.05，^**Comparison with model group p<0.01，^△Comparison with the basic equation p<0.05。

3.2 visceral and uterine indices

The visceral organs of the rats are slowly stripped and taken down, the weight of each visceral organ is accurately weighed by an analytical balance, the diameter of the uterus is accurately measured by a vernier caliper, compared with a model group, the index of each visceral organ of a drug intervention group has no obvious difference, each group of uterus index has statistical difference, and compared with a basic formula intervention group, the uterus index of the rats after the intervention of an optimization formula is obviously improved. (see tables 2 and 3).

TABLE 2 Effect of the invention on the organ index of rats of each group

Note:^*comparison with model group p<0.05。

TABLE 3 Effect of the invention on uterine index and diameter of various groups of rats

3.3 oxidative stress indicator detection

Compared with the model group, the levels of SOD and GSH in the blood serum of the rat are obviously increased after the drug intervention, the MDA level is obviously reduced, and the statistical difference (P is less than 0.01) is provided, while compared with the basic formula intervention group, the oxidative stress level unbalance of the rat after the optimization formula intervention is obviously improved, wherein the improvement of the optimization formula 3 is the most obvious.

(see Table 4)

TABLE 4 Effect of the invention on serum oxidation factor of rats in various groups

Note: # comparison with Normal control group p<0.01. comparison of p with model group<0.01，^△Comparison with the basic equation p<0.05。

3.4ELISA for serum hormone levels

Compared with the model group, the serum E of the rats in the Chinese medicinal intervention group is given₂PRL and LSH are significantly reduced, P_gAnd LH was significantly elevated, with statistical differences in the hormone levels (P) in the rats of each group<0.01) and the imbalance in hormone levels in rats after the intervention of the optimization formula was significantly improved compared to the basic formula intervention group, with the most significant improvement of optimization formula 3 (see table 5).

TABLE 5 Effect of the invention on the serum hormone levels in various groups of rats

Note:^#p compared with normal control group<0.01，^**Comparison with model group p<0.01，^△Comparison with the basic equation p<0.05。

3.5 mammary gland B Superknot fruit Observation

The rat mammary gland B suprajunction fruit showed that the mammary nodules were significantly reduced in rats given the group of the invention compared to the model group (see figure 1 for details).

3.6 Observation of mammary gland histopathology

Fixing with 4% formaldehyde, selecting fixed specimen, treating with tissue dehydrator, embedding paraffin to obtain paraffin block, cutting into 3mm tissue slices, sealing with neutral gum after HE staining, and observing animal mammary gland tissue structure and shape under light mirror.

The normal group has no hyperplasia of mammary glands, slight fibrosis of mammary gland tissues and normal and orderly and compact arrangement of fat cell forms, the model group has slight to moderate hyperplasia of mammary glands, mild to severe dilatation of acinar cavities and mild to moderate secretion of acinar, and secretion can be seen in a mammary duct, the fibrosis of the mammary gland tissues is severe, and fat cells are ruptured and arranged disorderly; compared with the model group, the hyperplasia of mammary glands of the basic formula group, the optimization formula 1 group, the optimization formula 2 group, the optimization formula 3 group and the optimization formula 4 group are reduced to different degrees, and the hyperplasia of mammary glands is represented by the number of lobules of mammary glands, the diameter and the number of acini, the endocrine of glandular cavities is obviously reduced, the fibrotic tissues are reduced, and the fat cells are restored to be complete and are arranged closely. (see fig. 2 and 3).

4. Conclusion

The traditional Chinese medicine compound can obviously improve the mammary gland pathological changes of a mammary gland hyperplasia model rat by integrating the B-ultrasonic examination and pathological results of mammary glands; good regulation of GSH, SOD, MDA and E in mammary gland serum of rat with mammary gland hyperplasia₂、P_gPRL, LSH, LH levels, thereby reducing proliferative injury of mammary tissue in rats; statistical analysis is carried out on organ coefficients, uterine indexes and uterine diameters, and as a result, the compound of the invention has no significant difference compared with normal groups, which indicates that the traditional Chinese medicine compound is safe and nontoxic.

Animal experiment results also show that the optimization formula added with the salt orange kernels can improve the pathological changes of the mammary glands of model animals, wherein the optimization formula 3 (the ratio is 1:2:2) has obvious advantages in the aspects of improving the pathological changes of the mammary glands of the model animals, regulating and controlling the oxidative stress level, the hormone level and the like, so that the optimization formula 3 (the ratio is 1:2:2) is selected for clinical application practice, and theoretical support is provided for the development of new traditional Chinese medicines.

Test example 2 clinical test

1. Clinical data

30 mammary gland hyperplasia patients who meet the case selection standard in clinic of the fourth subsidiary hospital of Xinjiang medical university from 2021 to 2021 within 12 months are treated, the age of the patients is 24-50 years, the average age is 30.16 years, the course of disease is 3 months-4 years, the average age is 1.9 +/-1.2 years, the age distribution of the patients is reasonable, the range of the course of disease is wide, and the requirements of the research are met. The medicine taking method comprises the following steps: the traditional Chinese medicine compound is administered to 30 patients, and the administration lasts for 4 weeks. Observation indexes are as follows: the change of breast pain and hyperplasia lumps of mammary glands before and after treatment, the total integral of symptom signs, the integral of pain and the integral of accompanying symptoms take 4 weeks as an observation time phase; serum estradiol (E) for pre-and post-menstrual luteal phases₂) Progesterone (P), Prolactin (PRL). Meanwhile, the safety of the medicine is evaluated by detecting liver function, kidney function and heart function.

1.1 diagnostic criteria

Refer to "surgery" for diagnostic criteria related to hyperplasia of mammary glands. (Chenkouchi, Wangjianpin. science [ M ].8 edition. Beijing. people's health Press, 2013.254-255).

1.2 inclusion criteria

(1) Meets the diagnosis standard of hyperplasia of mammary glands in "surgery".

(2) Women, age between 18 and 55 years.

1.3 exclusion criteria

(1) Gestation and lactation. (2) Mastitis, and mammary gland malignant tumor. (3) All hormone drugs are taken. (4) Patients with serious heart, liver and kidney diseases. (5) Patients with mental disease.

1.4 differentiation of the syndromes according to the Chinese medicine

The main symptoms are: local pain in the breast; the lump boundary of the part of the breast is not clear and the texture is tough; the tongue tip has nodule spots, dark red or purple tongue, and even coarse and swollen sublingual collaterals;

the secondary symptoms are as follows: the relationship between local focus or mass of breast and menstrual cycle or mood is not obvious; the pulse is wiry, slippery or astringent. Chest harmony, oppression and irritability, heaviness and lassitude.

1.5 evaluation index and examination of safety

General physical examination: before treatment, at the end of treatment week 4, the examination and recording were carried out once.

Blood and urine routine: before treatment, check once at the end of treatment week 4.

Liver function, kidney function, electrocardiogram: before treatment, check once at the end of treatment week 4.

1.6 efficacy scores

Chinese medicine symptom scoring: before treatment, at the end of treatment week 4, the examination and recording were carried out once.

B ultrasonic examination: before treatment, at the end of treatment week 4, the examination and recording were carried out once.

Hormone level checking: before treatment, at the end of treatment week 4, the examination and recording were carried out once.

2. Method of treatment

On the basis of the animal efficacy experiment results, the composition of the traditional Chinese medicine compound disclosed by the invention, namely the composition of example 4 (clematis root: papermulberry fruit: salt tangerine seed: 1:2:2), is selected by a clinical test treatment group, and is taken warm twice in the morning and at night, so that the curative effect is compared with that before and after taking the medicine.

3. Observation of Chinese medicine syndrome integral

3.1 scoring criteria:

refer to the clinical research guiding principle of new Chinese medicine for treating cystic hyperplasia of breast. Breast pain scoring is performed using a recognized Visual Analogue Scale (VAS) pain scoring standard. 0 is no pain; 1-3 are mild pain, which can be tolerated by patients; 4-6 are classified as painful and affecting sleep, and patients can tolerate it; 7-10 points out that the patient has a gradually intensified pain feeling, the pain of the patient is intolerable, and the appetite and the sleep are influenced. The patient himself/herself circles the pain rating on the corresponding number.

0 minute: none;

and 2, dividing: pain is dull pain or scurrying pain, which stops working when going;

and 4, dividing: pain with tenderness and obvious distending pain;

6 min: pain is either a drop or prick with marked tenderness.

Breast mass size scoring:

0 minute: none;

and 2, dividing: the unilateral or bilateral breast lumps are single small lumps, or are granular and have small range, or are rope objects, are softer and are limited in one quadrant;

and 4, dividing: the mass is large, or is in the shape of a sheet or a disc, which affects the breasts, but the range is in two quadrants;

6 min: large mass, multiple cystic nodules, involvement of double breasts, in more than two quadrants.

Chest and rib distending pain:

0 minute: none;

1 minute: slight chest and costal distending pain, stopping in time;

and 2, dividing: the chest and rib distending pain can be tolerated and persists;

and 3, dividing: the distending pain of the chest and the rib is hard to endure.

Dysphoria and irritability:

0 minute: none;

1 minute: impatience occasionally occurs;

and 2, dividing: irritability and anger;

and 3, dividing: restlessness and irritability make it difficult to control itself.

After 4 weeks of treatment, the Chinese medicine syndrome scores before and after treatment, including breast pain score and breast mass (size, texture) score, were recorded.

4. Determination of therapeutic effect

Refer to the guideline of clinical research of new Chinese medicine. The efficacy index (n) ═ i [ (pre-treatment integral-post-treatment integral) ÷ pre-treatment integral ] × 100%.

(1) The clinical cure is as follows: the breast pain disappears and the breast lumps disappear after the treatment, and the syndrome integral value is 0;

(2) the effect is shown: after treatment, the integral value of the traditional Chinese medicine symptoms is reduced by more than or equal to 70 percent, or the length and the diameter of the tumor are reduced by more than half;

(3) the method has the following advantages: after treatment, the integral value of the traditional Chinese medicine symptoms is reduced by more than or equal to 30 percent and less than 70 percent, or the length and the diameter of the tumor are reduced by more than one third;

(4) and (4) invalidation: those not meeting the above standards.

Total effective rate (clinical recovery + significant effect + effective)/total number of cases x 100%.

5. Results of clinical studies

The effect of this study on the symptoms and hormone levels of patients with mammary gland hyperplasia, the levels of symptoms of patients with mammary gland hyperplasia before and after treatment were statistically different (see table 6), the levels of estrogen, progestin and prolactin were statistically different, and the levels of follitropin and luteinizing hormone were indistinguishable. (see table 7 and figures 4-5), the treatment effect of the hyperplasia of mammary glands is shown in tables 8-9, and B-ultrasonic pictures of mammary glands before and after treatment of part of patients are shown in figures 6-11.

TABLE 6 patient symptom statistics

Note:^*comparison with before treatment P<0.05；^**Comparison with before treatment P<0.01。

TABLE 7 peripheral blood E of patients treated with the present invention₂P, PRL, LSH and LH content effects

Note:^*comparison with before treatment P<0.05。

Table 8 effective statistics of pain and mass in patients with the present invention

TABLE 9 results of treatment of patients with the present invention

From the above results it can be seen that: the compound of the invention obviously improves the symptoms of breast distending pain, mass size, mood fluctuation, chest and hypochondrium swelling and stuffiness, dysphoria, testiness, heavy body, lassitude, dark red tongue and the like of a patient, has obvious significance on the breast distending pain, and has obvious significance on the difference before and after administration through statistical treatment, thereby showing that the compound of the invention has obvious effect on hyperplasia of mammary glands and can effectively improve the life quality of the patient.

After the compound of the invention is applied, the compound of the invention is proved to have obvious improvement effect on B-ultrasonic examination of human, which shows that the compound of the invention can improve the breast lesion of a patient with hyperplasia of mammary glands.

The compound of the invention can improve the hyperplasia degree of mammary gland tissues and can adjust E in human serum₂P, PRL, LSH, LH hormone level expression.

6. Significance and result of safety detection of the invention

6.1 liver function 5 indexes and their selection significance

Alanine Aminotransferase (ALT): the ALT is mainly present in various cells, the content of the liver cells is the most, and a large amount of ALT is released into blood when various viral hepatitis in the acute stage and drug toxic liver cells are necrotic, so the ALT is an important index for diagnosing viral hepatitis and toxic hepatitis; is a sensitive marker of acute hepatocyte damage.

Aspartate Aminotransferase (AST): aspartate Aminotransferase (AST) is distributed mainly in the cardiac muscle, and secondarily in the tissues such as the liver, skeletal muscle and kidney. When the serum AST content is lower in normal state, but when the corresponding cells are damaged, the cell membrane permeability is increased, and the AST in cytoplasm is released into blood, so that the serum concentration can be increased, and when the glutamic-oxaloacetic transaminase is obviously increased and the glutamic-oxaloacetic transaminase/glutamic-pyruvic transaminase (ALT) is more than 1, the extensive damage of liver parenchyma is suggested, and the prognosis is poor.

Serum alkaline phosphatase (ALP): is an enzyme widely distributed in human liver, sensitively reflects the damage of hepatocyte and the damage degree, and when hepatocyte membrane is damaged or cell is necrotic, the blood concentration of ALP is increased.

Total Bilirubin (TBIL): the total bilirubin value is the sum of direct bilirubin and indirect bilirubin, the liver plays an important role in the metabolism of bilirubin, the three processes of intake, combination and excretion of unbound bilirubin in blood are performed by liver cells, and TBIL (TBIL) increase can occur when the liver has drug-toxicity hepatitis due to the reflection of liver secretion and excretion functions.

Albumin (Alb): albumin is closely related to liver synthesis dysfunction and also an index reflecting liver function.

Liver function index before and after treatment (see FIG. 12 and FIG. 13)

After 4 weeks of administration, the liver function 5 indexes are compared in a one-factor analysis of variance group, and the indexes before and after treatment have no statistical difference (P > 0.05).

6.2 renal function 3 criteria and their significance in selection

Urea (UA): as an end product of human protein metabolism. Glomerular filtration, in which 40% is absorbed through the proximal tubule, the absorption of the distal tubule depends on the urine volume and is finally excreted from the urine, and spring welcome depends on the concentration of UA in the blood.

Blood uric acid (UREA): uric acid is the end product of purine metabolism, with 80% of total uric acid in the body produced by nuclear protein catabolism and 20% produced by ingestion of purine-rich food catabolism. 1/3 uric acid is discharged from the intestine, 2/3 from the kidney, and is the final product of purine metabolism in the body, and part of serum uric acid is combined with protein, and most of serum uric acid exists in a free form. Most of serum uric acid is filtered by glomeruli and is reabsorbed by 98-100% of proximal tubules, so that the clearance rate of uric acid is low under normal conditions.

Blood Creatinine (CREA): creatinine is a small molecular substance which can be filtered by glomeruli and is rarely absorbed in renal tubules, and creatinine generated in the body every day is almost completely excreted with urine and is generally not influenced by urine volume, so clinical detection of blood creatinine is one of the main methods commonly used for understanding renal function.

The detection of renal function index aims at investigating the change of glomerular filtration function, renal tubule secretion function and pH value after the compound preparation is used. The results show that the indexes before and after treatment have no statistical difference (P > 0.05). The compound of the invention is safe to the kidney function.

Renal function index before and after treatment (see FIG. 12 and FIG. 13)

6.3 Electrocardiogram

The invention detects the electrocardiogram item of the patient (see figure 14)

Before and after treatment, the sinus rhythm has no obvious change and no statistical difference.

6.4 routine blood

Table 10 effect of the compound of the invention on blood routine:

the invention detects 28 items of the blood routine of the patient (see figures 15 and 16)

The blood routine of patients before and after treatment has no obvious change and no statistical difference.

6.5 urinary routine

The invention detects 26 items of the urine routine of the patient (see figure 17)

Through single-factor variance analysis, each index has no significant difference, and the difference has no statistical significance (p is more than 0.05). No obvious adverse reaction is found in the result.

7 conclusion

Clinical research shows that the traditional Chinese medicine composition has an obvious treatment effect on hyperplasia of mammary glands, the symptoms are relieved or disappeared after treatment, the total effective rate of the traditional Chinese medicine composition on breast pain is 77.6%, the total effective rate of lumps is 48.9%, the total curative effect and cure rate of diseases is 3%, the significant efficiency is 26.7%, and the total effective rate is 86.7%. The results of the conventional comparison of the electrocardiogram and the blood of liver function and kidney function of patients before and after treatment and the research of animal experiments on the visceral index show that the medicine is safe.

Biological effects of the invention: the composition can reduce estrogen level and increase progestogen level in peripheral blood of patients, and reduce breast lump to a certain extent.

In conclusion, the invention changes syndrome differentiation treatment into disease differentiation treatment aiming at the characteristics of breast pain and mass of a patient which are diminished and grown along with the change of emotion; aiming at the nodules of breast formed by disharmony of Chong and conception vessels, the treatment follows the principle of reaching the depressed people and consolidating and dispersing, and the compound obtained by compatibility has obvious effect on treating the female breast hyperplasia caused by liver and kidney deficiency and disharmony of Chong and conception vessels, has no toxic or side effect, high safety and low cost, and has practical popularization and application values.

Claims

1. A traditional Chinese medicine composition for treating hyperplasia of mammary glands is characterized in that: the traditional Chinese medicine is prepared from the following raw materials in parts by weight:

2. The traditional Chinese medicine composition according to claim 1, characterized in that: the traditional Chinese medicine is prepared from the following raw materials in parts by weight:

3. The traditional Chinese medicine composition according to claim 2, characterized in that: the traditional Chinese medicine is prepared from the following raw materials in parts by weight:

4. The Chinese medicinal composition according to any one of claims 1 to 3, wherein: the tangerine seeds are salt tangerine seeds.

5. The Chinese medicinal composition according to any one of claims 1 to 3, wherein: the preparation is prepared by taking medicinal powder of raw material medicines, or water or organic solvent extracts of the raw material medicines as active ingredients and adding pharmaceutically acceptable auxiliary materials.

6. The traditional Chinese medicine composition according to claim 5, wherein: the preparation is an oral preparation, and the oral preparation is granules, paste, powder, pills or solution.

7. The preparation method of the traditional Chinese medicine composition as claimed in any one of claims 1 to 6, which is characterized by comprising the following steps: it comprises the following steps:

(1) weighing the raw materials according to the proportion of claim 1;

(2) grinding the raw materials into powder, or extracting with water or organic solvent, and adding adjuvants or auxiliary components.

8. Use of the Chinese medicinal composition of any one of claims 1 to 5 in the preparation of a medicament for treating hyperplasia of mammary glands.