CN110090278B - A medicine containing Gynura procumbens for treating cyclomastopathy, and its preparation method and application - Google Patents

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a medicine for treating hyperplasia of mammary glands (called nodules of breast in traditional Chinese medicine) and a preparation method and application thereof. The medicine is prepared from ethanol extract of Angelica keiskei, water extract of fructus Hordei Germinatus and water extract of fructus crataegi. The compound medicine of the invention can effectively treat and compare the rat with the hyperplasia of mammary glands disease: reducing the content of estradiol in serum and increasing the hormone level of progesterone in the serum; ② the hyperplasia number of lobules in mammary gland tissue is reduced; reducing the number of obliterative and blocked acini in the lobules of the mammary tissue; has good effect of soothing liver and eliminating nodules of breast.

Description

A medicine containing Gynura procumbens for treating cyclomastopathy, and its preparation method and application

Technical Field

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a medicine for treating hyperplasia of mammary glands and a preparation method and application thereof.

Background

Cyclomastopathy is the most common disorder of the breast, occurring in women at any age after the onset of puberty, most commonly in women 30 years-pre-menopause. The incidence rate is up to 60%, and the medicine accounts for the first place of breast diseases. The hyperplasia of mammary glands belongs to benign lesions of mammary glands, refers to normal structural disorder of mammary glands caused by physiological hyperplasia and incomplete involution of lobules of mammary glands in a mature period or periodic change, is not inflammatory and tumor, has obvious breast lumps and severe pain along with menstrual period and emotional change in clinical hyperplasia of mammary glands, has long course of disease, slow progress and easy recurrence, seriously affects the normal life of women, particularly has heavier hyperplasia degree, and can possibly endanger life when the hyperplasia of mammary glands progresses to precancerous lesion stages such as severe hyperplasia of mammary glands, atypical hyperplasia of mammary glands and the like. Therefore, the medicine has important clinical significance for timely curing the patients with the hyperplasia of mammary glands.

Modern medicine considers that hyperplasia of mammary glands is related to endocrine dyscrasia and mental factors. The pathogenesis of the breast cancer is quite complex and is not completely clarified so far, but the common recognition is that the secretion of progestogen and androgen in a patient is insufficient, estrogen is relatively increased, breast tissue is excessively stimulated for a long time, the abstinence and protection effects of the progestogen and the like are lacked, the abnormal rise of prolactin is caused, the normal rhythmical change of the breast tissue is further influenced, the proliferation is excessive, the involution is disordered, the epithelium and fibrous tissue of a duct of the breast and/or the peripheral duct of the gland and the acinus are caused to form cysts, and finally, the hyperplasia of the breast is formed.

At present, the western medicine has no ideal scheme aiming at the clinical treatment of the hyperplasia of mammary glands and lacks specific drugs. Although estrogen receptor antagonists tamoxifen and medicines such as progesterone, methyltestosterone, danazol and the like can be selected, the estrogen receptor antagonists can interfere the balance of human hormones, and the estrogen receptor antagonists are easy to relapse after stopping administration; if the drugs are taken for a long time, adverse reactions such as retinopathy, hepatocellular carcinoma, endometrial cancer and the like are induced, and the long-term acceptance of the drugs by patients is more difficult.

The traditional Chinese medicine recognizes hyperplasia of mammary glands as 'mammary nodules', and the occurrence of the hyperplasia of mammary glands is closely related to the kidney, Chong and ren channels and also has important relation with the liver. The most detailed treatment of this disease is explained by the high grasp of Qing Dynasty, which is pointed out in the book "Yanke Heart Collection, Breast nodule-resolving Breast phlegm-mammary rock treatise": "mammary nodules" well-developed due to depression of liver qi; if it is phlegm-qi stagnation, it is not. Fu Ru belongs to yangming, has a nucleus in milk, and is responsible for liver without yangming? The yangming stomach-qi is most afraid of liver wood, the liver qi is uncomfortable, the stomach shows wood depression, but fears, the stomach is prone to be depressed and cannot be raised, the qi dares to be relaxed, the liver qi is uncomfortable, the swelling is hard to form, the stomach qi dares to be relaxed, the fear is obvious, the pain is not red, and the fear is seen. Not the stomach but the liver but the swelling can be eliminated . Factors related to pathogenesis of breast nodules (hyperplasia of mammary glands) mainly include liver depression, spleen deficiency, kidney deficiency, Chong and ren channel disorder and the like, wherein the deficiency of kidney qi can also cause liver malnutrition, and the dysfunction of liver in smoothing flow causes liver qi stagnation, involvement of regulation of Chong and ren channels and qi and blood, so that breast collaterals are nourished without deficiency, pathogenic factors can affect liver channels, blood stasis is stagnated, phlegm lumps are accumulated, and the breast nodules are formed. So far, although many doctors have deeply and widely dealt with the causes and symptoms of breast nodules from different angles and levels, the disease is considered to have pathological changes of qi stagnation, blood stasis and stagnation leading to the formation of the nodules. Based on these recognitions, Chinese medicine has clinically appeared for treating nodules of breast, such as Xiaojin pills, nodules of breast, Xiaoyao granules, Rukang tablets, etc.; although the Chinese patent medicine has short-term curative effect, the Chinese patent medicine is not suitable for long-term administration of patients because the Chinese patent medicine contains prepared kusnezoff monkshood root, glabrous greenbrier rhizome, Chinese thorowax root or common selfheal fruit-spike and other medicines with obvious toxicity to liver injury, and the repeated and further worsened illness state is easily caused by the non-use of the medicines.

Therefore, the invention aims to search a good product which can be prepared into a prescription for treating the hyperplasia of mammary glands from the plants with food and medicine homology, and achieves the effects of relieving the emotion of a patient, being safe and effective and fundamentally solving the problems during the long-term use of the patient.

Disclosure of Invention

Aiming at the defects in the prior art, the first purpose of the invention is to provide a compound medicine for effectively treating the hyperplasia of mammary glands.

The second purpose of the invention is to provide a preparation method of the compound medicine for treating the hyperplasia of mammary glands.

The third purpose of the invention is to provide the medical application of the compound medicine for treating the hyperplasia of mammary glands.

The invention concept of the invention is as follows:

ancient books record that the breast tuberculosis is hard, small people such as plum and big people such as plum are not moved when pressed, the breast is not moved when pushed, the breast is dull and painful in time, the skin color is as normal, and the qi stagnation of the liver and spleen meridians is formed … …. The meridians and collaterals of the breasts of patients with mild illness are blocked, and pain is caused by obstruction; patients with severe disease may have phlegm coagulation, blood stasis and nodules of breast. I.e., poor emotional depression may easily contribute to the occurrence and development of cyclomastopathy. Therefore, the main therapeutic principles of treating hyperplasia of mammary glands include soothing liver, regulating qi, invigorating stomach, eliminating phlegm, promoting blood circulation and removing blood stasis.

In addition, the inventor of the present application has been working on clinical treatment of liver injury and research work on plant active ingredients for many years, and found that the "food and drug homologous" product angelica keiskei (also known as Gynura procumbens, dai drug paleaction) has good treatment effect on inflammatory pain and liver injury, and the specific mechanism is that (Anti-inflammation and Anti-inflammatory effects are of active reactions in the inflammatory oil from Gynura procumbens, a digital medicine and a new and porous food material. journal of ecological pharmacology 2019,239:111916, doi. org/10.1016/j. jep.2019.111916); ② the medicine can accelerate lipid metabolism in liver cells, eliminate fatty liver, reduce liver injury reaction caused by toxic nitrosamine, and promote the repair of damaged liver tissue. Has the functions of protecting liver, regulating liver metabolism and the like (Gynura procumbens reverts acid and cholesterol-induced liver disease MAPK/SREBP-1c-dependent and-independent pathways. journal of Agricultural and Food Chemistry 2015,63, 8460-dependent 8471; application of Gynura procumbens extract in preparing preparations for preventing and treating liver injury; national patent of invention No. ZL 201310547821.2). The classical medical books record that malt has the effects of promoting qi circulation, promoting digestion, strengthening spleen, stimulating appetite, promoting lactation and relieving distension; haw has the functions of promoting digestion, promoting blood circulation and removing blood stasis. Based on long-term exploration and accumulation, the inventor screens angelica keiskei, malt, hawthorn and the like from products of homology of food and medicine to carry out scientific prescription, and finds that the compound medicine prepared from angelica keiskei, malt, hawthorn and the like has better effects of regulating liver, invigorating stomach, dredging channels, promoting blood circulation and the like, can obviously improve mood, relieve fatigue and the like, but does not know whether the medicine can effectively treat hyperplasia of mammary glands.

Following the therapeutic principle of 'dredging, regulating and tonifying' adopted by the traditional Chinese medicine for treating nodules of breast (called as hyperplasia of mammary glands in Western medicine), the inventor carries out a great deal of scientific experimental research on whether the compound medicine has the curative effect of treating the hyperplasia of mammary glands, and researches the improvement effect of the medicine administration of the invention on corresponding lesions by establishing an animal model of the hyperplasia of mammary glands. The research result shows that the compound medicine can: reducing the height of the nipple and the diameter of the nipple; ② the increase of estradiol in serum is reduced and the hormone level of progesterone in serum is increased; (iii) reducing the number of lobules and obliterating acini in the mammary tissue. Therefore, the traditional Chinese medicine has the efficacy of soothing liver and eliminating mammary nodules.

In order to achieve the first object, the invention adopts the following technical measures:

a medicine for treating hyperplasia of mammary glands is prepared from ethanol extract of Angelica keiskei, water extract of fructus Hordei Germinatus and water extract of fructus crataegi;

further, the medicine for treating the hyperplasia of mammary glands is prepared from an ethanol extract of angelica keiskei, a water extract of malt and a water extract of hawthorn;

further, the medicine for treating the hyperplasia of mammary glands is prepared from an ethanol extract of angelica keiskei, a water extract of malt, a water extract of hawthorn and auxiliary materials;

further, the auxiliary materials are starch and dextrin;

furthermore, the auxiliary materials are starch: dextrin 4: 1, starch, dextrin mixtures;

further, the preparation method of the angelica keiskei ethanol extract comprises the following steps: pulverizing dry stems of Angelica keiskei, sieving (sieving with eighty mesh sieve), adding 70-90 v/v% ethanol, reflux extracting at 70-90 deg.C for 1-3 times, filtering, mixing filtrates, concentrating, and drying to obtain Angelica keiskei ethanol extract.

Further, the reflux extraction process: the time of each reflux extraction is 2h, and the ratio of the ethanol addition amount of 70-90 v/v% of the Angelica keiskei to the dry stem amount of the Angelica keiskei is 1L (0.05-0.5) kg, preferably 1L (0.05-0.2) kg, and optimally 1L:0.1 kg.

Preferably, the preparation method of the angelica keiskei ethanol extract comprises the following steps: pulverizing dry stems of Angelica keiskei, sieving (sieving with eighty mesh sieve), adding 80 v/v% ethanol, reflux-extracting at 85 deg.C for 3 times (2 hr each time, the ratio of 80 v/v% ethanol to dry stems of Angelica keiskei is 1L:0.1kg), filtering, mixing filtrates, concentrating, and drying to obtain Angelica keiskei ethanol extract.

Further, the concentration and drying process is to use a rotary evaporator to concentrate and dry at 60 ℃ to obtain dry extract, namely the angelica keiskei ethanol extract.

Further, the preparation method of the malt water extract comprises the following steps: drying, pulverizing and sieving fructus Hordei Germinatus (raw fructus Hordei Germinatus) with eighty mesh sieve, adding water, reflux extracting at 50-80 deg.C for 1-3 times, filtering, mixing filtrates, concentrating, and drying to obtain fructus Hordei Germinatus water extract.

Further, the reflux extraction process: the time of each reflux extraction is 2h, and the ratio of the addition amount of water to the amount of raw malt after drying is 1L (0.05-0.5) kg, preferably 1L (0.05-0.2) kg, and most preferably 1L (0.1 kg).

Preferably, the preparation method of the malt water extract comprises the following steps: drying raw fructus Hordei Germinatus, pulverizing, sieving (sieving with eighty mesh sieve), adding water, reflux-extracting at 60 deg.C for 3 times (2 hr for each time, the ratio of water added in reflux-extracting fructus Hordei Germinatus to dried fructus Hordei Germinatus is 1L:0.1kg), filtering, mixing filtrates, concentrating, and drying to obtain fructus Hordei Germinatus water extract.

Further, the concentration and drying process is to use a rotary evaporator to concentrate and dry at 60 ℃ to obtain dry extract, namely the malt water extract.

Further, the preparation method of the hawthorn water extract comprises the following steps: drying fructus crataegi, pulverizing, sieving (sieving with eighty mesh sieve), adding water, reflux extracting at 50-80 deg.C for 2-4 times, filtering, mixing filtrates, concentrating, and drying to obtain fructus crataegi water extract.

Further, the reflux extraction process: the time of each reflux extraction is 2h, and the ratio of the addition amount of water to the amount of dried fructus crataegi is 1L (0.05-0.5) kg, preferably 1L (0.05-0.2) kg, and most preferably 1L (0.1 kg).

Preferably, the preparation method of the hawthorn water extract comprises the following steps: drying fructus crataegi, pulverizing, sieving (sieving with eighty mesh sieve), adding water, reflux-extracting at 60 deg.C for 3 times (2 hr for each time, the ratio of water added in reflux-extracting fructus crataegi to dried fructus crataegi is 1L:0.1kg), filtering, mixing filtrates, concentrating, and drying to obtain fructus crataegi water extract.

Further, the concentration and drying process is to use a rotary evaporator to concentrate and dry at 60 ℃ to obtain dry extract, namely the hawthorn water extract.

Further, the medicine for treating the hyperplasia of mammary glands is prepared from the raw materials of 400-1000 parts by weight of angelica keiskei ethanol extract, 100 parts by weight of malt water extract and 80-200 parts by weight of hawthorn water extract.

Furthermore, the medicine for treating the hyperplasia of mammary glands is prepared from the raw materials comprising 800 parts by weight of angelica keiskei ethanol extract, 100 parts by weight of malt water extract and 150 parts by weight of hawthorn water extract.

Furthermore, the medicine for treating the hyperplasia of mammary glands is prepared from 800 parts by weight of angelica keiskei ethanol extract, 100 parts by weight of malt water extract, 150 parts by weight of hawthorn water extract and auxiliary materials, wherein the weight of the auxiliary materials is 1-3 times, preferably 1.5-2.5 times, and most preferably 2 times of the total weight of the angelica keiskei ethanol extract, the malt water extract and the hawthorn water extract;

in order to achieve the second purpose, the invention adopts the following technical measures:

a preparation method of a medicine for treating hyperplasia of mammary glands comprises the following steps:

mixing ethanol extract of Angelica keiskei, water extract of fructus Hordei Germinatus and water extract of fructus crataegi at a certain proportion, pulverizing, and sieving (80-200 mesh sieve) to obtain compound medicine.

In order to realize the third purpose, the invention also provides application of the compound medicine consisting of the extract in preparing medicines for treating hyperplasia of mammary glands, in particular to application in preparing medicines for treating lobular hyperplasia and/or obliterative gonad.

The formula principle of each raw material of the invention is as follows:

angelica keiskei is mild in nature and sweet in taste, and is good at soothing liver, relieving depression, promoting qi circulation and removing food stagnation, and is used as a monarch drug; malt, neutral in nature and sweet in taste, has the effects of invigorating spleen and stomach channels, is good at promoting qi circulation, promoting digestion, invigorating spleen, benefiting stomach, promoting lactation and relieving flatulence, and is used as a minister; haw, fructus crataegi, sour and sweet, slightly warm, can enter stomach to eliminate indigestion, subdue liver to dissipate blood stasis, promote qi circulation to relieve pain, and has the efficacy of softening liver, resolving depression and eliminating nodules of breast as adjuvant and guiding the synergistic medicines.

Compared with the prior art, the invention has the following advantages and effects:

1. the raw materials required by the pharmaceutical compound preparation are all products with food and medicine homology, are natural, have no toxicity of organs (particularly liver), and are more easily accepted by patients and friends; the whole extraction method and the preparation process are simple and easy to implement.

2. The medicine prepared by the invention can effectively treat hyperplasia of mammary glands, in particular to the hypotype of hyperplasia of mammary glands, such as lobular hyperplasia and obliterative gonad. The expression can eliminate the level abnormality of estradiol and progesterone in blood serum, reduce the lobular hyperplasia number and the obliterated glandular tube number of mammary glands and maintain the normal tissue morphology of the mammary glands. Compared with the existing mainstream therapy, the medicine for treating the hyperplasia of mammary glands has the advantages of good effect, low price and the like.

3. The medicine provided by the invention integrates the products for soothing liver, dispersing depressed vital energy, promoting lactation, relieving distension, promoting qi circulation and removing blood stasis, is scientifically combined together, and has the effects of regulating liver, invigorating stomach, dredging channels, promoting blood circulation and the like in a synergistic manner, so that the effect of preventing and treating the hyperplasia of mammary glands by using the medicine provided by the invention is achieved.

Drawings

FIG. 1 is a graph showing the effect of formulation 1 of example 3 on the treatment of abnormal estradiol levels in the serum of mammary gland hyperplasia rats.

FIG. 2 is a graph showing the effect of formulation 1 in example 3 on the treatment of abnormal progesterone levels in serum of mammary gland hyperplasia rats.

FIG. 3 is a graph showing the effect of formulation 1 in example 3 on the treatment of abnormal levels of the four indices of papillary height, diameter, estradiol and progesterone in mammary gland hyperplasia rats.

FIG. 4 is a graph showing the effect of the preparation 2 of example 4 on the treatment of abnormal levels of both lobular hyperplasia of mammary glands and obliterated glandular ducts in rats with cyclomastopathy.

Detailed Description

The applicant shall now describe in further detail the product of the invention, its preparation and use in relation to specific examples. It should be understood that the following should not be taken as limiting the scope of the invention in any way.

The applicant carries out series and large amount of scientific experiments on the effect of the medicine for treating the hyperplasia of mammary glands, and confirms that the compound medicine can effectively recover the levels of estradiol and progesterone in the serum of a model rat. Meanwhile, the mammary gland lobular hyperplasia number and the number of mammary gland obliterating acinus can be effectively reduced, and the morphological structure of mammary gland tissue can be repaired.

Example 1 a process for the preparation of formulation 1 comprising the steps of:

(1) preparation of Angelica keiskei extract

Crushing 5kg of dried whole angelica keiskei into powder which is completely sieved by a 80-mesh sieve, placing the powder into an extraction tank, adding 25L of 80% (v/v) ethanol water solution into the extraction tank each time, carrying out reflux extraction for 2 times and 2 hours each time at 60 ℃, combining the 2 times of extraction solutions, carrying out reduced pressure concentration and freeze drying to obtain 876.5g of angelica keiskei extract powder for later use.

(2) Preparation of malt extract

Pulverizing 3kg dried fructus Hordei Germinatus (raw) into powder completely sieving with 80 mesh sieve, placing into extraction tank, adding 15L water each time, decocting at 60 deg.C for 2 times, each time for 1.5 hr, mixing the 2 extractive solutions, concentrating under reduced pressure, and freeze drying to obtain fructus Hordei Germinatus extract powder 330.0 g.

(3) Preparation of hawthorn extract

Pulverizing 2kg dried fructus crataegi into powder, sieving with 80 mesh sieve, placing into extraction tank, adding 10L water each time, decocting at 60 deg.C for 2 times, each for 1.5 hr, mixing 2 times of extractive solutions, concentrating under reduced pressure, and freeze drying to obtain folium Hordei Germinatus extract powder 235.5 g.

(4) Mixing 500g of angelica keiskei extract powder obtained in step (1), 100g of malt extract powder obtained in step (2) and 150g of hawthorn extract powder obtained in step (3), uniformly mixing, and crushing to pass through a 80-mesh sieve to obtain mixed extract powder; the preparation 1 is bottled and stored in a sealed manner.

Further, a mixture of starch and dextrin with the weight 2 times of that of the mixed extract powder obtained in the step (4) can be added (the weight ratio of the starch to the dextrin in the mixture is 4: 1); during the stirring period after the uniform mixing, adding a 95% (v/v) ethanol aqueous solution with the total amount of 100mL in a spraying mode, uniformly mixing to prepare a soft material, granulating by adopting a compression granulator, and drying in an oven with the temperature set at 60 ℃ for 24 hours in a dish to obtain the granular preparation, and bottling and storing in a sealed manner. The following examples are not repeated.

Example 2 a method of preparing formulation 2 comprising the steps of:

(1) preparation of Angelica keiskei extract

Crushing 5kg of dried whole angelica keiskei into powder which is completely sieved by a 80-mesh sieve, placing the powder into an extraction tank, adding 50L of 80% (v/v) ethanol water solution into the extraction tank each time, carrying out reflux extraction for 2 times at 80 ℃ for 1 hour each time, combining the 2 times of extraction solutions, carrying out reduced pressure concentration and freeze drying to obtain 1035.9g of angelica keiskei extract powder for later use.

(2) Preparation of malt extract

Pulverizing 3kg dried fructus Hordei Germinatus (raw) into powder completely sieving with 80 mesh sieve, placing into extraction tank, adding 30L water each time, decocting at 80 deg.C for 2 times, each time for 1 hr, mixing the 2 extractive solutions, concentrating under reduced pressure, and freeze drying to obtain 381.0g fructus Hordei Germinatus extract powder for use.

(3) Preparation of hawthorn extract

Pulverizing dried fructus crataegi 5kg into powder, sieving with 80 mesh sieve, placing into extraction tank, adding 50L water each time, reflux extracting at 80 deg.C for 2 times, each for 1.5 hr, mixing 2 extractive solutions, concentrating under reduced pressure, and freeze drying to obtain fructus crataegi extract powder 620.6 g.

(4) Mixing 800g of angelica keiskei extract powder obtained in step (1), 100g of malt extract powder obtained in step (2) and 100g of hawthorn extract powder obtained in step (3), uniformly mixing, and crushing to pass through a 80-mesh sieve to obtain mixed extract powder; the preparation 2 is bottled and stored in a sealed way.

EXAMPLE 3 therapeutic Effect of formulation 1 on mammary hyperplasia in rats

Female SD infertile rats (provided by the centers for disease prevention and control in Hubei province) weighing 180 g. The water is freely drunk, the room temperature of the breeding room is kept at 25 +/-1 ℃, and the light and shade period is 12 hours.

(1) Modeling and administration of hyperplasia of mammary glands

During the experiment period, the pellet feed is fed, and the experimental period is entered after 1 week of observation feeding. Female SD rats were randomly divided into 5 groups (5 per group): the breast nodule removing capsule is a capsule prepared from antler, dandelion, kelp, pseudo-ginseng, red paeony root, seaweed, uniflower swisscentaury root, costustoot, figwort root, motherwort herb, suberect spatholobus stem, thin evodia leaf, weeping forsythia capsule, leatherleaf mahonia stem, glabrous greenbrier rhizome and other medicaments, has the effects of promoting blood circulation to remove blood stasis and softening and resolving hard mass, and is clinically commonly used for the breast nodule caused by qi stagnation and blood stasis, and comprises breast lobular hyperplasia, ovarian cyst, uterine fibroid and other symptoms pointed by western medicine. Except for a blank control group, the rats respectively receive intramuscular injection of estradiol benzoate (0.3 mg/kg; Sichuan gold pharmaceutical Co., Ltd.) and progesterone (4 mg/kg; Chifeng Bonn pharmaceutical Co., Ltd.) on two thighs, and the mammary gland hyperplasia model is formed after 1 time every other day and 15 times continuously for 30 days, wherein the physiological saline is injected into the thighs of the rats in the other groups in an amount of 0.2ml/100 g.

After the model is established, rats in the blank group and the model group receive physiological saline (1ml/100g) with the same volume and are perfused with stomach; the rats in the low and high dose groups respectively receive the preparation 1 (the preparation is dissolved by water) of 300 mg/kg/time and 900 mg/kg/time of intragastric administration for 1 time every other day and 15 times in total for 30 consecutive days; the mammary nodule eliminating group rat receives mammary nodule eliminating (the dosage of adult is 5 tablets each time, and is converted into rat dosage of 210mg/kg), and the administration is performed for 30 days, 1 time every other day and 15 times in total.

(2) Height and diameter index detection of the second pair of mammary glands of rat

Carrying out breast periphery depilation on the second pair of breasts of the rat by using depilatory cream (France Weiting Veet), then marking the height by taking the paperboard as a reference, and measuring the paperboard marking height by using a vernier caliper (Shanghai platform sea work volume Co., Ltd); the diameter of the nipple was measured directly with a vernier caliper.

(3) Detection of estradiol and progesterone indexes in rat serum

The method comprises the steps of taking serum of a fully-anesthetized rat, completing detection of the contents of two hormone indexes of estradiol and progesterone in all serum samples by using a multifunctional enzyme-linked immunosorbent assay kit (Shanghai enzyme-linked biotechnology, Inc.) according to a standard program on a specification of the estradiol and progesterone enzyme-linked adsorption assay kit, and researching the influence of the preparation 1 on the internal secretion of the rat with the mastoplasia.

(4) Results of the experiment

The results of measuring the estradiol content in the serum are shown in fig. 1. The method comprises the following specific steps:

estradiol (E)₂) Is a female hormone which exerts its estrogenic effects mainly by entering into the nucleus to bind to the estrogen receptor ER to form a dimer, activating the intracellular estrogenic response. In pathological states, E₂The content is abnormally increased, ER is combined and increased, and the medicine acts on sensitive mammary tissue for a long time and stimulates and induces the hyperproliferation of mammary cells. Therefore, the content of the estradiol is detected in the invention, which is beneficial to assisting the judgment of the disease degree of the hyperplasia of mammary glands.

At the end of the first week of administration, compared with a blank control group, the estradiol index value in the serum of a model group almost reaches three times (P is less than 0.001) of that of the control group, and in an administration intervention group, the low and high doses of the group have a dose-dependent recovery effect on the abnormally high value of the estradiol, wherein the effect of the high dose group is the best (the P value is less than 0.05), but the effect is slightly weaker than that of nodules of breast; denotes P value less than 0.01 compared to the blank control group; and, #, indicates that P values are less than 0.05, 0.01, respectively, compared to the model group.

Secondly, when the fourth administration is finished, the whole difference of the serum estradiol values in each group is reduced (possibly, the body has the effect of regulating the estradiol while the medicine is effectively intervened); the low and high doses of formulation 1 group exhibited dose-dependent recovery from abnormally high values of estradiol, with the high dose group having the best effect (P value less than 0.01), almost equivalent to nodules of breast. Denotes a P value of less than 0.05 compared to the blank control group; and, #, indicates that P values are less than 0.05, 0.01, respectively, compared to the model group.

The results of measuring the content of progesterone in serum are shown in fig. 2. The method comprises the following specific steps:

with the change of the menstrual cyclic hormone level, the mammary tissue structure undergoes the physiological periodic changes of 'hyperplasia' and 'involution'. Wherein, the estradiol is used for promoting the growth of fibrous tissues around the mammary duct and the duct, and the progesterone is used for promoting the development of mammary lobules and acinar tissues. Once the estradiol level in vivo is abnormally increased and the progesterone level is reduced, the mammary tissue stimulated by estradiol for a long time can cause hyperproliferation and involution insufficiency of mammary glands due to the lack of the control and protection effects of progesterone, so that the hyperplasia and the hyperplasia of the mammary tissue structure are disordered. Therefore, the content of the progesterone is detected in the invention, which is beneficial to assisting the repair state judgment of the hyperplasia of mammary glands.

Compared with a blank control group, the index value of the progesterone in the serum of a model group rat is almost reduced to be less than half of that of the control group (P is less than 0.001), and in a dosing intervention group, the low and high doses of a preparation 1 group have a dose-dependent recovery effect on the abnormally low value of the progesterone in the serum of the rat, wherein the improvement effect of the preparation 1 high dose group is the best (the P value is less than 0.01), but the nodules of breast are not obviously eliminated; indicates P value less than 0.001 compared to the blank control group; # indicates a P value of less than 0.01 compared to the model group.

Secondly, when the administration is finished in the fourth week, the whole difference of the progesterone values in the serum of each group of rats is reduced, and the body can also have the effect of adjusting the progesterone while the medicine is effectively intervened (the specific mechanism is not clear).

The results of abnormal heights and diameters of rat papillae and of the levels of the four indicators estradiol and progesterone in serum are shown in fig. 3. The method comprises the following specific steps:

the theory of traditional Chinese medicine considers that the breasts have close relationship with meridians, viscera and qi and blood. The liver meridian of foot jueyin goes through the diaphragm, spreads over the chest and hypochondrium, and runs around the nipple, indicating that the nipple belongs to the liver. Therefore, the applicant of the present invention considers that the height, the diameter, etc. of the nipple are the evaluation indexes of the mammary gland hyperplasia behavior, combines the evaluation indexes with the level change of estradiol and progesterone in the same sample, and then carries out comprehensive evaluation through the contribution values (area scores) of four indexes which are closely related to the mammary gland hyperplasia degree through the radar chart, so that the related multiple indexes can be simultaneously evaluated relatively easily, and the true information of the mammary gland lesion can be reflected more easily.

Note: the radar chart analysis method is a multivariate comparative analysis technology constructed based on a graph similar to a navigation radar display screen. The method can be used for simultaneously carrying out comparative analysis on a plurality of indexes and analyzing the change of the same index in different periods. And the evaluator compares the radar images and gives a qualitative comprehensive evaluation result by observation. The four indexes are the height and diameter of the papilla of the rat and the four items of estradiol and progesterone in serum.

At the end of the first week after administration, compared with a blank control group, the significance of four index contribution values (area points) of a model group is increased (P is less than 0.001), and in an administration intervention group, the low and high doses of the preparation 1 group have dose-dependent inhibition effects on the four index contribution values (the P values are respectively less than 0.05 and 0.01), wherein the effect of the preparation 1 high dose group is basically the same as that of a mammary nodule elimination group; indicates P value less than 0.001 compared to the blank control group; # #, # # ## # indicates that the P values are less than 0.05, 0.01 and 0.001 respectively compared with the model group.

Secondly, at the end of the fourth administration period, the overall difference of the contribution values of the four indexes of each group is reduced (possibly, the body has a callback result while the medicine intervenes effectively), but the four contribution values of the preparation 1 can still be effectively reduced (the P value is less than 0.05), and the curative effect of the high dose is basically equal to that of the nodules of breast. Denotes a P value of less than 0.05 compared to the blank control group; and, #, indicates that P values are less than 0.05, 0.01, respectively, compared to the model group.

EXAMPLE 4 therapeutic Effect of formulation 2 on mammary hyperplasia in rats

Female SD infertile rats (provided by the centers for disease prevention and control in Hubei province) weighing 180 g. The water is freely drunk, the room temperature of the breeding room is kept at 25 +/-1 ℃, and the light and shade period is 12 hours.

(1) Modeling and administration of hyperplasia of mammary glands

During the experiment period, the pellet feed is fed, and the experimental period is entered after 1 week of observation feeding. Female SD rats were randomly divided into 5 groups (5 per group): blank control group, model group, low dose group of formulation 2, high dose group of formulation 2, and nodules of breast. Except for a blank control group which is injected with physiological saline intramuscularly (according to the dosage of 0.2ml/100 g), two thighs of the other groups of rats respectively receive the injection of estradiol benzoate (0.3mg/kg) and progesterone (4mg/kg) intramuscularly for 1 time every other day and 15 times continuously for 30 days to form a mammary gland hyperplasia model.

After the model is established, rats in the blank group and the model group receive physiological saline (1ml/100g) with the same volume and are perfused with stomach; the rats in the low and high dose groups respectively receive preparation 2 (dissolved in water) of 300 mg/kg/time and 900 mg/kg/time of intragastric administration for 1 time every other day and 15 times in total for 30 consecutive days; the mammary nodule eliminating group rat receives mammary nodule eliminating (the dosage of adult is 5 tablets each time, and is converted into rat dosage of 210mg/kg), and the administration is performed for 30 days, 1 time every other day and 15 times in total.

(2) H & E staining assay of rat second pair of mammary tissue

After dosing, the rats were sacrificed under anesthesia, and the second pair of mammary glands of the rats were removed and fixed with 10% formalin, paraffin embedded, sectioned, and finally H & E stained. The mammary gland tissue structure of the rat is observed under a light microscope, and the influence of the preparation 2 on the mammary gland hyperplasia of the rat is researched.

(3) Results of the experiment

Pathological examination is always the gold standard for judging the nature and degree of pathological changes of organs or tissues. Therefore, in addition to the aforementioned estrogen levels and behavioral assessment of the breast, it is highly desirable in the present invention to confirm the specific lesions inherent in breast hyperplasia using H & E staining.

FIG. 4 shows the staining pattern of HE of mammary gland of experimental rat with mammary gland hyperplasia in example 4. Observed from rat mammary tissue sections: (1) the shape, size and number of lobule of mammary gland of the rat of the blank control group are normal, the interstitial tissue is obvious, and the lumen of mammary gland is not expanded; (2) the mammary tissue mesenchyme of the model group rat is not obvious, the number of lobules is obviously increased (solid arrow), the volume is enlarged, and the number of acini is increased; part of the acinus and lumen are in a clearly dilated state and are accompanied by the appearance of a large number of secretions (open arrows) in the lumen, manifested as lobular hyperplasia and obliterative gonadal symptoms; (3) compared with the model group, the rats in the low-dose group of the preparation 2 have reduced mammary lumen dilation degree, and the number of lobules and obliterative acini is obviously reduced; (4) compared with the model group, the number of lobules of mammary gland and closed acinus of the rat in the preparation 2 high-dose group is obviously reduced, and the interstitium is obvious; the improvement effect of the preparation 2 on the hyperplasia of mammary glands with high dose is almost the same as that of the breast nodule eliminating group. Indicates P value less than 0.001 compared to the blank control group; # #, # # ## # indicates that the P values are less than 0.05, 0.01 and 0.001 respectively compared with the model group.

Claims (6)

1. A medicine for treating hyperplasia of mammary glands;

the medicine for treating hyperplasia of mammary glands is prepared from 400-1000 parts by weight of gynura procumbens ethanol extract, 100 parts by weight of malt water extract and 80-200 parts by weight of hawthorn water extract;

the gynura procumbens refers to the whole gynura procumbens;

the ethanol is 70-90 v/v% ethanol.

2. The medicine for treating hyperplasia of mammary glands according to claim 1, wherein the ethanol extract of gynura procumbens is prepared by the following steps: pulverizing dried whole Gynura procumbens, sieving, adding 70-90 v/v% ethanol, reflux extracting at 70-90 deg.C for 1-3 times, filtering, mixing filtrates, concentrating, and drying to obtain Gynura procumbens ethanol extract.

3. The drug for treating mammary gland hyperplasia according to claim 1, wherein the malt water extract is prepared by the following method: drying raw fructus Hordei Germinatus, pulverizing, sieving, adding water, reflux extracting at 50-80 deg.C for 1-3 times, filtering, mixing filtrates, concentrating, and drying to obtain fructus Hordei Germinatus water extract.

4. The medicine for treating hyperplasia of mammary glands according to claim 1, wherein the water extract of hawthorn is prepared by the following steps: drying fructus crataegi, pulverizing, sieving, adding water, reflux extracting at 50-80 deg.C for 2-4 times, filtering, mixing filtrates, concentrating, and drying to obtain fructus crataegi water extract.

5. The drug for treating hyperplasia of mammary glands as claimed in claim 1, wherein the drug for treating hyperplasia of mammary glands is prepared from the raw materials of 500-800 parts by weight of Gynura procumbens ethanol extract, 100 parts by weight of malt water extract and 100-150 parts by weight of hawthorn water extract.

6. Use of a medicament according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of a cyclomastopathy.

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