CN114377076A

CN114377076A - Application of traditional Chinese medicine composition in preparation of medicine for preventing and treating hyperplasia of mammary glands

Info

Publication number: CN114377076A
Application number: CN202210117918.9A
Authority: CN
Inventors: 李振宇; 宋洪运; 肖贺; 刑乃栋
Original assignee: Shandong New Time Pharmaceutical Co Ltd
Current assignee: Shandong New Time Pharmaceutical Co Ltd
Priority date: 2021-02-09
Filing date: 2022-02-08
Publication date: 2022-04-22
Also published as: CN112807376A

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition and application thereof. The traditional Chinese medicine composition is mainly prepared from ginseng, rehmannia root, prepared rehmannia root, dogwood fruit, Chinese yam, moutan bark, alisma orientale, tuckahoe, asparagus and dwarf lilyturf tuber, and the invention is a new application developed for ginseng root-strengthening oral liquid which is a product on the market. Pharmacological experiments show that: the traditional Chinese medicine composition can obviously improve rat mammary gland hyperplasia tissue structures caused by exogenous estrogen and P, reduce the content of serum E2 and the positive expression integral of mammary gland tissues ER and PR, and increase the content of serum P.

Description

Application of traditional Chinese medicine composition in preparation of medicine for preventing and treating hyperplasia of mammary glands

Technical Field

The invention relates to a new application of a traditional Chinese medicine composition, in particular to an application of the traditional Chinese medicine composition in preparation of preventing and treating hyperplasia of mammary glands, and belongs to the technical field of traditional Chinese medicines.

Background

Cyclomastopathy refers to hyperplasia of mammary epithelium and fibrous tissue, degenerative changes of mammary tissue ducts and lobules in structure and growth of progressive connective tissue, and the pathogenesis is mainly due to endocrine hormone imbalance. Hyperplasia of mammary glands is the most common breast disease in women, and the incidence rate accounts for the first place of breast diseases.

The hyperplasia of mammary glands belongs to the category of mammary glands in the traditional Chinese medicine, and the mammary glands are considered to be formed by emotional internal injury, disharmony of Chong and conception vessels and phlegm stasis and condensation in the traditional Chinese medicine. Depression, qi stagnation, qi and blood loss, accumulation in the stomach and breast collaterals, obstruction of the channels and collaterals of the breast, and pain caused by obstruction of the channels and collaterals; liver qi transverse adversely affecting the stomach, spleen failing to transport and transform, phlegm turbidity produced internally, qi stagnation and blood stasis with phlegm accumulation as the core, and retained in milk along the meridians, so the milk is caked. The deficiency of liver and kidney and the disorder of Chong and ren meridians are also the important causes of mammary nodules. Kidneys are the root of the five zang organs, and kidneys generate essence to generate sex-stimulating essence, which stimulates Chong and ren meridians, and the Chong and ren meridians descend to the uterus, connect the breasts, and circulate qi and blood, with the milk going upwards and the meridian going downwards. For kidney-qi deficiency, disharmony of thoroughfare and conception vessels, qi and blood stagnation, accumulation of blood stasis in breast and uterus, or breast pain and agglomeration, or disorder of menstruation.

Because the mechanism and the cause of hyperplasia of mammary glands are not exactly known, the current treatment is basically symptomatic treatment. Common treatment methods include massage physiotherapy, hormone therapy, and the like. Or selecting Chinese patent medicine or prescription for treatment under the guidance of syndrome differentiation of traditional Chinese medicine.

The ginseng root-strengthening oral liquid is prepared from ginseng, rehmannia root, prepared rehmannia root, dogwood fruit, Chinese yam, tree peony bark, oriental waterplantain rhizome, poria cocos, radix asparagi and dwarf lilyturf tuber, can be used for treating yin deficiency and qi weakness, consumptive disease and cough, palpitation and short breath, bone steaming and tidal fever, waist soreness and tinnitus and dry stool, but finds a new indication in the actual clinical application process, and provides support for further development and utilization of the product.

Disclosure of Invention

The invention is further developed on the basis of the clinical use feedback of the traditional Chinese patent medicine product ginseng root-strengthening oral liquid, and the ginseng root-strengthening oral liquid has a new application in treating hyperplasia of mammary glands.

The invention aims to provide a new Chinese patent medicine product for treating hyperplasia of mammary glands, and the medicine selectivity of patients is increased. The invention aims to provide the application of the traditional Chinese medicine composition mainly comprising ginseng and the like in preparing the medicine for treating hyperplasia of mammary glands.

The Chinese medicinal composition is prepared from ginseng, rehmannia root, prepared rehmannia root, dogwood fruit, Chinese yam, tree peony bark, oriental waterplantain rhizome, tuckahoe, asparagus and dwarf lilyturf tuber.

Preferably, the ginseng root-strengthening traditional Chinese medicine composition is prepared from the following medicinal materials in parts by weight: 60-90 parts of ginseng, 170 parts of rehmannia root 130-.

The hyperplasia of mammary glands (cyclomastopathy) refers to hyperplasia of mammary epithelial and fibrous tissues, degenerative change of structures of mammary tissue ducts and mammary lobules and growth of progressive connective tissues, and the pathogenesis of the hyperplasia of mammary glands is mainly caused by endocrine hormone imbalance. Hyperplasia of mammary glands is the most common breast disease in women, and the incidence rate accounts for the first place of breast diseases.

According to different types of hyperplasia of mammary glands, the hyperplasia of mammary glands is classified into mastodynia, namely simple hyperplasia of mammary glands, the mastopathy exists that the pathological change is based on the fact that mammary lobules and ducts in breasts are expanded and tissues around the glands are hyperplastic, hyperplasia of epithelial cells of the ducts is taken as a main pathological change, and lumps appearing in the breasts are often cystic hyperplasia with diffuse thickening.

According to the abnormality of the mammary gland structure in quantity and form, the hyperplasia of mammary glands is divided into hyperplasia of mammary tissue, mastopathy (also divided into lobular hyperplasia, fibroglandular disease and fibrosis) and galactocele.

According to the basic tissue changes of hyperplasia of mammary glands, hyperplasia of mammary glands is divided into lobular hyperplasia, fibrosis, inflammation, cyst, epithelial hyperplasia and adenosis.

The invention provides a Chinese patent medicine product for treating hyperplasia of mammary glands.

Preferably, the hyperplasia of mammary glands is selected from the symptoms of mastalgia, namely simple hyperplasia of mammary glands, mastopathy based on the expansion of lobules and ducts of mammary glands in breasts and hyperplasia of tissues around glands exists, hyperplasia of epithelial cells of the ducts of the mammary glands is taken as a main lesion, and mass in the breasts is generally diffuse thickened cystic hyperplasia.

Preferably, the hyperplasia of mammary glands is selected from hyperplasia of mammary glands, mastopathy (also classified into lobular hyperplasia stage, fibroglandular stage and fibrosis stage), and galactocele.

Preferably, the hyperplasia of mammary glands is selected from lobular hyperplasia, fibrosis, inflammation, cyst, epithelial hyperplasia, adenosis.

The second purpose of the invention is to provide a traditional Chinese medicine preparation containing the traditional Chinese medicine composition, namely the traditional Chinese medicine composition is prepared into a clinically acceptable preparation form directly by a conventional process or after pharmaceutically acceptable auxiliary materials are added.

The clinically acceptable dosage form is one or more of capsules, tablets, granules, pills and oral liquid;

preferably, the clinically acceptable dosage form is oral liquid;

further preferably, the oral liquid is ginseng root-strengthening oral liquid.

Compared with the prior art, the invention has the following remarkable technical effects:

the traditional Chinese medicine composition can obviously improve rat mammary gland hyperplasia tissue structures caused by exogenous estrogen and P, reduce the content of serum E2 and the positive expression integral of mammary gland tissues ER and PR, and increase the content of serum P.

Drawings

FIG. 1 is a pathological morphology map (HE staining X200) of the breast tissues of each group in example 1;

FIG. 2 shows ER positive sections of rat mammary tissue (immunohistochemistry × 400);

FIG. 3 shows positive expression staining of PR in rat mammary tissue (immunohistochemistry × 400).

Detailed Description

The inventor explains that the application of the invention is derived from clinical application feedback of the ginseng root-strengthening oral liquid. In the clinical use process of the ginseng root-strengthening oral liquid, the ginseng root-strengthening oral liquid is unexpectedly found to be capable of adjusting female endocrine to a certain extent, reducing breast lumps, improving clinical symptoms of breast distending pain and the like of a patient for individual patients with hyperplasia of mammary glands.

Based on the clinical use feedback of the ginseng root-strengthening oral liquid, the inventor carries out a series of developments on the application of the ginseng root-strengthening oral liquid in the treatment of the hyperplasia of mammary glands, in order to verify the efficacy of the ginseng root-strengthening oral liquid in the treatment of the hyperplasia of mammary glands, the inventor carries out animal test research, only a part of experimental models are taken as examples for description below, and pharmacological experimental research is carried out on other hyperplasia of mammary glands shown in the specification, so that the composition can achieve the same or similar effect, and the effects are not described any more.

The inventor indicates that the following experimental studies are all carried out on the basis of acute toxicity tests and long-term toxicity tests to prove the safety of the drug, and the administration dose in the experimental studies is within a safe dose range.

Effect embodiment of the invention effects of the Chinese medicinal composition on mammary gland hyperplasia model rats

1 Material

1.1 animals

66 female non-pregnant SD rats aged 80-90 days, body mass (200 +/-20) g, license number of experimental animal: SYXK (Shandong) 20180008, provided by New drug Pharmacology center of Lunan pharmaceutical group, Inc., was acclimatized for one week prior to the experiment.

1.2 drugs, reagents

1.2.1 medicaments

The ginseng root-strengthening oral liquid sold in the market is produced by Lunan Qianpu pharmaceutical Co.

Rukuaixiao oral liquid, Guizhou Sanrentang pharmaceutical industry Co.

Estradiol benzoate injection, Hangzhou animal medicine factory.

Progesterone injection, manufactured by Zhejiang Xianju pharmaceutical Co., Ltd.

1.2.2 dosage for rats

Ginseng root strengthening oral liquid: 3.6ml/kg high dose, 1.8ml/kg medium dose, 0.9ml/kg low dose;

oral liquid for treating breast lumps: 1.8 ml/kg.

1.3 Primary reagents

Hematoxylin, eosin, estradiol (E2), progestin (P), ELISA kit and primary antibody, warrior, bosch, bioengineering, ltd.

2. Grouping, modeling and dosing

2.1 grouping

After the animals are fed for 1 week adaptively, the animals are divided into a blank control group, a model control group, a positive control group and ginseng low, medium and high dose groups according to a physical quality random block stratified sampling method, and 11 animals are fed in each group.

2.2 Molding

Reference [1, 2] methods for molding. The hind limb of the rat was injected intramuscularly with estradiol benzoate 0.5mg/(kg · s), 1/d for 20 consecutive days, followed by progesterone 5mg/(kg · s), 1/d, for 5 consecutive days. The hind limb of the rats in the blank control group is injected with the same amount of 0.9% NaCl solution by the same method for 1 time/day and continuously for 25 days. And (3) the successful standard of molding: randomly selecting 3 from the three types of the raw materials, carrying out routine pathological section examination, and determining that the molding is successful if the number of lobules of the mammary gland is increased, the number of lobules of the lobules is increased, the lobules of the lobules are dense and the number of the lobules of the mammary gland is increased, the lobules of the mammary gland are diffused and proliferated, the acinus cavity of the mammary gland is obviously expanded, the ductal epithelium is obviously proliferated, and the three degrees are consistent.

2.3 intervention

The positive control group and each dosage group of ginseng for consolidating constitution are applied with corresponding treatment measures for 30 days from the 1 st day after the successful model building.

The low, medium and high dose groups of the ginseng are respectively administrated with 0.9, 1.8 and 3.6mL/kg according to the conversion of the clinical dose of the animals and the human.

The positive control group is administrated with 1.8ml/kg according to the conversion of the clinical dosage of animals and human beings.

The blank control group and the model control group were drenched with 1.8ml/kg distilled water, and all animals were drenched with 1 time/day.

2.4 specimen Collection and detection

After the last treatment, fasting was 24 h. Rats with 8% sodium sulfide had hair removed around their breasts. Collecting blood from abdominal aorta, centrifuging at 2000r/min for 15min, collecting serum, storing in refrigerator at-70 deg.C, and detecting E2 and P content in the same batch according to ELISA kit specification. The left breast was completely removed and the 2 nd pair was fixed in 10% formaldehyde solution, embedded in paraffin, sectioned (4 μm thick), HE stained, immunohistochemically examined for expression of Estrogen Receptor (ER) and Progesterone Receptor (PR) in rat mammary tissue and positive scores were calculated.

Positive judgment standard reference [3] method and improvement. The staining of the cells was observed under a mirror, and the extent and degree of staining were evaluated. Calculating 200 cells under a high-power visual field (x 400), counting the percentage of positive cells, and respectively counting 1, 2, 3 and 4 points for less than or equal to 25%, 26-50%, 51-75% and > 75%; the staining intensity is counted by 0, 1, 2 and 3 points according to the positive cells with no staining, weak staining (light yellow), medium staining (brown yellow) and strong staining (brown). Each section was randomly scored for 5 fields (x 400), and each field was scored for percent positive cells and staining intensity, which were added as positive expression scores.

2.5 statistical methods

Analysis was performed using SPSS19.0 statistical software. Experimental data on

After the test of the variance, the variance analysis is adopted for the comparison among groups, and the LSD method is adopted for pairwise comparison. P<0.05 indicates that the difference is statistically significant.

3. Examination item

3.1 morphological changes in rat mammary hyperplasia tissue

The left breast was completely dissected and removed 2 nd pair, fixed in 10% formaldehyde solution, paraffin embedded, sectioned (4 μm thick), HE stained.

Mammary gland hyperplasia scoring table	Wu (0)	Slight (1)	Moderate (2)	Severe (3)
					Degree of hyperplasia of mammary glands	0	1	2	3
Number of acini	0	1	2	3
					Degree of dilation of mammary duct	0	1	2	3

3.2 measurement of serum estradiol and Progestagen content in model rat

Collecting blood from abdominal aorta, centrifuging at 2000r/min for 15min, collecting serum, storing in refrigerator at-70 deg.C, and detecting E2 and P content in the same batch according to ELISA kit specification.

3.3 model rat Estrogen receptor and Progestagen receptor Positive expression Observation scores

After each measurement is finished, the expression conditions of Estrogen Receptor (ER) and Progestogen Receptor (PR) of mammary tissue of a rat are detected by immunohistochemistry, and positive points of the expression conditions are calculated. The staining of the cells was observed under a mirror, and the extent and degree of staining were evaluated.

4. Results and conclusions

4.1 morphological changes in mammary gland hyperplasia tissue in groups of rats

The blank control group has no hyperplasia of mammary lobules, a small number of acini, most glands in a static state, dense arrangement and uniform size of glandular epithelial cells, and no obvious secretion in the cavity; the mammary lobules of the model control group are obviously hyperplastic, most acini and partial ducts are in an expanded or extremely expanded state, and are full of secretion, loose in arrangement and obvious in size change; the pathological changes of each treatment group are improved compared with the control group of the model, mammary lobules can be seen to be hyperplastic, compared with the control group of the model, the number of acinus is obviously reduced, duct expansion is obviously relieved, a small amount of secretion and cast-off cells are in the cavity, the arrangement is compact, and the change of the cell size is not obvious.

The results are shown in FIG. 1, Table 1.

Compared with the blank control group rats, the pathological integral of the model control group rats is obviously increased (P < 0.01); the pathological score of rats in each administration group is obviously reduced compared with that of a model control group (P < 0.01); compared with the positive drug, the pathological score of the rats in the high-dose administration group is remarkably reduced (P <0.05), and the pathological score of the rats in the medium-low dose administration group is not remarkably different (P > 0.05).

TABLE 1 morphological changes of mammary gland hyperplasia tissue in groups of rats

Note: compared with the blank control group, the composition of the composition,^#P<0.05，^##P<0.01；

p <0.05, P <0.01, compared to model control;

compared with the positive control group, the delta P is less than 0.05, and the delta P is less than 0.01.

4.2 Effect of different drug interventions on serum estradiol and progestogen levels in model rats

Compared with a blank control group, the E2 level of the model control group is obviously increased, and the P level is obviously reduced (P < 0.01); compared with the model control group, the E2 level of each treatment group is obviously reduced (P <0.05), the P level of the positive control group is obviously increased (P <0.01), and the P level of each dose group of the ginseng consolidating the constitution is increased; compared with the positive group, the level of E2 in the ginseng root-consolidating high-dose group is remarkably increased (P <0.05), and the level of P in each ginseng root-consolidating dose group is remarkably reduced (P < 0.05); compared with the traditional Chinese medicine dosage groups, the ginseng root-strengthening low-dosage group has the best curative effect. The results are shown in Table 2.

TABLE 2 serum estrogen and progestin assay in rats of various groups

p <0.05, P <0.01, compared to model control;

4.3 Effect of different drug intervention on Positive expression of estrogen receptor and Progestagen receptor in model rats

Compared with the blank group, the expression scores of ER and PR in the model group are obviously increased (P < 0.01); compared with the model group, the ER and PR expression scores of each treatment group are remarkably reduced (P is less than 0.05); compared with the positive group, ER of the ginseng root-consolidating high-dose group is remarkably increased (P is less than 0.05), and PR of each ginseng root-consolidating dose group is increased; the ginseng root-consolidating low-dose groups have the best curative effect by comparing the ginseng root-consolidating low-dose groups, and the curative effect is shown in a table 3. Rat mammary tissue ER, PR are nuclear or plasma expressed, and glandular ducts and lobular epithelial nuclei and or cytoplasm are positive when stained. Compared with the blank group, the rat mammary tissue ER and PR in the model group have wide staining range, deeper coloring and obviously enhanced expression, the expression of the ER and PR in the rat mammary tissue of each dose group of the positive group and the ginseng solid is reduced compared with that of the model group, and the expression of the ER and PR in the rat mammary tissue of each dose group of the positive group and the ginseng solid is closer to that of the blank group. The results are shown in Table 3, FIG. 2 and FIG. 3.

TABLE 3 Positive expression of estrogen receptor and progestin receptor in groups of rats

p <0.05, P <0.01, compared to model control;

The traditional Chinese medicine composition can increase the content of serum estrogen, reduce the content of serum progestogen and inhibit the expression of estrogen receptors and progestogen receptors. The experimental results show that the traditional Chinese medicine composition can achieve the purpose of treating hyperplasia of mammary glands.

Reference documents:

[1] guo Xinrong, Zhang Weihua, plum tree orchid, the positive effect on the ovarian pituitary axial hormone regulation in mammary hyperplasia model rats [ J ]. Shandong J. TCM, 2013, 32 (7): 485-486.

[2] Lorentz, junior, huanglanya, etc. homogeneous design methods for the study of mammary gland hyperplasia animal model replication [ J ]. journal of chinese experimental prescriptions, 2011, 17 (7): 204-206.

[3] Chenchen hei, liu bin, brightness. methods for determination of immunohistochemical labeling results [ J ]. journal of china pathology, 2001, 30 (4): 318.

Claims

1. application of a ginseng root-strengthening traditional Chinese medicine composition in preparing a medicine for preventing and treating hyperplasia of mammary glands.

2. The use according to claim 1, wherein the mammary hyperplasia is selected from the group consisting of simple hyperplasia, cystic hyperplasia in which the presence of lesions is based on the enlargement of both lobules and ducts of the mammary glands and on the proliferation of periglandular tissue in the breast, and hyperplasia of epithelial cells in the ducts of the mammary glands is the main lesion and the mass present in the breast is often a diffuse thickening.

3. The use according to claim 1, wherein the hyperplasia of mammary glands is selected from the group consisting of hyperplasia of mammary glands, adenosis of mammary glands, cystic hyperplasia of breast.

4. The use according to claim 1, wherein the hyperplasia of mammary glands is selected from lobular hyperplasia, fibrosis, inflammation, cysts, epithelial hyperplasia, adenosis.

5. The use of claim 1, wherein the ginseng root-strengthening Chinese medicinal composition is prepared from ginseng, rehmannia root, poria cocos, moutan bark, prepared rehmannia root, cornus officinalis, Chinese yam, alisma orientale, asparagus and ophiopogon root.

6. The use of claim 5, wherein the ginseng root-strengthening traditional Chinese medicine composition is prepared from the following medicinal materials in parts by weight: 60-90 parts of ginseng, 170 parts of rehmannia root 130-.

7. The use as claimed in any one of claims 1 to 5, wherein the ginseng root-strengthening traditional Chinese medicine composition does not contain or further contains one or more pharmaceutically acceptable pharmaceutical excipients.

8. The use of claim 7, wherein the ginseng root-strengthening traditional Chinese medicine composition is processed into capsules, tablets, granules, powder, pills or oral liquid by the prior art.

9. The use of claim 8, wherein the ginseng root-securing traditional Chinese medicine composition is a ginseng root-securing oral liquid.