CN115919939B - Traditional Chinese medicine composition for treating hyperplasia of mammary glands and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition for treating hyperplasia of mammary glands and preparation method and application thereof Download PDFInfo
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- CN115919939B CN115919939B CN202211548831.3A CN202211548831A CN115919939B CN 115919939 B CN115919939 B CN 115919939B CN 202211548831 A CN202211548831 A CN 202211548831A CN 115919939 B CN115919939 B CN 115919939B
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- hyperplasia
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Abstract
The application relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for treating hyperplasia of mammary glands, and a preparation method and application thereof. The application takes the radix ranunculi ternati, the ligusticum wallichii, the radix bupleuri, the blumea balsamifera, the green tangerine peel, the poria cocos and the mirabilite as raw materials and takes the radix ranunculi ternati as the monarch drug. Mainly takes the effects of bupleurum root of soothing liver and relieving depression; blumea balsamifera has pungent and warm nature, is good in warming the middle-jiao and activating blood, detoxifies and relieves pain and swelling, and has good effects of warming the middle-jiao and activating blood, relieving pain and swelling; is used as ministerial drug together with rhizoma Ligustici Chuanxiong for replenishing blood, promoting blood circulation for removing blood stasis, activating qi-flowing, relieving pain, and promoting fragrance. Taking the effects of strengthening spleen, promoting diuresis, removing dampness and resolving phlegm of poria cocos; it is combined with Qing Pi to soothe liver and regulate qi, dispel stasis and eliminate phlegm as adjuvant drug. Natrii sulfas is effective in clearing heat, relieving swelling, softening hardness, and resolving hard mass. The medicines are compatible, and the functions of soothing liver, regulating qi, activating blood, dissolving stasis, softening hardness and resolving hard mass are collected together. Finally, a modern Chinese medicine external preparation with good treatment effect on hyperplasia of mammary glands is obtained.
Description
Technical Field
The application relates to the technical field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for treating hyperplasia of mammary glands, and a preparation method and application thereof.
Background
The hyperplasia of mammary glands is a common non-inflammatory and non-neoplastic disease of women, and has the characteristics of long disease course, slow development, easy recurrence and the like. The incidence rate of women of childbearing age is counted to be more than 60 percent. Along with the acceleration of the pace of social life and the recent environmental pollution and other reasons, the prevalence rate has recently shown a trend of rising year by year, clinical symptoms mainly include breast pain, breast lumps and nipple discharge, economic and mental stress is brought to female patients, physical and mental health of the patients is seriously affected, and social importance is increasingly brought to.
Western medicine considers that the pathogenesis of hyperplasia of mammary glands is related to endocrine dyscrasia and mental factors, in particular hypothalamus-pituitary-ovary-mammary gland endocrine axis imbalance, and ovarian secretion dysfunction leads to reduced lutein secretion, relatively increased estrogen, and finally hyperplasia of mammary epithelium and fibrous tissue, structural degenerative disease of mammary tissue ducts and lobules and progressive connective tissue growth. Most of western medicine adopts hormone, vitamin medicine, iodine agent and other symptomatic treatment medicines, and the medicines can regulate the in-vivo estrogen level and relieve pain, but the phenomena of hectic fever, night sweat, headache, dysphoria, hypomenorrhea, even stop menstruation, unstable curative effect and the like exist after a high proportion of patients take the medicine, so that the medicine is in great dispute and is not preferred.
The traditional Chinese medicine considers that hyperplasia of mammary glands belongs to the scope of 'nodules of breast' in the traditional Chinese medicine, and the pathogenesis of the hyperplasia of mammary glands is: the women use the conception vessel as the basis, and if the women lose the coordination, the thoroughfare vessel is incoordination, or pathogenic wind attack, qi is not dispersed, and the women accumulate in the breast, or are hard or swelling, and pain has a core. The symptoms of hyperplasia of mammary glands are caused by depression of liver-qi, deficiency of liver and kidney, disorder of thoroughfare and conception vessels, phlegm coagulation and blood stasis due to emotional, tiredness, diet, constitution and other reasons. Hyperplasia of the mammary glands is often manifested by pain in the breast and breast touch to the nodules, which are more pronounced and aggravated before menstruation. The hyperplasia of mammary glands has various pathological types, such as no active conditioning and treatment, and brings great pain and pressure to the body and mind of the patient, and serious and possibly worsening canceration. The traditional Chinese medicine considers that the mammary gland diseases are closely related to meridians such as liver, stomach, thoroughfare, conception vessels and the like. The breast caking disease is mainly caused by unsmooth daily life or work, high pressure, excessive anxiety and anger, liver and spleen injury, liver depression and qi stagnation and spleen and stomach disorder, spleen and stomach can not normally transport and transform water dampness, so that phlegm is accumulated in the water dampness, and if phlegm and qi are mutually accumulated in the breast, stasis and agglomeration of breast nodules are formed. Some patients also have yang deficiency and phlegm dampness accumulation due to liver and kidney deficiency and disharmony of thoroughfare and conception vessels.
The existing external traditional Chinese medicine preparation for treating hyperplasia of mammary glands has the problems of rough process, unstable quality, incapability of ensuring consistency and stability of medication, inaccurate medication dosage, larger dosage and inconvenience for popularization and application.
For example, patent application CN201710666693.1 (a traditional Chinese medicine for treating hyperplasia of mammary glands) specifically discloses a traditional Chinese medicine for treating hyperplasia of mammary glands, which is prepared from the following components: 8-12 parts of epimedium herb, 8-12 parts of cistanche salsa, 3-7 parts of cornu cervi degelatinatum, 8-12 parts of radix bupleuri, 8-12 parts of nutgrass galingale rhizome, 8-12 parts of Chinese angelica, 8-12 parts of white paeony root, 8-12 parts of szechuan lovage rhizome, 3-7 parts of bulbus fritillariae cirrhosae, 8-12 parts of common selfheal fruit-spike, 8-12 parts of full melon, 3-7 parts of rhubarb, 3-7 parts of mirabilite, 3-7 parts of myrrh, 8-12 parts of seaweed, 8-12 parts of kelp and 3-7 parts of borneol; processing the above materials, and making into therapeutic patch. The patch prepared by the method has poor absorption effect and needs to be used together with equipment such as a massager.
For example, patent CN201310142141.2 (an external medicine for treating hyperplasia of mammary glands and a preparation method thereof), an external medicine for treating hyperplasia of mammary glands comprises the following components in parts by weight: 2.38 to 34.09 portions of radix ranunculi ternati, 1.79 to 34.09 portions of rhizoma corydalis, 5.35 to 45.46 portions of radix semiaquilegiae, 1.79 to 40.91 portions of edible tulip, 3.57 to 34.09 portions of nutgrass galingale rhizome, 11.36 to 40.91 portions of Chinese honeylocust fruit, 2.38 to 34.09 portions of cochinchina momordica seed, 2.38 to 45.46 portions of common burreed rhizome, 1.79 to 40.91 portions of orange kernel and 1.79 to 38.82 portions of impatiens balsamina. The traditional Chinese medicine external preparation can improve hyperplasia of mammary glands, but does not disclose that the traditional Chinese medicine external preparation can obviously shorten the diameter of breasts of individuals with hyperplasia of mammary glands and obviously reduce the heights of breast papillae, and also does not disclose that the traditional Chinese medicine external preparation can improve endocrine disturbance and antioxidation stress, and can not meet the current requirements of scientific, accurate and stable modern traditional Chinese medicine preparations.
Therefore, searching for a more scientific and stable traditional Chinese medicine external preparation for treating hyperplasia of mammary glands by modernization is the main research direction at present.
Disclosure of Invention
In order to solve the technical problems in the prior art, the application provides a traditional Chinese medicine composition for treating hyperplasia of mammary glands, and a preparation method and application thereof. The method is realized by the following technical scheme:
a Chinese medicinal composition for treating hyperplasia of mammary glands comprises the following raw materials: radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, poria, and Natrii sulfas.
Preferably, the radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria are ethanol extracts of radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria. The method is characterized by comprising the following steps of: pulverizing the above materials into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr, stirring, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing the soaking solutions, and filtering to obtain filtrate.
Preferably, the weight ratio of the raw materials is as follows: 15-40 parts of radix ranunculi ternati, 10-35 parts of ligusticum wallichii, 10-30 parts of radix bupleuri, 5-25 parts of blumea balsamifera, 5-30 parts of green tangerine peel, 10-30 parts of poria cocos and 5-30 parts of mirabilite.
The traditional Chinese medicine composition for treating hyperplasia of mammary glands is prepared into an external preparation by adding a solvent and auxiliary materials, wherein the solvent is 30-80% ethanol, and the auxiliary materials are glycerol.
The traditional Chinese medicine composition for treating hyperplasia of mammary glands can also be prepared into other external preparations such as ointment, gel, foam, cream, spray, spirit, lotion, patch, oil agent and the like.
The preparation method of the traditional Chinese medicine composition for treating hyperplasia of mammary glands comprises the following steps:
(1) Taking radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, poria and Natrii sulfas according to a certain proportion for use;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr, stirring during soaking, collecting supernatant, soaking in 4-3 times of 30-80% ethanol for 24-72 hr, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol into the filtrate, stirring, dissolving, adding 30-80% ethanol, adjusting the amount, mixing, standing, and filtering.
Preferably, the glycerol accounts for 1-5% of the whole liquid medicine.
Preferably, each milliliter of the finally obtained liquid medicine corresponds to 0.06g-0.2g of the raw medicinal materials.
The product is topical liniment, and can be applied to skin surface for percutaneous absorption. The wettability of the skin is helpful for the absorption of the medicine, so that a proper amount of humectant is added. The glycerin is the most common humectant, has the function of absorbing water, can keep skin moist and soft, forms a glycerin film isolating air on the skin, ensures that the moisture of the skin is not easy to evaporate, can absorb the moisture in the air to moisten the skin, effectively prevents skin chapping, ensures that the covered skin cuticle is kept moist all the time, has good glycerin safety, does not contain any irritant component, and can be used with ease even if the skin is sensitive. Experiments prove that the glycerol with the concentration of 1-5% is suitable for being used as the humectant.
The traditional Chinese medicine composition for treating hyperplasia of mammary glands is applied to the aspect of being used for preparing medicines for treating hyperplasia of mammary glands.
The formula comprises the following medicinal materials:
radix Ranunculi Ternati: sweet and pungent taste and warm nature; enter liver and lung meridians; has effects of eliminating swelling, removing toxic substances, and dispelling blood stasis. The product is a dry root tuber of Maoke plant Mao Ranunculus natural Thunb. Collected in spring, removed fibrous root and silt, dried in the sun. Before feeding, selecting and removing impurities, and pulverizing into coarse powder for later use.
Bupleurum root: pungent and bitter taste, slightly cold nature, and liver, gallbladder and lung meridian entered, has the effects of dispelling heat, soothing liver, relieving depression, and lifting yang qi, and is commonly used for treating chest and hypochondrium distending pain, cold and heat, common cold and fever, etc. The product is dry root of Bupleurum scorzonerifolium Willd. Or Bupleurum scorzonerifolium of Umbelliferae. According to different characters, it is known as "Bei Hu" and nan Hu Shen "respectively. Collected in spring and autumn, removed stems, leaves and sediment, and dried. Before feeding, selecting and removing impurities, and pulverizing into coarse powder for later use.
Blumea balsamifera (blumea balsamifera) leaves: pungent taste; slightly bitter; warm nature. Has effects in dispelling pathogenic wind and removing dampness; warming middle energizer to check diarrhea; promoting blood circulation and removing toxic substances. Is commonly used for common cold due to wind-cold; headache due to head wind; rheumatalgia; diarrhea due to cold dampness; white worm disease; venomous snake bite; traumatic injury pain; tinea sore. The product is fresh or dry aerial parts of blumea balsamifera DC. And blumea balsamifera DC. Of Compositae (Bl.). Collected in summer and autumn, and dried in fresh or in shade. The product is also used for national medicine in China, impurities are removed by sorting before feeding, and the product is crushed into coarse powder for later use.
Green tangerine peel: bitter and pungent taste and warm nature; enter liver, gallbladder and stomach meridians. Has effects in dispersing stagnated liver qi, removing qi stagnation, and eliminating goiter. The product is the pericarp of dried young or immature fruit of Citrus reticulata Blanco and its cultivar of Rutaceae. Collecting self-falling young fruits 5-6 months, and sun-drying, namely 'green tangerine peel'; harvesting immature fruits 7-8 months, longitudinally cutting the fruits into four petals to a base part on the fruit peel, removing the petals, and sun-drying, so-called 'four anthocyanin peels'. Before feeding, selecting and removing impurities, and pulverizing into coarse powder for later use.
Ligusticum wallichii: pungent and warm in nature, has the effects of activating blood, relieving pain, promoting qi circulation, resolving stagnation, dispelling wind and eliminating dampness. The product is dried rhizome of Ligusticum chuanxiong Hort. Before feeding, selecting and removing impurities, and pulverizing into coarse powder for later use.
Poria cocos: sweet, bland and flat in taste, and good in effect of inducing diuresis, removing dampness, strengthening spleen and calming heart. The product is dry sclerotium of Polyporaceae fungus Poria cocos Poiaccos (Schw.) Wolf. Digging for more than 7-9 months, removing sediment after digging out, and piling up for "sweating". Spreading and airing until the surface is dry, sweating, repeating for several times until wrinkles appear and most of internal moisture is lost, and drying in the shade, namely 'Fuling Zhi'; or cutting fresh Poria into pieces according to different parts, and drying in the shade, which are called "Poria block" and "Poria tablet", respectively. Before feeding, selecting and removing impurities, and pulverizing into coarse powder for later use.
Mirabilite: salty and bitter in nature, enters stomach and large intestine meridians, and has the actions of softening hard mass, moistening dryness, purging down and relieving accumulation, and clearing heat and detumescence. The product is a crystal prepared by processing and refining sulfate mineral mirabilite. Mainly contains sodium sulfate (Na) 2 SO 4 ·10H 2 O) for standby.
The traditional Chinese medicine considers that hyperplasia of mammary glands is caused by liver depression and qi stagnation, liver and kidney deficiency and phlegm coagulation and blood stasis, and most of hyperplasia of mammary glands is caused by liver depression and blood stasis and phlegm coagulation and blood stasis. Therefore, it is indicated for liver-soothing and qi-flowing, blood-activating and stasis-resolving, phlegm-resolving, hardness-softening and stagnation-resolving. The recipe is aimed at treating liver depression and qi stagnation and phlegm coagulation and blood stasis, and the recipe takes common use of radix Ranunculi Ternati as a monarch drug, has the characteristics of pungent and warm nature, enters liver and lung channels, and has the effects of detoxification, detumescence and phlegm reduction and stagnation elimination. The bupleurum root has bitter and cold nature, enters liver, gall bladder and lung channels, has the effects of relieving exterior syndrome and fever, soothing liver and relieving depression, lifting yang qi and the like, and mainly takes the effects of soothing liver and relieving depression of the bupleurum root; blumea balsamifera is pungent and warm in nature, good in middle-jiao warming, blood circulation promoting, detoxifying, pain relieving and swelling reducing, is a common medicinal material for minority folks, and has good effects of warming middle-jiao, promoting blood circulation, relieving pain and swelling; is used as ministerial drug together with rhizoma Ligustici Chuanxiong for replenishing blood, promoting blood circulation for removing blood stasis, activating qi-flowing, relieving pain, and promoting fragrance. Taking the effects of strengthening spleen, promoting diuresis, removing dampness and resolving phlegm of poria cocos; the green tangerine peel is used as an adjuvant drug for soothing liver and regulating qi, resolving masses, removing stasis and eliminating phlegm. Natrii sulfas is effective in clearing heat, relieving swelling, softening hardness, and resolving hard mass. The medicines are compatible, and the functions of soothing liver, regulating qi, activating blood, dissolving stasis, softening hardness and resolving hard mass are collected together.
Compared with the prior art, the application has the technical effects that:
(1) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously raise the quality of individuals suffering from hyperplasia of mammary glands; the diameter of the breast is obviously shortened, and the nipple height of the breast is obviously reduced; the pathological condition of the mammary gland tissue is obviously improved.
(2) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously raise SOD activity and MDA content in mammary tissue and serum of individuals suffering from hyperplasia of mammary glands, and can restore to normal level.
(3) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously reduce the levels of E2, LH and PRL in the mammary tissue and serum of individuals suffering from hyperplasia of mammary glands, and obviously increase the level of PROG. Fully improves endocrine hormone disorder caused by hyperplasia of mammary glands and has protective effect on hyperplasia of mammary glands.
(4) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously increase protein expression of breast tissues Nrf2 and HO-1 of individuals suffering from hyperplasia of mammary glands and obviously reduce protein expression of Keap 1. The function of preventing and treating hyperplasia of mammary glands by activating the antioxidation mediated by Keap1-Nrf2/HO-1 signal path.
(5) According to the physicochemical properties of chemical components in the prescription medicine, a reasonable extraction process route is selected on the basis of fully considering the medicine characteristics of the original dosage form, and finally, the extraction is determined by adopting an immersion method, so that the modern tincture dosage form is prepared. Not only maintains the folk medication characteristics, but also overcomes a plurality of defects of traditional medication, and is developed into a health care product. The product takes Chinese herbal medicines in the province of China as raw materials, has wide crude drug sources, is favorable for the development and utilization of Chinese herbal medicine resources in the province of China, has good social and economic benefits for promoting and promoting the utilization of natural medicine resources in the province of China and developing the progress of products with high technical content and high added value, and has good social and economic benefits for the deep development of Chinese herbal medicines in the province of China.
Drawings
Fig. 1: the traditional Chinese medicine composition for treating hyperplasia of mammary glands has an influence on the weight of rats with hyperplasia of mammary glands (g, n= 8,x + -s).
Fig. 2: the application relates to an influence of a traditional Chinese medicine composition for treating hyperplasia of mammary glands on pathological forms of mammary glands (x 200, n=5), (note: K: blank control group, C: model group, R: nodules of breast eliminating tablets group, L: low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands, M: dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands, and H: high-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands.
Fig. 3: the traditional Chinese medicine composition for treating hyperplasia of mammary glands has influence (x+/-s, n=8) on SOD and MDA levels in serum and mammary tissue of a mammary gland hyperplasia model rat. P < 0.05 compared to K groups; * P < 0.01; compared to group C, #P < 0.05; # P < 0.01. And (3) injection: k: blank control group; c: a model group; r: lump eliminating tablets for breast; l: the application relates to a low-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the application relates to a dosage group in a traditional Chinese medicine composition for treating hyperplasia of mammary glands; h: the application relates to a high-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands.
Fig. 4: the traditional Chinese medicine composition for treating hyperplasia of mammary glands has influence on hormone level (x+/-s, n=8) in serum and mammary tissue of a mammary gland hyperplasia model rat. P < 0.05 compared to K groups; * P < 0.01; compared to group C, #P < 0.05; # P < 0.01. And (3) injection: k: blank control group; c: a model group; r: lump eliminating tablets for breast; l: the application relates to a low-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the application relates to a dosage group in a traditional Chinese medicine composition for treating hyperplasia of mammary glands; h: the application relates to a high-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands.
Fig. 5: the effect of the traditional Chinese medicine composition for treating hyperplasia of mammary glands on hormone level in mammary tissue of mammary gland hyperplasia model rats (x+/-s, n=8). P < 0.05 compared to K groups; * P < 0.01; compared to group C, #P < 0.05; # P < 0.01. And (3) injection: k: blank control group; c: a model group; r: lump eliminating tablets for breast; l: the application relates to a low-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the application relates to a dosage group in a traditional Chinese medicine composition for treating hyperplasia of mammary glands; h: the application relates to a high-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands.
Fig. 6: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application is used for expressing Nrf2, keap1 and HO-1 (x+/-s, n=8) in mammary tissue of a mammary gland hyperplasia model rat. P < 0.05 compared to K groups; * P < 0.01; compared to group C, #P < 0.05; # P < 0.01. And (3) injection: k: blank control group; c: a model group; r: lump eliminating tablets for breast; l: the application relates to a low-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the application relates to a dosage group in a traditional Chinese medicine composition for treating hyperplasia of mammary glands; h: the application relates to a high-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands.
Detailed Description
The technical scheme of the present application is further defined below in conjunction with the specific embodiments, but the scope of the claimed application is not limited to the description.
Example 1
(1) Taking 25g of radix ranunculi ternati, 20g of ligusticum wallichii, 15g of radix bupleuri, 15g of blumea balsamifera, 20g of green tangerine peel, 25g of poria cocos and 20g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 60% ethanol 6 times for 72 hr according to soaking method under 0120 in four general terms of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 60% ethanol 3 times for 48 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 30ml of glycerol into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 2
(1) 40g of radix ranunculi ternati, 20g of ligusticum wallichii, 15g of radix bupleuri, 10g of blumea balsamifera, 15g of green tangerine peel, 15g of poria cocos and 10g of mirabilite are taken for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 70% ethanol 6 times for 72 hr according to soaking method under 0120 of four general principles of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 70% ethanol 3 times for 48 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 30ml of glycerol into the filtrate, stirring, dissolving, adding 70% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 3
(1) Taking 30g of radix ranunculi ternati, 20g of ligusticum wallichii, 30g of radix bupleuri, 5g of blumea balsamifera, 5g of green tangerine peel, 10g of poria cocos and 5g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 80% ethanol 6 times for 72 hr according to soaking method under 0120 of four general principles of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 80% ethanol 3 times for 24 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 10ml into the filtrate, stirring, dissolving, adding 80% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 4
(1) Taking 15g of radix ranunculi ternati, 35g of ligusticum wallichii, 25g of radix bupleuri, 20g of blumea balsamifera, 30g of green tangerine peel, 20g of poria cocos and 15g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 70% ethanol 6 times for 72 hr according to soaking method under 0120 of four general principles of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 70% ethanol 3 times for 48 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 15ml into the filtrate, stirring, dissolving, adding 70% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 5
(1) Taking 30g of radix ranunculi ternati, 20g of ligusticum wallichii, 15g of radix bupleuri, 15g of blumea balsamifera, 20g of green tangerine peel, 25g of poria cocos and 30g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 5 times 30% ethanol for 120 hr according to soaking method under 0120 in four general principles of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 4 times 30% ethanol for 72 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 20ml into the filtrate, stirring, dissolving, adding 30% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 6
(1) Taking 25g of radix ranunculi ternati, 20g of ligusticum wallichii, 15g of radix bupleuri, 25g of blumea balsamifera, 15g of green tangerine peel, 30g of poria cocos and 10g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 5 times of 60% ethanol for 96 hr according to soaking method under general rule 0120 of four parts of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 4 times of 60% ethanol for 72 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 50ml into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
Example 7
(1) Taking 25g of radix ranunculi ternati, 10g of ligusticum wallichii, 30g of radix bupleuri, 15g of blumea balsamifera, 15g of green tangerine peel, 20g of poria cocos and 20g of mirabilite for standby;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, and Poria into coarse powder, soaking in 60% ethanol 6 times for 72 hr according to soaking method under 0120 in four general terms of Chinese pharmacopoeia 2020 edition, stirring during soaking, collecting supernatant, soaking in 60% ethanol 3 times for 72 hr, collecting supernatant, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 40ml of glycerol into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting the dosage to 1000ml, mixing, standing, and filtering.
The products of the embodiments of the application were tested by the toxicity test center of the university of Guizhou medical science. Physical and chemical index test results: mercury (Hg) is <1mg/L, arsenic (As) is <2mg/L, lead (pb) is <10mg/L, cadmium (Cd) is <5mg/L, and methanol is <2000mg/L, which meet the standard specification of the national food and drug administration (cosmetic safety technical Specification (2015 edition); microbial index test results: colony count (CFU/mL) <10, mold and yeast (CFU/mL) <10, heat-resistant Escherichia coli, pseudomonas aeruginosa and Staphylococcus aureus were not detected, and the method meets the standard specification of the national food and drug administration (cosmetic safety technical Specification (2015 edition); toxicology test results: the acute oral toxicity test result shows that LD50 is more than 5000mg/kg, and the oral toxicity test result is practically nontoxic; the results of multiple skin irritation tests show that the integral average value is 0, the skin irritation index is 0-0.5, and the skin irritation intensity is non-irritating. Meets the standard specification of the national food and drug administration (2015 edition); the above test results show that the product meets the national relevant standards and is safe to use.
The applicant performs the study on the effect and mechanism of the anti-hyperplasia of mammary glands on the traditional Chinese medicine composition obtained by each embodiment of the application, and experimental results prove that the traditional Chinese medicine composition obtained by each embodiment has the effect of preventing and treating the hyperplasia of mammary glands on rats with mammary glands models, can regulate the levels of hormones E2, LH, PRL and PROG, raise SOD activity, reduce MDA content and achieve the effect of preventing and treating hyperplasia of mammary glands by activating Keap1-Nrf2/HO-1 signal paths.
The applicant discloses by taking the specific scheme of the anti-hyperplasia effect and mechanism research of the embodiment 1 as an example, and the specific scheme is as follows:
study of the effects and mechanisms of the Chinese medicinal composition for treating hyperplasia of mammary glands of example 1
1 materials and methods
1.1 test drug
The application relates to a traditional Chinese medicine composition for treating hyperplasia of mammary glands, which comprises the following components: the external preparation of the traditional Chinese medicine composition prepared by the method of the example 1; nodules of breast tablet (Lot: 200110): liaoning nurses (groups) Limited; estradiol benzoate (Lot #: C11924650), progesterone (Lot #: C12082401): shanghai Miclin Biochemical technologies Co., ltd; 4% paraformaldehyde (Lot: 0708A 22): beijing Lei Gen Biotechnology Co., ltd; superoxide dismutase SOD (Lot: 20220214), malondialdehyde MDA (Lot: 20220414), rat estradiol E2 (Lot: 20220217), rat progesterone PROG (Lot: 20211219), rat luteinizing hormone LH (Lot: 20220515), rat prolactin PRL (Lot: 20220518), lysate (RIPA), BCA protein concentration determination kit, marker, beta-actin polyclonal antibody, goat anti-rabbit IgG, goat anti-mouse IgG, tris, SDS and glycine: soy Biotechnology Co., ltd; PVDF film: millipore Trading Co.ltd; mouse monoclonal antibody Keap1 (ab 119403), rabbit polyclonal antibody Nrf2 (ab 156883), rabbit polyclonal antibody HO-1 (ab 137749).
1.2 instruments
Vernier caliper: shanghai Hui Yi Ji Zhi Ling Co., ltd; leictp 1020 automatically prevents the dehydrator: leica embedding machine: leica cm2245 rotary paraffin microtome: optical microscope: the Leica company, germany; an electronic balance: sidoriscom instruments (Beijing); -80 ℃ low temperature refrigerator: sanyang Japan; milli QA pure water processor: millipo tracking Co.ltd, france; W-80A type vortex mixer: medical instrument factory of kylin of Jiangsu sea gate; enzyme-labeled instrument: BIO-RAD Co., USA; and (3) an oven: shanghai precision macro laboratory equipment Co., ltd; centrifuge: eppendorf, germany.
1.3 laboratory animals
Female Sprague Dawley (SD) rats purchased from Beijing Fukang Biotech Co., ltd (license number SCXK (Beijing) 2019-0008).
1.4 grouping and administration
Female SD rats were randomized into 6 groups of 10: blank control group (K), model control group (C), nodules of breast eliminating tablet group (R), the traditional Chinese medicine composition for treating hyperplasia of mammary glands of the application is low (L), medium (M) and high dose group (H). Except for the blank control group, the rats of each group replicate the mammary gland hyperplasia model of the rats according to the literature, and the inner side of the hind leg is intramuscularly injected with estradiol benzoate (0.5 mg/kg) 1 time per day for 25 days, followed by intramuscular injection of progesterone (5 mg/kg) 1 time per day for 5 days. At the same time of molding, normal saline (1 mL/100 g) is administrated by gastric lavage of the blank control group and the model control group, the administration dosage of the breast nodule eliminating tablet group is 960mg/kg, and the low, medium and high dosage groups of the traditional Chinese medicine composition for treating hyperplasia of mammary glands are respectively smeared with 1, 2 and 3 times of medicines every day for 45 days.
1.5 detection index
After 24 hours of last dose, (1) the 2 nd pair breast diameter and nipple height of the rat were precisely measured with a vernier caliper. (2) Hematoxylin-eosin (HE) staining: after the rat was syncoped, the rat was quickly fixed on a rat plate in supine position after anesthesia with 7% chloral hydrate by intraperitoneal injection (0.5 mL/100 g), then the mammary gland of rat # 2 was fixed with 4% paraformaldehyde for 2d, tissue dehydration, paraffin embedding, and finally sections were HE stained. And taking 3 rd and 4 th to perform biochemical indexes, ELISA and WB detection on the mammary gland of the breast.
2 experimental results
2.1 effects of the Chinese medicinal composition for treating hyperplasia of mammary glands on weight change of rats
The rats had a higher body weight, and the rats in the blank group had a higher body weight, and the rats in the other model groups had a lower body weight, and after 21 days of administration, the weight gain of the nodules of breast tablet group and each dose of the drug group was more remarkable than that of the model group (fig. 1).
2.2 effects of the Chinese medicinal composition for treating hyperplasia of mammary glands of the present application on nipple height and breast diameter
Compared with the blank control group, the nipple height and the breast diameter of the model control group are obviously increased, and the model control group has statistical significance (P < 0.05). Compared with a model control group, the mammary nodule eliminating tablet group, the traditional Chinese medicine composition for treating hyperplasia of mammary glands, and the high-dose group can obviously reduce the nipple height and the mammary gland diameter of rats with hyperplasia of mammary glands caused by estradiol benzoate and progesterone, and have statistical significance (P is less than 0.05, see table 1).
TABLE 1 Effect of the Chinese medicinal composition for treating mammary gland hyperplasia of the present application on mammary gland height and diameter of mammary gland hyperplasia rats
In contrast to the K groups, * P﹤0.05； ** p < 0.01; in contrast to the group C, # P﹤0.05； ## p < 0.01. And (3) injection: k: blank control group; c: a model group; r: lump eliminating tablets for breast; l: the application relates to a low-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the application relates to a dosage group in a traditional Chinese medicine composition for treating hyperplasia of mammary glands; h: the application relates to a high-dose group of traditional Chinese medicine composition for treating hyperplasia of mammary glands.
2.3 effects of the Chinese medicinal composition for treating hyperplasia of mammary glands on pathological morphology of mammary tissue
Acinus and lobules of mammary tissue of rats in the blank group were normal; the number of lobules of mammary glands of rats in the model group is obviously increased, a large amount of secretions appear when the glandular cavities of the mammary glands are expanded, and epithelial cells of the catheter are arranged in disorder and obviously proliferate; compared with the model group, the positive medicine group, the traditional Chinese medicine composition for treating hyperplasia of mammary glands, provided by the application, have the advantages that the hyperplasia of mammary tissue of each dosage administration group and the stomach irrigation administration group is improved, the number of acinus is obviously reduced, the volume of gland lobules is obviously reduced, and the secretion of ducts and cavities of the mammary glands is reduced.
The experimental result shows that compared with a model group, the quality of the rat body of each dose of the traditional Chinese medicine composition for treating hyperplasia of mammary glands is obviously improved; the diameter of the breast of the rat is obviously shortened, and the height of the nipple of the breast is obviously reduced; the pathological condition of mammary tissue is obviously improved.
2.4 influence of the Chinese medicinal composition for treating hyperplasia of mammary glands on MDA content and SOD activity in mammary tissue and serum of rats with hyperplasia of mammary glands
The hyperplasia of mammary glands induced by the estradiol benzoate and the progesterone can cause oxidative damage, and the SOD activity and MDA content in serum and mammary tissues are detected to evaluate the protective effect of the traditional Chinese medicine composition for treating hyperplasia of mammary glands on the oxidative damage of hyperplasia of mammary glands. As shown in fig. 3, the SOD activity in mammary tissue and serum of rats in model group (C) was significantly reduced, while the MDA content was significantly increased (P < 0.01) compared to the blank group (K); after administration, the Chinese medicinal composition for treating hyperplasia of mammary glands has obvious improvement effect on SOD activity and MDA content in rat mammary tissue and serum, and the high dose group of the Chinese medicinal composition for treating hyperplasia of mammary glands has the effect of recovering the SOD activity and MDA content in rat mammary tissue and serum to the level of a blank control group.
2.5 effects of the Chinese medicinal composition for treating hyperplasia of mammary glands of the application on hormone levels in mammary tissue and serum of rats with hyperplasia of mammary glands
As shown in fig. 4 and 5, the levels of E2, LH and PRL in mammary tissue and serum were significantly increased (P < 0.05) and the levels of PROG were significantly decreased (P < 0.05) in the rats in model group (C) compared to the blank group (K); compared with the model group, after the Chinese medicinal composition for treating hyperplasia of mammary glands is administered, the levels of E2, LH and PRL in mammary tissue and serum of rats are obviously reduced (P is less than 0.05), and the levels of PROG are obviously increased (P is less than 0.05).
2.6 the Chinese medicinal composition for treating hyperplasia of mammary glands of the application expresses Nrf2, keap1 and HO-1 proteins in mammary tissue of rats with hyperplasia of mammary glands
As shown in FIG. 6, the results of immunoblotting method for detecting protein expression of Nrf2, keap1 and HO-1 in the mammary tissue of each group of rats show that compared with the blank control group (K), the protein expression of Nrf2 and HO-1 in the mammary tissue of the rat in the model group (C) is obviously reduced (P < 0.01), and the protein expression of Keap1 is obviously increased (P < 0.01); compared with the model group, after administration of the Chinese medicinal composition for treating hyperplasia of mammary glands, protein expression of rat mammary tissue Nrf2 and HO-1 is obviously increased (P is less than 0.01), and protein expression of Keap1 is obviously reduced (P is less than 0.01).
Discussion 3
Hyperplasia of mammary glands is caused by endocrine disturbance, and has close relation with in vivo hormone disturbance, especially estrogen and progestogen proportion unbalance. PRL is a prolactin produced by eosinophils in the distal part of the pituitary, and studies have shown that: PRL has effects of promoting mammary gland growth and development and maintaining lactation, and can inhibit hypothalamus-pituitary-ovary axis. When PRL in mammary tissue and blood is abnormally increased, the PRL can inhibit the secretion of PROG, and simultaneously stimulate the synthesis of E2, so that the E2/PROG ratio is disordered, and the PRL can produce bad stimulation on the mammary tissue to cause the hyperplasia of the mammary tissue. E2 can promote regeneration of the mammary duct, and its expression can inhibit the anterior pituitary gland from secreting LH, so that on one hand, the growth of the mammary duct is inhibited, and on the other hand, the development of mammary lobules is inhibited to a certain extent. The results of this study showed that: the traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously reduce the levels of mammary tissue and serum E2, LH and PRL of a mammary gland hyperplasia model rat, and increase the level of PROG. The endocrine hormone disturbance of the mammary gland hyperplasia model rat is improved, and the result of the mammary gland tissue pathological section is combined, so that the traditional Chinese medicine composition for treating the mammary gland hyperplasia has a protective effect on the mammary gland hyperplasia.
In addition, oxidative stress is also an important factor of hyperplasia of mammary glands, after estradiol benzoate and progesterone induce hyperplasia of mammary glands, SOD activity in rat serum and mammary tissue is obviously reduced, and MDA content is obviously increased. The imbalance of endogenous oxidative status results in activation of the antioxidant defense system, and the Keap1-Nrf2/HO-1 signaling pathway is a newly discovered defensive transduction pathway of the body against external oxidative stimuli in recent years. Normally, the combination of Nrf2 and a cytoskeletal associated protein Keap1 exists in cytoplasm in the form of dimer, after being stimulated by external oxidative stress factors, the Nrf2 and the Keap1 are dissociated and transformed into nuclei, and then the proteins such as antioxidant protein HO-1 are induced to express by interaction with an antioxidant stress response element AR, so that the antioxidant effect is exerted. The Westernblot detects the protein contents of Nrf2, keap1 and HO-1 in mammary tissue, and the result shows that compared with a blank control group, the protein expression amounts of Nrf2 and HO-1 in mammary tissue of a model group are obviously reduced, and the protein expression amount of Keap1 is increased (P < 0.01); compared with the model group, each dosing group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously increase the protein expression quantity of Nrf2 and HO-1 and reduce the protein expression quantity of Keap1 (P < 0.01). The traditional Chinese medicine composition for treating hyperplasia of mammary glands is possibly used for preventing and treating hyperplasia of mammary glands by activating the antioxidation mediated by Keap1-Nrf2/HO-1 signal path.
Conclusion 4
The traditional Chinese medicine composition for treating hyperplasia of mammary glands has good curative effect on rats with models of hyperplasia of mammary glands, and possible mechanisms of the traditional Chinese medicine composition can be used for adjusting the levels of hormones E2, LH, PRL and PROG, increasing SOD activity, reducing MDA content and achieving the effect of preventing hyperplasia of mammary glands by activating Keap1-Nrf2/HO-1 signal paths.
Finally, it should be noted that the above embodiments are merely representative examples of the present application. Obviously, the technical solution of the application is not limited to the above-described embodiments, but many variations are possible. All modifications directly derived or suggested to one skilled in the art from the present disclosure should be considered as being within the scope of the present application.
Claims (9)
1. The traditional Chinese medicine composition for treating hyperplasia of mammary glands is characterized by being prepared from the following raw materials in parts by weight: 15-40 parts of radix ranunculi ternati, 10-35 parts of ligusticum wallichii, 10-30 parts of radix bupleuri, 5-25 parts of blumea balsamifera, 5-30 parts of green tangerine peel, 10-30 parts of poria cocos and 5-30 parts of mirabilite.
2. The Chinese medicinal composition for treating hyperplasia of mammary glands according to claim 1, wherein the radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos are ethanol extracts of radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos.
3. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 2, wherein the ethanol extracts of radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos are extracted by the following method: pulverizing the above materials into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr, stirring, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing the soaking solutions, and filtering to obtain filtrate.
4. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 1, wherein a solvent and auxiliary materials are also added to prepare an external preparation.
5. The Chinese medicinal composition for treating hyperplasia of mammary glands according to claim 4, wherein the solvent is 30-80% ethanol and the adjuvant is glycerol.
6. The method for preparing the traditional Chinese medicine composition for treating hyperplasia of mammary glands according to any one of claims 1 to 5, comprising the following steps:
(1) Taking radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, poria and Natrii sulfas according to a certain proportion for use;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr, stirring during soaking, collecting supernatant, soaking in 4-3 times of 30-80% ethanol for 24-72 hr, mixing the soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol into the filtrate, stirring, dissolving, adding 30-80% ethanol, adjusting the amount, mixing, standing, and filtering.
7. The method for preparing a traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 6, wherein the glycerol accounts for 1-5% of the whole liquid medicine.
8. The method for preparing a traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 6, wherein the final liquid medicine is 0.06g-0.2g of the corresponding raw medicinal materials per milliliter of the liquid medicine.
9. The use of a Chinese medicinal composition for treating hyperplasia of mammary glands according to any one of claims 1 to 5 for preparing a medicament for treating hyperplasia of mammary glands.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104436046A (en) * | 2014-11-30 | 2015-03-25 | 顾蔷怡 | Medicine for treating hyperplasia of mammary glands and preparation method thereof |
| CN107362257A (en) * | 2016-05-11 | 2017-11-21 | 黄名玉 | A kind of medicine for treating the proliferation of mammary gland |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104436046A (en) * | 2014-11-30 | 2015-03-25 | 顾蔷怡 | Medicine for treating hyperplasia of mammary glands and preparation method thereof |
| CN107362257A (en) * | 2016-05-11 | 2017-11-21 | 黄名玉 | A kind of medicine for treating the proliferation of mammary gland |
Non-Patent Citations (2)
| Title |
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| 中西医结合辨证治疗乳腺增生60例及护理;秋荣;姚明松;;中国民间疗法;-;第-卷(第08期);第69-70页 * |
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