CN115919939A - 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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- CN115919939A CN115919939A CN202211548831.3A CN202211548831A CN115919939A CN 115919939 A CN115919939 A CN 115919939A CN 202211548831 A CN202211548831 A CN 202211548831A CN 115919939 A CN115919939 A CN 115919939A
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Abstract
The invention 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 radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride, poria cocos and mirabilite as raw materials, and takes the radix ranunculi ternati as a monarch drug. The main action of chai Hu is to soothe the liver and relieve depression; the blumea balsamifera has pungent and warm nature and flavor, is good at warming middle-jiao and activating blood, and detoxifying, relieving pain and diminishing swelling, and has better effects of warming middle-jiao and activating blood, and relieving pain and diminishing swelling; and rhizoma Ligustici Chuanxiong has effects of replenishing blood, promoting blood circulation for removing blood stasis, activating qi-flowing, relieving pain, and removing aromatic odor. Then taking the tuckahoe to invigorate the spleen, promote diuresis, eliminate dampness, reduce phlegm and the like; it is used as adjuvant drug together with Qing Pi for soothing liver, regulating qi, dissipating stagnation, removing blood stasis and eliminating phlegm. Mirabilite is good at clearing fire, relieving swelling, softening hardness and dissipating nodulation. The medicines are compatible to play the roles of soothing the liver, regulating qi, promoting blood circulation, removing blood stasis, softening hardness and dissipating stagnation. Finally, the modern traditional Chinese medicine external preparation with good treatment effect on the hyperplasia of mammary glands is obtained.
Description
Technical Field
The invention 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-tumor disease of women, and related investigation shows that the hyperplasia of mammary glands is the most common disease of mammary glands and has the characteristics of long course of disease, slow development, easy relapse and the like. According to statistics, the incidence rate of women of childbearing age is more than 60%. With the acceleration of social life rhythm and the reasons of environmental pollution and the like in recent years, the prevalence rate shows a trend of increasing year by year in recent years, the clinical symptoms mainly comprise breast distending pain, breast lumps and nipple discharge, economic and mental pressure is brought to female patients, the physical and mental health of the patients is seriously affected, and the social importance is more and more attracted.
The western medicine considers that the incidence of hyperplasia of mammary glands is related to endocrine dyscrasia and mental factors, particularly to balance disorder of hypothalamus-pituitary-ovary-mammary gland endocrine axes, the dysfunction of ovary secretion causes hypoxanthine secretion, relative increase of estrogen, and finally causes hyperplasia of mammary gland epithelium and fibrous tissue, degenerative change of mammary gland tissue ducts and mammary lobules in structure and progressive connective tissue growth. Most western medicines adopt hormones, vitamins, iodine agents and other symptomatic treatment medicines, and the medicines can regulate the level of female and pregnant hormones in vivo and relieve pain, but the medicines have great disputes because of the phenomena of hot flashes, night sweats, headaches, irritability, low menstrual flow and even menopause, unstable curative effect and the like after the medicines are taken by a high proportion of patients, so that the western medicines are not preferred.
According to traditional Chinese medicine, mammary gland hyperplasia belongs to the range of mammary nodules in traditional Chinese medicine, and the pathogenesis of the mammary gland hyperplasia is as follows: the term "ren Chong and ren Chong are the root causes of pain and core formation due to qi stagnation, accumulation in breast space, or hard or swollen, if the regulation is lost, or wind pathogen attacks. The patients cause depression of liver-qi, deficiency of liver and kidney, disorder of thoroughfare and conception vessel, phlegm coagulation and blood stasis due to emotional, fatigue, diet and physique, and finally cause hyperplasia of mammary glands. Hyperplasia of mammary glands is often manifested as breast pain and breast palpation with nodules that become more pronounced and aggravated before menstruation. The hyperplasia of mammary glands has various pathological types, if the regulation and treatment are not actively carried out, the physical and mental pains and pressure are brought to patients, and the cancer can be seriously worsened. According to the traditional Chinese medicine, the breast diseases are closely related to the channels of the liver, the stomach, the Chong channel, the ren channel and the like. Mammary gland caking diseases are caused by unsmooth daily life or work, high pressure, excessive anxiety and anger, liver and spleen damage, liver depression and qi stagnation and spleen and stomach disorder, the spleen and stomach can not normally transport and transform water dampness, so that the water dampness is accumulated into phlegm, and if the phlegm and the qi are mutually bound in breasts and the stasis and the stagnation are blocked, nodules of breast are formed. Some patients are also caused by yang deficiency and phlegm-dampness accumulation due to the disorder of chong and ren meridians caused by the deficiency of liver and kidney.
The traditional Chinese medicine external preparation for treating hyperplasia of mammary glands at present has the problems of extensive process, unstable quality, incapability of ensuring the consistency and stability of medication, inaccurate medication dosage, large 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 to 12 parts of epimedium, 8 to 12 parts of cistanche, 3 to 7 parts of cornu cervi degelatinatum, 8 to 12 parts of radix bupleuri, 8 to 12 parts of rhizoma cyperi, 8 to 12 parts of angelica, 8 to 12 parts of white peony root, 8 to 12 parts of ligusticum wallichii, 3 to 7 parts of bulbus fritillariae cirrhosae, 8 to 12 parts of selfheal, 8 to 12 parts of fructus trichosanthis, 3 to 7 parts of rhubarb, 3 to 7 parts of mirabilite, 3 to 7 parts of myrrh, 8 to 12 parts of seaweed, 8 to 12 parts of kelp and 3 to 7 parts of borneol; processing the above Chinese medicinal materials, and making into patch. The preparation method does not comprise the steps of separation, extraction, purification and the like, has extensive process and unstable quality, and the prepared patch has poor absorption effect and needs to be used together with equipment such as a massager and the like.
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 ternate buttercup root, 1.79 to 34.09 portions of rhizoma corydalis, 5.35 to 45.46 portions of muskroot-like semiaquilegia root, 1.79 to 40.91 portions of Indian iphigenia bulb, 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 tangerine seed and 1.79 to 38.82 portions of garden balsam seed. The traditional Chinese medicine preparation can improve hyperplasia of mammary glands, but does not disclose that the diameter of the breast of a hyperplasia of mammary glands individual can be obviously shortened, the height of the breast nipple is obviously reduced, and the endocrine disturbance and the anti-oxidative stress effect can be improved.
Therefore, the search for a more scientific, stable and modern external traditional Chinese medicine preparation for treating hyperplasia of mammary glands is the main research direction at present.
Disclosure of Invention
In order to solve the technical problems in the prior art, the invention 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 traditional Chinese medicine composition for treating hyperplasia of mammary glands comprises the following raw material components: radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, poria, and Natrii sulfas.
Preferably, the catclaw buttercup root, the ligusticum wallichii, the radix bupleuri, the blumea balsamifera, the green tangerine peel and the poria cocos are ethanol extracts of the catclaw buttercup root, the ligusticum wallichii, the radix bupleuri, the blumea balsamifera, the green tangerine peel and the poria cocos. The method is specifically obtained by extracting the following steps: pulverizing the raw materials into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr while stirring, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing 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 pericarpium citri reticulatae viride, 10-30 parts of poria cocos and 5-30 parts of mirabilite.
The traditional Chinese medicine composition for treating hyperplasia of mammary glands is further added with a solvent and an auxiliary material to prepare an external preparation, wherein the solvent is 30-80% ethanol, and the auxiliary material is 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 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 at 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, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol into the filtrate, stirring, dissolving, adding 30-80% ethanol, adjusting to desired volume, 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 liniment for external use, and can be applied on skin surface for percutaneous absorption. The skin wettability is helpful for the absorption of the medicine, so a proper amount of humectant is added. The glycerin is the most common moisturizing agent, has the water absorption effect, can keep the skin moist and soft, forms a glycerin film for isolating the 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 the skin from being dried and cracked, ensures that the stratum corneum covered by the glycerin film can be kept moist all the time, has good glycerin safety, does not contain any irritant component, and can be used safely even if the skin is sensitive. It is suitable to add 1-5% glycerin as humectant.
The traditional Chinese medicine composition for treating hyperplasia of mammary glands is applied to the aspect of being used as/preparing a medicine for treating hyperplasia of mammary glands.
The formula comprises the following medicinal materials:
radix ranunculi ternati: sweet and pungent taste, warm in nature; it enters liver and lung meridians; has effects in dissipating swelling, removing toxic materials, and dispelling blood stasis. The product is dry root tuber of Maoke plant Mao rannunculus ernatus thunb. Collected in spring, removed fibrous root and silt, and dried in the sun. Before feeding, selecting to remove impurities, and pulverizing into coarse powder for use.
Bupleurum root: it is pungent and bitter in flavor, slightly cold in nature, entering liver, gallbladder and lung meridians, and has the effects of dispelling heat, relieving liver qi stagnation, and lifting yang qi, and is commonly used for treating distending pain in chest and hypochondrium, alternating cold and heat, common cold and fever, etc. The product is dried root of Bupleurum chinense DC or Bupleurum scorzonerifolium Willd. According to the different characteristics, it is called "Bei chai Hu" and "nan chai Hu" respectively. Collected in spring and autumn, removed stems and leaves and silt, and dried. Before feeding, selecting to remove impurities, and pulverizing into coarse powder for use.
Blumea balsamifera: 2, pungent taste; slightly bitter; and (4) temperature. Has the functions of dispelling wind and eliminating dampness; warming middle energizer to arrest diarrhea; promoting blood circulation and removing toxic substance. It is commonly used for common cold due to wind-cold; headache due to wind-heat; rheumatic arthralgia; dysentery due to cold-dampness; cunninghamiasis; bite by poisonous snake; traumatic injury and pain; tinea sore. The product is fresh or dried aerial parts of blumea balsamifera (L.) DC. And blumea pseudoabundana (Bl.) DC. Of Compositae. Collected in summer and autumn, and used fresh or dried in the shade. The product is also used as a medicine for minority nationality in our province, and is selected to remove impurities before feeding, and is crushed into coarse powder for later use.
Green tangerine peel: bitter and pungent taste, warm in nature; enter liver, gallbladder and stomach meridians. Has effects in dispersing depressed liver-energy, relieving qi stagnation, and eliminating goiter. The product is pericarp of dry young fruit or immature fruit of Citrus reticulata blanco of Rutaceae and its cultivar. Collecting the self-falling young fruits for 5-6 months, and drying in the sun, wherein the young fruits are called as 'green tangerine peels'; immature fruits are harvested in 7-8 months, longitudinally cut into four petals on the peel to the base, the petals are removed completely, and the peel is dried in the sun, and is called as 'Sihua Qing Pi'. Before feeding, selecting to remove impurities, and pulverizing into coarse powder for use.
Ligusticum wallichii: pungent taste and warm nature, and has effects of promoting blood circulation, relieving pain, activating qi-flowing, resolving stagnation, dispelling pathogenic wind and removing dampness. The product is dried rhizome of Ligusticum chuanxiong Hort. Before feeding, selecting to remove impurities, and pulverizing into coarse powder for use.
Tuckahoe, poria cocos: sweet, bland and neutral in taste, and has the effects of promoting diuresis, eliminating dampness, invigorating spleen and calming heart, and can be used for treating heart diseases. The product is dried sclerotium of Poriacocos (Schw.) Wolf of Polyporaceae. Digging more than 7-9 months, removing silt after digging, and piling for sweating. Spreading and drying in the air until the surface is dry, then sweating, repeating for several times until wrinkles appear and most of the internal water is lost, and drying in the shade, which is called as 'Poria cocos'; or cutting fresh Poria according to different parts, and drying in the shade to obtain Poria block and Poria tablet. Before feeding, selecting to remove impurities, and pulverizing into coarse powder for use.
Mirabilite: with cold nature, salty and bitter taste, entering stomach and large intestine meridians, it has the actions of softening hardness, moistening dryness, purging and removing accumulation, clearing heat and relieving swelling. The product is crystal of sulfate mineral mirabilite, which is refined by processing. Sodium sulfate (Na) mainly containing water 2 SO 4 ·10H 2 O), and standby.
According to the traditional Chinese medicine, mammary gland hyperplasia is caused by liver depression and qi stagnation, liver and kidney deficiency, phlegm coagulation and blood stasis, and most of the mammary gland hyperplasia is caused by liver depression followed by blood stasis, and phlegm coagulation is formed into blocks. Therefore, soothing liver and regulating qi, activating blood and dissolving stasis, resolving phlegm, softening hardness and dissipating stagnation are the treatment principles. The prescription is used for treating liver depression and qi stagnation and phlegm coagulation and blood stasis, and the common medicine of the minority is ternate buttercup root in the prescription as a monarch medicine, and the ternate buttercup root is pungent and warm in nature and enters liver and lung channels, so that the prescription has the effects of detoxifying, reducing swelling and resolving phlegm and resolving masses. The bupleurum has bitter and cold property, enters liver, gallbladder and lung channels, has the efficacies of relieving exterior syndrome, reducing fever, soothing liver-qi stagnation, lifting yang qi and the like, and the formula mainly takes the effect of soothing liver-qi stagnation; the blumea balsamifera has pungent and warm nature and good effects of warming the middle-jiao and activating blood, and detoxifying, relieving pain and diminishing swelling, is a frequently used medicinal material for minority nationalities, and has better effects of warming the middle-jiao and activating blood, relieving pain and diminishing swelling; and rhizoma Ligustici Chuanxiong has effects of replenishing blood, promoting blood circulation for removing blood stasis, activating qi-flowing, relieving pain, and removing aromatic odor. Then taking the tuckahoe to invigorate the spleen, promote diuresis, eliminate dampness, reduce phlegm and the like; pericarpium Citri Reticulatae viride has the effects of soothing liver, regulating qi, dissipating stagnation, removing blood stasis and eliminating phlegm as adjuvant drugs. Mirabilite is good at clearing fire, relieving swelling, softening hardness and dissipating nodulation. The medicines are compatible to achieve the effects of soothing liver, regulating qi, promoting blood circulation, removing blood stasis, softening hardness and dissipating stagnation.
Compared with the prior art, the invention has the technical effects that:
(1) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously improve the physique of individuals suffering from hyperplasia of mammary glands; the diameter of the breast is obviously shortened, and the height of the nipple of the breast is obviously reduced; the pathological condition of the mammary tissue is obviously improved.
(2) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously increase the SOD activity and the MDA content in mammary tissues and serum of individuals suffering from hyperplasia of mammary glands, and can recover to a 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 mammary tissues and serum of individuals suffering from hyperplasia of mammary glands and obviously increase the level of PROG. Can be used for improving endocrine hormone disorder caused by hyperplasia of mammary glands, and protecting hyperplasia of mammary glands.
(4) The traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously improve the protein expression of the Nrf2 and HO-1 of mammary tissues of individuals with hyperplasia of mammary glands and obviously reduce the protein expression of Keap 1. Preventing and treating hyperplasia of mammary glands by activating the antioxidant effect mediated by a Keap1-Nrf2/HO-1 signal channel.
(5) According to the physicochemical properties of chemical components in the medicinal taste of the prescription, on the basis of fully considering and reserving the medication characteristics of the original preparation form, a reasonable extraction process route is selected, and finally, the extraction is determined to be adopted by an immersion method to prepare the modern tincture preparation form. Not only retains the characteristic of folk medicine application, but also overcomes a plurality of defects of the traditional medicine application, and is developed into a health care product. The product takes Chinese herbal medicines of our province as raw materials, has wide raw medicine sources, is beneficial to the development and utilization of Chinese herbal medicine resources of our province, promotes the progress of the advantage utilization of natural medicine resources of our province and the development of products with high technical content and high added value, and can generate good social benefit and economic benefit for the deep development of Chinese herbal medicines of our province.
Drawings
FIG. 1: the application of the traditional Chinese medicine composition for treating hyperplasia of mammary glands has the effect on the body weight of rats with hyperplasia of mammary glands (g, n =8,x +/-s).
FIG. 2: the influence of the traditional Chinese medicine composition for treating the hyperplasia of mammary glands on the pathological form of the mammary gland tissue (× 200,n = 5) (note: K: a blank control group, C: a model group, R: a nodules of breast tablet group, L: a traditional Chinese medicine composition low-dose group for treating the hyperplasia of mammary glands, M: a traditional Chinese medicine composition medium-dose group for treating the hyperplasia of mammary glands and H: a traditional Chinese medicine composition high-dose group for treating the hyperplasia of mammary glands).
FIG. 3: the traditional Chinese medicine composition for treating hyperplasia of mammary glands has the influence on the levels of SOD and MDA in the serum and the mammary gland tissues of a mammary gland hyperplasia model rat (x +/-s, n = 8). P < 0.05 in comparison with group K; * P < 0.01; compared with group C, # P < 0.05; # P < 0.01. Note: k: blank control group; c: a model group; r: mammary lump eliminating tablet; l: the low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands of the application; m: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application comprises a dosage group; h: the application relates to a traditional Chinese medicine composition high-dose group for treating hyperplasia of mammary glands.
FIG. 4: the influence of the traditional Chinese medicine composition for treating hyperplasia of mammary glands on the hormone level in the serum and the mammary gland tissue of a mammary gland hyperplasia model rat (x +/-s, n = 8) is disclosed. P < 0.05 in comparison with group K; * P < 0.01; compared with group C, # P < 0.05; # P < 0.01. Note: k: blank control group; c: a model group; r: mammary lump eliminating tablet; l: the low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands of the application; m: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application comprises a dosage group; h: the application relates to a traditional Chinese medicine composition high-dose group for treating hyperplasia of mammary glands.
FIG. 5: the application of the traditional Chinese medicine composition for treating hyperplasia of mammary glands has the effect on the hormone level in the mammary gland tissue of a mammary gland hyperplasia model rat (x +/-s, n = 8). P < 0.05 in comparison with group K; * P < 0.01; compared with group C, # P < 0.05; # P < 0.01. Note: k: blank control group; c: a model group; r: breast nodule eliminating tablet; l: the low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands of the application; m: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application comprises a dosage group; h: the application relates to a traditional Chinese medicine composition high-dose group 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 treating the expression conditions of Nrf2, keap1 and HO-1 in mammary gland tissues of mammary glands of mammary gland hyperplasia model rats (x +/-s, n = 8). P < 0.05 in comparison with group K; * P < 0.01; # P < 0.05 compared to group C; and # P < 0.01. Note: k: blank control group; c: a model group; r: mammary lump eliminating tablet; l: the low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application comprises a dosage group; h: the application relates to a traditional Chinese medicine composition high-dose group for treating hyperplasia of mammary glands.
Detailed Description
The technical solution of the present invention is further defined below with reference to specific embodiments, but the scope of protection 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 pericarpium citri reticulatae viride, 25g of poria cocos and 20g of mirabilite for later use;
(2) Crushing radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos into coarse powder, soaking for 72 hours by 6 times of 60% ethanol according to a soaking method under 0120 of the general rule of the four parts of the 2020 edition of Chinese pharmacopoeia, stirring in the soaking process, pouring out supernate, adding 3 times of 60% ethanol for soaking for 48 hours, pouring out supernate, combining soaking liquid and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 30ml into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting to 1000ml, mixing, standing, and filtering.
Example 2
(1) Taking 40g of radix ranunculi ternati, 20g of ligusticum wallichii, 15g of radix bupleuri, 10g of blumea balsamifera, 15g of pericarpium citri reticulatae viride, 15g of poria cocos and 10g of mirabilite for later use;
(2) Crushing radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos into coarse powder, soaking for 72 hours in 70% ethanol in 6 times according to a soaking method in 0120 item of the general rule of the four parts of the 2020 edition of Chinese pharmacopoeia, stirring in the soaking process, pouring out supernate, adding 70% ethanol in 3 times for soaking for 48 hours, pouring out supernate, combining soaking liquid and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 30ml into the filtrate, stirring, dissolving, adding 70% ethanol, adjusting 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 pericarpium citri reticulatae viride, 10g of poria cocos and 5g of mirabilite for later use;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria into coarse powder, soaking with 6 times of 80% ethanol for 72 hr according to 0120 of 2020 th ministerial general rule of Chinese pharmacopeia, stirring during soaking, collecting supernatant, adding 3 times of 80% ethanol, soaking for 24 hr, collecting supernatant, mixing soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 10ml glycerol into the filtrate, stirring, dissolving, adding 80% ethanol, adjusting 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 pericarpium citri reticulatae viride, 20g of poria cocos and 15g of mirabilite for later use;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria into coarse powder, soaking with 6 times of 70% ethanol for 72 hr according to 0120 of 2020 th four-part general rule of Chinese pharmacopeia, stirring during soaking, collecting supernatant, adding 3 times of 70% ethanol, soaking for 48 hr, collecting supernatant, mixing soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 15ml glycerol into the filtrate, stirring, dissolving, adding 70% ethanol, adjusting 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 pericarpium citri reticulatae viride, 25g of poria cocos and 30g of mirabilite for later use;
(2) Crushing radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos into coarse powder, soaking for 120 hours by using 30% ethanol in an amount which is 5 times of the amount of the coarse powder according to a soaking method in the four-part general rule 0120 of 2020 edition of Chinese pharmacopoeia, stirring in the soaking process, pouring supernatant, adding 30% ethanol in an amount which is 4 times of the amount of the coarse powder for soaking for 72 hours, pouring supernatant, combining soaking liquid, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol 20ml into the filtrate, stirring, dissolving, adding 30% ethanol, adjusting 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 pericarpium citri reticulatae viride, 30g of poria cocos and 10g of mirabilite for later use;
(2) Crushing radix ranunculi ternati, ligusticum wallichii, radix bupleuri, blumea balsamifera, pericarpium citri reticulatae viride and poria cocos into coarse powder, soaking for 96 hours by using 5 times of 60% ethanol according to a soaking method under 0120 of the general rule of the four parts of the 2020 edition of Chinese pharmacopoeia, stirring in the soaking process, pouring a supernatant, adding 4 times of 60% ethanol for soaking for 72 hours, pouring the supernatant, combining soaking liquid, and filtering to obtain a filtrate;
(3) Adding Natrii sulfas and glycerol 50ml into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting 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 pericarpium citri reticulatae viride, 20g of poria cocos and 20g of mirabilite for later use;
(2) Pulverizing radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride and Poria into coarse powder, soaking with 6 times of 60% ethanol for 72 hr according to 0120 of the four-part general rule of 2020 edition of Chinese pharmacopeia, stirring during soaking, collecting supernatant, adding 3 times of 60% ethanol, soaking for 72 hr, collecting supernatant, mixing soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and 40ml glycerol into the filtrate, stirring, dissolving, adding 60% ethanol, adjusting to 1000ml, mixing, standing, and filtering.
The products of the examples in this application were tested by the toxicity testing center of the university of medical, guizhou. Physical and chemical index inspection results: mercury (Hg) is less than 1mg/L, arsenic (As) is less than 2mg/L, lead (pb) is less than 10mg/L, cadmium (Cd) is less than 5mg/L, and methanol is less than 2000mg/L, which meets the standard regulation of the national food and drug administration's technical and safety standards for cosmetics (2015 edition); and (3) microorganism index test results: the total number of bacterial colonies (CFU/mL) <10, the mould and the microzyme (CFU/mL) <10, and heat-resistant escherichia coli, pseudomonas aeruginosa and staphylococcus aureus are not detected, and the standard provisions of the technical Specification for cosmetic safety (2015 edition) of the State food and drug administration are met; toxicology test results: acute oral toxicity test results show that LD50 is more than 5000mg/kg, and the oral toxicity test results are practically nontoxic; the results of multiple skin irritation tests show that the integral average value is 0, the skin irritation index is 0-less than 0.5, and the skin irritation strength is nonirritating. Meets the standard regulation of the national food and drug administration' technical Specification for cosmetic safety (2015 edition); the inspection results show that the product meets the national relevant standards and is safe to use.
The applicant carries out research on the action and mechanism of resisting hyperplasia of mammary glands aiming at the traditional Chinese medicine composition obtained in each embodiment in the application, and experimental results prove that the traditional Chinese medicine composition obtained in each embodiment has the effects of preventing and treating hyperplasia of mammary glands of rats in mammary glands models, regulating the levels of hormones E2, LH, PRL and PROG, increasing SOD activity, reducing MDA content and achieving the effect of preventing and treating hyperplasia of mammary glands by activating a Keap1-Nrf2/HO-1 signal channel.
The applicant discloses a specific scheme for the anti-hyperplasia of mammary glands and mechanism research in example 1, which specifically comprises the following steps:
EXAMPLE 1 action and mechanism study of anti-hyperplasia of mammary glands in the Chinese medicinal composition for treating hyperplasia of mammary glands
1 materials and methods
1.1 test drugs
The application of the Chinese medicinal composition for treating hyperplasia of mammary glands comprises the following steps: the external preparation of the Chinese medicinal composition prepared by the method of example 1; mammary lump removing tablet (Lot: 200110): liaoning Nurse (group) Limited for good drug delivery; estradiol benzoate (Lot #: C11924650), progesterone (Lot #: C12082401): shanghai Michelin Biochemical technology, inc.; 4% paraformaldehyde (Lot: 0708A 22): beijing Lei Gen Biotechnology, inc.; 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: solebao Biotechnology Ltd; PVDF film: millipore Trading co.ltd; mouse monoclonal antibody Keap1 (ab 119403), rabbit polyclonal antibody Nrf2 (ab 156883), and rabbit polyclonal antibody HO-1 (ab 137749).
1.2 instruments
A vernier caliper: shanghai Hui one Chili company Limited; leicaTP1020 auto stop extractor: leica embedding machine: leica cm2245 rotary paraffin slicer: an optical microscope: leica, germany; an electronic balance: sidoolis scientific instruments (Beijing) Inc.; -80 ℃ low temperature refrigerator: sanyo corporation of Japan; milli QA pure water treater: millipo tracing Co.ltd, france; W-80A vortex mixer: medical instrument factories of kylin of sea of Jiangsu; an enzyme-labeling instrument: BIO-RAD, USA; oven: shanghai sperm macro laboratory Equipment Co., ltd; a centrifuge: eppendorf, germany.
1.3 Experimental animals
Female Sprague Dawley (SD) rats purchased from beijing huafukang biotech, inc (license number SCXK (jing) 2019-0008).
1.4 grouping and administration
1.5 detection index
24h after the last dose, (1) the rat's 2 nd pair of breast diameter and nipple height were measured precisely with a vernier caliper. (2) Hematoxylin-eosin (HE) staining: the rats were anesthetized by intraperitoneal injection with 7% chloral hydrate (0.5 mL/100 g), and after syncope, the rats were quickly fixed on a rat plate in a supine position, and then the mammary glands of the udder of the rat 2 nd pair were fixed with 4% paraformaldehyde for 2d, subjected to tissue dehydration, paraffin embedding, and finally sectioned for HE staining. And 3, taking the 3 rd and the 4 th mammary glands of the breasts to carry out biochemical index, ELISA and WB detection.
2 results of the experiment
2.1 Effect of the Chinese medicinal composition for treating hyperplasia of mammary glands on weight change of rats
The weight of the rats is increased, the weight of the rats in the blank control group is increased rapidly, the weight of the rats in other building groups is increased slowly, and the weight of the nodules of breast and the liniment group and the weight of each dosage of the liniment group are increased obviously after 21 days of administration (figure 1).
2.2 effects of the Chinese medicinal composition for treating mammary gland hyperplasia 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 statistical significance is achieved (P < 0.05). Compared with a model control group, the mammary nodule elimination tablet group, the traditional Chinese medicine composition for treating mammary gland hyperplasia and the high-dose group can obviously reduce the nipple height and the mammary gland diameter of a mammary gland hyperplasia rat caused by estradiol benzoate and progesterone, and have statistical significance (P < 0.05, see Table 1).
TABLE 1 Effect of the Chinese medicinal composition for treating mammary gland hyperplasia of the present application on the height and diameter of mammary gland hyperplasia rat
Compared with the K group, the method has the advantages that, * P﹤0.05； ** p < 0.01; in comparison with the group C, # P﹤0.05； ## p < 0.01. Note: k: blank control group; c: a model group; r: mammary lump eliminating tablet; l: the low-dose group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands; m: the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application comprises a dosage group; h: the application relates to a traditional Chinese medicine composition high-dose group for treating hyperplasia of mammary glands.
2.3 the influence of the traditional Chinese medicine composition for treating hyperplasia of mammary glands on the pathological forms of the mammary gland tissues
Acinus and lobule of gland of mammary gland tissue of a blank group of rats are normal; the number of mammary gland lobules of a model group rat is increased obviously, a large amount of secretion appears when the glandular cavity of a mammary gland expands, and epithelial cells of a duct are arranged disorderly and are obviously proliferated; compared with a model group, the positive medicine group, the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application and the administration group for intragastric administration have the advantages that the hyperplasia condition of mammary tissues is improved, the number of acini is obviously reduced, the volume of lobules of the gland is obviously reduced, and the duct and lumen secretion of mammary glands is in a reduction trend.
The experimental result shows that compared with a model group, the quality of rats of the positive control group and the Miao medicament of the traditional Chinese medicine composition for treating hyperplasia of mammary glands in each dosage of the application group 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 the mammary tissue is obviously improved.
2.4 the effect of the traditional Chinese medicine composition for treating hyperplasia of mammary glands on MDA content and SOD activity in mammary gland tissues and serum of mammary gland hyperplasia rats
Estradiol benzoate and progesterone induced mammary gland hyperplasia can cause oxidative damage, and SOD activity and MDA content in serum and mammary gland tissues are detected to evaluate the protective effect of the traditional Chinese medicine composition for treating mammary gland hyperplasia on mammary gland hyperplasia oxidative damage. As shown in fig. 3, the SOD activity in mammary tissue and serum of the rats of the model group (C) was significantly decreased, while the content of MDA was significantly increased (P < 0.01) compared to the blank control group (K); after administration, SOD activity and MDA content in rat mammary gland tissues and blood serum of the traditional Chinese medicine composition for treating mammary gland hyperplasia have obvious improvement effect.
2.5 Effect of the Chinese medicinal composition for treating hyperplasia of mammary glands of the application on the level of hormone in mammary gland tissue and serum of mammary gland hyperplasia rats
As shown in FIGS. 4 and 5, the levels of E2, LH and PRL in mammary tissue and serum of the model group (C) rats were significantly increased (P < 0.05) and the level of PROG was significantly decreased (P < 0.05) compared to the blank control group (K); compared with a model group, after the administration of the mammary lump-removing tablet and the traditional Chinese medicine composition for treating hyperplasia of mammary glands, the E2, LH and PRL levels in mammary gland tissues and blood serum of rats are obviously reduced (P is less than 0.05), and the PROG level is obviously increased (P is less than 0.05).
2.6 the Chinese medicinal composition for treating hyperplasia of mammary glands of the application can be used for treating the expression levels of Nrf2, keap1 and HO-1 proteins in mammary gland tissues of mammary gland hyperplasia rats
The results of immunoblotting detection of protein expression of Nrf2, keap1 and HO-1 in mammary tissue of each group of rats are shown in FIG. 6, compared with the blank control group (K), the protein expression of Nrf2 and HO-1 in mammary tissue of rat in the model group (C) is obviously reduced (P is less than 0.01), and the protein expression of Keap1 is obviously increased (P is less than 0.01); compared with a model group, after the administration of the nodules of breast tablet and the traditional Chinese medicine composition for treating hyperplasia of mammary glands disclosed by the application, the protein expression of rat mammary tissue Nrf2 and HO-1 is remarkably increased (P is less than 0.01), and the protein expression of Keap1 is remarkably reduced (P is less than 0.01).
Discussion of 3
The hyperplasia of mammary glands is caused by endocrine disorder, and is closely related to hormone disorder in vivo, especially the imbalance of the proportion of female hormone and progestogen. PRL is a prolactin produced by eosinophils in the distal part of the pituitary, and studies have shown that: PRL has the functions of promoting the growth and development of mammary gland and maintaining lactation, and can generate certain inhibition effect on hypothalamus-pituitary-ovary axis. When PRL in mammary gland tissue and blood is abnormally increased, the secretion of PROG is inhibited, and the synthesis of E2 is stimulated, so that the proportion of E2/PROG is disordered, the mammary gland tissue is badly stimulated, and the hyperproliferation of the mammary gland tissue is caused. E2 can promote the regeneration of mammary ducts, and the expression of the E2 can inhibit LH secretion of anterior pituitary, thereby inhibiting the continuous growth of the mammary ducts on one hand and also having a certain inhibiting effect on the development of mammary lobules on the other hand. The results of this study show 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 disorder of a mammary gland hyperplasia model rat is improved, and the result of combining mammary gland histopathological section shows that the traditional Chinese medicine composition for treating mammary gland hyperplasia has a protective effect on mammary gland hyperplasia.
In addition, the oxidative stress is also an important factor of mammary gland hyperplasia, after estradiol benzoate and progesterone induce mammary gland hyperplasia, the SOD activity in rat serum and mammary gland tissues is obviously reduced, and the content of MDA is obviously increased. Endogenous oxidative state imbalance can lead to activation of antioxidant defense systems, and a Keap1-Nrf2/HO-1 signal pathway is a defensive transduction pathway which is newly discovered in recent years and resists external oxidative stimuli in organisms. Under normal conditions, nrf2 is combined with cytoskeleton-related protein Keap1 and exists in cytoplasm in a form of dimer, when the cytoplasm is stimulated by external oxidative stress factors, nrf2 and Keap1 are dissociated and transferred into cell nucleus, and then the cell nucleus is interacted with an anti-oxidative stress response element AR to induce the expression of proteins such as anti-oxidative protein HO-1 and the like, so that the anti-oxidative effect is further exerted. Westernblot detects the content of Nrf2, keap1 and HO-1 proteins in mammary tissues, and the results show that compared with a blank control group, the protein expression level of Nrf2 and HO-1 in mammary tissues of a model group is remarkably reduced, and the protein expression level of Keap1 is increased (P < 0.01); compared with a model group, each dosage administration group of the traditional Chinese medicine composition for treating hyperplasia of mammary glands can obviously increase the protein expression amount of Nrf2 and HO-1 and reduce the protein expression amount of Keap1 (P < 0.01). The traditional Chinese medicine composition for treating hyperplasia of mammary glands has the function of preventing and treating hyperplasia of mammary glands possibly through the antioxidation mediated by activating the Keap1-Nrf2/HO-1 signal channel.
4 conclusion
The traditional Chinese medicine composition for treating hyperplasia of mammary glands has a good curative effect on a mammary gland hyperplasia model rat, the possible mechanism of the traditional Chinese medicine composition can regulate the levels of hormones E2, LH, PRL and PROG, the SOD activity is increased, the MDA content is reduced, and the effect of preventing and treating hyperplasia of mammary glands is achieved by activating a Keap1-Nrf2/HO-1 signal channel.
Finally, it should be noted that the above embodiments are merely representative examples of the present invention. Obviously, the technical solution of the present invention is not limited to the above-described embodiments, and many variations are possible. All modifications which can be derived or suggested by the person skilled in the art from the present disclosure are to be considered within the scope of the present invention.
Claims (10)
1. A traditional Chinese medicine composition for treating hyperplasia of mammary glands is characterized by comprising the following raw material components: radix Ranunculi Ternati, rhizoma Ligustici Chuanxiong, bupleuri radix, herba Blumeae Balsamiferae, pericarpium Citri Reticulatae viride, poria, and Natrii sulfas.
2. The Chinese medicinal composition for treating mammary gland hyperplasia according to claim 1, wherein the ternate buttercup root, the ligusticum wallichii, the radix bupleuri, the blumea balsamifera, the green tangerine peel and the poria cocos are ethanol extracts of the ternate buttercup root, the ligusticum wallichii, the radix bupleuri, the blumea balsamifera, the green tangerine peel and the poria cocos.
3. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 2, wherein the ethanol extract of ternate buttercup root, ligusticum wallichii, radix bupleuri, blumea balsamifera, green tangerine peel and poria cocos is extracted by the following method: pulverizing the raw materials into coarse powder, soaking in 5-6 times of 30-80% ethanol for 48-120 hr while stirring, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing soaking solutions, and filtering to obtain filtrate.
4. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 1, wherein 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 pericarpium citri reticulatae viride, 10-30 parts of poria cocos and 5-30 parts of mirabilite.
5. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 1, wherein a solvent and an auxiliary material are further added to prepare an external preparation.
6. The traditional Chinese medicine composition for treating hyperplasia of mammary glands according to claim 1, wherein the solvent is 30-80% ethanol, and the adjuvant is glycerol.
7. The preparation method of the traditional Chinese medicine composition for treating hyperplasia of mammary glands according to any one of claims 1 to 6, which is characterized by 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 in 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 under stirring, collecting supernatant, adding 4-3 times of 30-80% ethanol, soaking for 24-72 hr, mixing soaking solutions, and filtering to obtain filtrate;
(3) Adding Natrii sulfas and glycerol into the filtrate, stirring, dissolving, adding 30-80% ethanol, adjusting, mixing, standing, and filtering.
8. The method for preparing a Chinese medicinal composition for treating mammary gland hyperplasia according to claim 7, wherein the glycerol accounts for 1-5% of the total medicinal solution.
9. The method for preparing a Chinese medicinal composition for treating mammary gland hyperplasia according to claim 7, wherein the final liquid medicine is 0.06g-0.2g of the raw material herbs per ml of the liquid medicine.
10. The use of the Chinese medicinal composition for treating mammary gland hyperplasia according to any one of claims 1 to 6 as/for preparing a medicament for treating mammary gland hyperplasia.
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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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| 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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