CN116327831A - A Chinese medicinal composition with effect of resisting hyperplasia of mammary glands - Google Patents
A Chinese medicinal composition with effect of resisting hyperplasia of mammary glands Download PDFInfo
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- CN116327831A CN116327831A CN202310199130.1A CN202310199130A CN116327831A CN 116327831 A CN116327831 A CN 116327831A CN 202310199130 A CN202310199130 A CN 202310199130A CN 116327831 A CN116327831 A CN 116327831A
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Abstract
The invention provides a traditional Chinese medicine composition with an anti-hyperplasia of mammary glands effect, which comprises the following components: the cat ginseng, the common vine herb and the tailless fruit are used in the following specific parts by weight: 10-20 parts of radix clematidis, 10-20 parts of caulis Erycibes and 20-40 parts of Chinese wampee. Preferably, the weight ratio of the tailless fruits to the catginseng is (2-2.5): 1, and the weight ratio of the catmen to the caulis Erycibes is (1-1.2): 1. The Chinese medicinal composition can be further prepared into pills, paste, tablets and other dosage forms. The traditional Chinese medicine composition can reduce viscosity values of whole blood such as low cut, medium cut, high cut and plasma, has the effects of easing pain, reducing nipple diameter, regulating estrogen secretion level of a body and improving tissue morphology, and has the advantages of few prescription medicines, definite effect and small toxic effect.
Description
Technical Field
The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition with an anti-hyperplasia of mammary glands effect.
Background
Hyperplasia of mammary glands is a benign disease of non-tumor non-inflammatory mammary glands, which is abnormal in normal development and degeneration processes of mammary glands, the incidence rate of the benign disease is in an ascending trend year by year, and the hyperplasia of mammary glands clinically appears as unilateral or bilateral breast pain or lump, is related to emotional changes and menstrual cycles, and is well developed for young and middle-aged women of 25-50 years.
The knowledge of traditional Chinese medicine on the disease, hyperplasia of mammary glands and the like belong to the categories of breast nodules and breast nuclei in traditional Chinese medicine. The traditional Chinese medicine concept extends through all aspects of physiological pathology and dialectical treatment of people. The etiology of the disease is not single, the disease is generated in isolation, the disease is related to imbalance of qi, blood and yin and yang of the human body, and the local pathological changes can influence the overall pathological state of the human body. Nodules of breast mainly include liver depression, and deficiency and excess of the remaining viscera should be considered. Wang Kentang in the "certificate treaty rope" record: the depression of the anger is suppressed, accumulation of spleen qi is eliminated, and liver qi is reversed transversely, so that the heart is hidden. "hyperplasia of mammary glands formation is associated with the liver. Liver qi is ascending and active, qi movement is regulated, blood and body fluids are orderly running, body functions are induced, and viscera and meridians are normal in physiological functions. Liver failing to drain qi, qi movement and ascending and descending are abnormal, liver Yu Heng can not check spleen, spleen qi deficiency, and also can not distribute body fluid, water and dampness are trapped in spleen soil, qi stagnation and dampness obstruction, dampness gathers into phlegm, and qi stagnation and phlegm stagnation in breast become lumps, which is called wood not dredging soil. Spleen deficiency failing to transport and transform, qi failing to promote the flow of body fluids, water-dampness stopping and accumulating, dampness accumulating into phlegm, obstructing qi movement and phlegm-dampness stagnation. Qi is the general purpose of blood, qi stagnation causes blood failure and unsmooth blood circulation, resulting in blood stasis. Liver qi is dredged to coordinate qi-lifting of middle energizer, so liver qi is dredged to promote spleen-earth transportation. Spleen governs transportation and transformation, spleen qi is healthy and transported, food essence is sufficient, qi and blood are generated actively, liver has blood and can be hidden, and liver wood is nourished by spleen earth. Liver is nourished, liver qi is raised, spleen qi is dispersed, lung qi is dispersed, and the whole body is nourished. Liver qi stagnation and spleen depression can result in phlegm and phlegm stasis, and stagnation of milk collaterals can be caused, so that liver qi is soothing and spleen regulating effects are achieved. The liver is the mother, mother and child of the heart, and the five zang organs are mutually exclusive. The heart and liver together with the upper energizer, liver depression turns into fire for a long time, and the fire-heat disturbing the heart-mind causes palpitation and restlessness and tension anxiety. "Su-Wen-Ji-fei-Bing Lun (plain question and evaluation of fever theory of diseases)": the "pulse" pertains to the heart and is located in the cells. The uterus is connected with the heart through the veins. The heart-liver qi is depressed, qi movement is unfavorable, qi is excessive to transform fire, fire heat hurts yin, yin blood is deficient, menstrual blood and sea cannot overflow on time, blood is not pushed, and breast is not nourished to pain. Patients with hyperplasia of mammary glands often have problems such as premenstrual mood changes and abnormal menstruation volume, color and quality. The throat is the portal of the lung, and the lung is not moist, and the throat is unfavorable, and dry throat is uncomfortable, has foreign matters, can not appear as cough, and can not fall down the throat, and is shaped like globus hystericus. This condition is often seen in women of childbearing age and is associated with the smooth regulation of liver qi. Liver qi ascending left and lung qi descending right, the distribution of body fluids in the lung, lung qi diffusing and descending promotes the metabolism of water. Liver qi stagnation, lung failing to disperse and descend, chest distress and discomfort can help generate water dampness and phlegm-fluid retention. The water dampness stops gathering, and the dampness gathers into phlegm, which hinders qi movement, qi stagnation and blood obstruction, and vein stasis, and forms a pathological state of phlegm, water and blood stasis. "surgical medical records Assembly" cloud: although the liver disease is caused by the cloud, it is usually in the kidney. The kidney meridian goes up through the liver and is connected with the milk, the liver and kidney are in the lower energizer, and the liver is dredged and discharged to affect the sealing of kidney essence. The kidney essence is filled and kept away from the liver blood. The "yang is the exterior and yin is the interior", and the functional activities of yang are based on the form of yin. Liver blood deficiency, liver failure to nourish the liver, liver-qi stagnation affecting kidney essence exuberance and disharmony of thoroughfare and conception vessels, form mammary gland diseases.
At present, western medicine treatment mainly comprises medicines such as oral hormone, nonsteroidal medicine and the like, clinically commonly used medicines comprise tamoxifen citrate tablets, toremifene, progestogen and the like, and can relieve pain to a certain extent and clinically achieve a certain curative effect. However, adverse reactions such as: menstrual disorder, vaginal discomfort, nausea, dizziness and the like, and endocrine disorders with different degrees are aggravated after stopping the medicine; the operation treatment has a series of problems of risk, easy recurrence after operation, scar after incision healing, and the like.
The search CN103142902B discloses a traditional Chinese medicine composition for treating hyperplasia of mammary glands, belonging to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10 to 30 portions of angelica, 9 to 27 portions of kudzuvine root, 7.5 to 22.5 portions of dodder, 7.5 to 22.5 portions of cowherb seed, 6 to 18 portions of malt, 6 to 18 portions of kelp, 7.5 to 22.5 portions of rhizoma corydalis and 5 to 15 portions of ricepaper pith.
CN201810495030.2 discloses a traditional Chinese medicine composition for treating hyperplasia of mammary glands, which is characterized by comprising the following raw materials in parts by mass: 20-40 parts of dried orange peel, 20-40 parts of burdock, 10-30 parts of cape jasmine fruit, 10-30 parts of honeysuckle, 20-40 parts of semen trichosanthis, 20-40 parts of fructus forsythiae, 20-40 parts of mother grass, 30-50 parts of pollen, 3-20 parts of radix bupleuri, 5-25 parts of green tangerine peel, 30-50 parts of Japanese pricklyash, 20-40 parts of dandelion and 20-40 parts of white spirit.
The traditional Chinese medicine is mainly a compound, but the traditional Chinese medicine is a classical ancient prescription or the compound for treating hyperplasia of mammary glands according to the prior invention has the common characteristics: the number of medicinal materials is large, part of medicinal materials are rare medicinal materials, the cost is high, the research mechanism is complex, the curative effect is uneven, and the pharmacodynamic mechanism is undefined.
Therefore, how to provide a natural, safe, effective and obvious-action traditional Chinese medicine for resisting hyperplasia of mammary glands, which is one or a small compound combination, is a problem to be solved by the technicians in the field.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to research a traditional Chinese medicine composition for inhibiting hyperplasia of mammary glands.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
a Chinese medicinal composition with effect of resisting hyperplasia of mammary glands comprises the following components: the cat ginseng, the common vine herb and the tailless fruit are used in the following specific parts by weight: 10-20 parts of radix clematidis, 10-20 parts of caulis Erycibes and 20-40 parts of Chinese wampee.
Preferably, the weight ratio of the tailless fruits to the catginseng is (2-2.5): 1, and the weight ratio of the catmen to the caulis Erycibes is (1-1.2): 1. The compatibility of medicines in the proportion range is reasonable, and the curative effect is obvious.
Preferably, the preparation method comprises the following steps: decocting radix Ranunculi Ternati, caulis Erycibes and fructus Canarii albi in water for 2-3 times, each time with 12-14 times of water for 1-3 hr, filtering the decoction with 200-300 mesh filter cloth, mixing decoctions, filtering, concentrating the filtrate to obtain soft extract with relative density of 1.20-1.30 (60deg.C), drying, and pulverizing.
Preferably, the Chinese medicinal composition can be further prepared into pills, paste, tablets and other dosage forms.
The invention is further explained and illustrated below:
radix et rhizoma Rhei, caulis seu radix Kadsurae Longipedunculatae, fructus Pyri vine, oryza Glutinosa, and herba Achillea Wilsonianae. Is a habit of taking medicines in Zhejiang province. The cat ginseng has cold nature, bitter taste, lung and stomach meridian, has the effects of clearing heat and detoxicating, dispelling wind and removing dampness, is mainly used for osteomyelitis, sore abscess, rheumatalgia, liver cirrhosis, jaundice, female leucorrhea, peritoneal fluid accumulation, leprosy and the like, and has unique effect on improving immunity.
The caulis Erycibes is also called Bao Gongteng, spicy caulis Sargentodoxae and is a traditional Chinese medicine for resisting rheumatism in Guangdong and Guangxi folks, has the effects of relieving exterior syndrome, inducing sweat, dispelling wind, eliminating dampness, relaxing tendons, activating collaterals, relieving swelling and pain, and is mainly used for treating rheumatism, rheumatalgia, rheumarthritis, rheumatoid arthritis, ischialgia, hemiplegia, traumatic injury and other symptoms.
The Chinese wampee is taken as a medicine by using dry whole herb, has the effects of stopping bleeding, relieving pain, clearing heat and detoxicating, calming liver and stopping wind, and is clinically used for hypertension, hepatitis and the like.
In the prior art, the antecedents of the cat ginseng, the common vine herb and the Chinese feverfew for inhibiting the hyperplasia of mammary glands are not available, and the traditional Chinese medicine composition with three material compositions is not available.
Compared with the prior art, the invention has the advantages that:
compared with the prior art, the invention discloses a traditional Chinese medicine composition with the function of resisting hyperplasia of mammary glands and a preparation method thereof, wherein the traditional Chinese medicine composition has the advantages of easing pain, reducing viscosity values of whole blood, reducing nipple diameter, regulating serum estrogen content of a body, improving tissue morphology, along with few prescription medicines, definite effect and small toxic effect.
The detailed structure of the present invention is further described below with reference to the accompanying drawings and detailed description.
Drawings
FIG. 1 is a flow chart of a preparation process of an embodiment;
FIG. 2 is a detailed histopathological results of the example, wherein FIG. 2 (a) shows the condition of the mammary gland, HE staining, x 200, normal structure of the glandular cavity of the mammary gland, no hyperplasia of mammary glands, catheter dilatation, hypersecretion, etc. in the blank group (animal No. 1F 04). FIG. 2 (b) shows the proliferation of mammary gland acinus, ductal dilatation and increased milk secretion in model control group (animal No. 2F 06) with HE staining, X200. Fig. 2 (c) shows the low dose group (animal No. 3F 02) of the traditional Chinese medicine composition pill prepared in example 1, the mammary gland, HE staining, x 200, slightly hyperplasia of mammary glands acinus, ductal expansion of mammary glands, and increased milk secretion, and fig. 2 (d) shows the dose group (animal No. 4F 03) of the traditional Chinese medicine composition pill prepared in example 1, the mammary gland, HE staining, x 200, hyperplasia of mammary glands acinus. FIG. 2 (e) shows the high dose group (animal No. 5F 10) mammary gland of the Chinese medicinal composition pill prepared in example 1, HE staining, x 200. The mammary gland blebs slightly proliferate, the mammary duct expands, and the milk secretion increases. FIG. 2 (F) shows the positive control group (animal No. 6F 02) with stained mammary gland, HE, proliferation of mammary gland acinus, expansion of mammary duct, and increased milk secretion.
Detailed Description
Example 1
A Chinese medicinal composition with effect of resisting hyperplasia of mammary glands is prepared by taking 10g of radix Ranunculi Ternati and 10g of caulis Erycibes, decocting with water twice, decocting with 14 times of water for 2 hr, filtering decoction with 200 mesh filter cloth, decocting with 12 times of water for 1 hr, filtering decoction with 200 mesh filter cloth, mixing decoctions, filtering, concentrating filtrate to soft extract with relative density of 1.20-1.30 (60deg.C), drying, pulverizing into fine powder, adding appropriate amount of 50% ethanol, making pill, and drying.
Toxicity test of single administration of Chinese medicinal composition pill prepared in example 1 by oral administration of mice
The study is carried out according to the national GLP specification, and the acute toxicity reaction and death condition of ICR mice after twice oral administration of the traditional Chinese medicine composition pill liquid prepared in the example 1 within 24 hours are observed, so that reference data is provided for repeated administration toxicity tests of the mice.
40 SPF grade ICR mice qualified in quarantine are selected for the test, and the weight of the SPF grade ICR mice is 18.0-20.8 g, and the SPF grade ICR mice are bred in 10mm multiplied by 205mm multiplied by 180mm cages, wherein each cage is 5. Raising according to the environmental condition requirements of international (GB 14125-2010) SPF grade experimental animals, quarantining animals and adapting to the environment for 3 days.
The test observes the acute toxicity test reaction of the traditional Chinese medicine composition pill prepared in the example 1 of ICR mice oral gavage. The ICR mice were selected from 40 animals, each half of which was randomly divided into 2 groups according to body weight, and the groups were a blank group and a pill group of the Chinese medicinal composition prepared in example 1 (119.5 g crude drug/kg), each group was 20 animals. The animals are fasted before the experiment, water is not forbidden for more than 12 hours, then pure water and the traditional Chinese medicine composition pill prepared in the example 1 are respectively administrated by oral and gastric administration according to 40mL/kg, the administration is carried out 2 times a day, the interval between the administration and the administration is 6 hours, poisoning performance and characteristics, toxic reaction occurrence and recovery time, death condition and the like of each group of animals are closely observed and recorded within 0-4 hours after each administration, then the animals are observed for 2 times a day, each time in the morning and afternoon, and the animals are continuously observed for 14 days. Animals were weighed on day 4, day 7, day 10 and day 14 before and after the administration, respectively, and weight changes and death were recorded.
Result calculation and statistics
The data significant digit reduction is performed by rounding. The software used for statistics is SPSS16.0, and the weight is average ± standard deviation
And (3) representing. The comparison between the two groups uses t Test when the normalization and variance alignment are satisfied, and Mann-Whitney Test, P when the data do not satisfy the normalization<0.05 represents a statistically significant, P<0.01 indicates that the difference examined is of very significant significance.
Results and analysis
1. Effects on general activity conditions: no obvious abnormality was seen in the autonomous activity, mental state and eating condition of ICR mice in the blank group and the pellet group of the Chinese medicinal composition prepared in example 1 within 0 to 4 hours after the end of each intragastric administration. The mice in the blank group and the pill group of the traditional Chinese medicine composition prepared in the example 1 have no obvious abnormality in autonomous activity, mental state and eating condition after being continuously observed for 14 days after administration.
2. Symptoms of animal poisoning and death: no animal death was seen throughout the trial.
3. Weight of: animals were weighed on day 4, day 7, day 10 and day 14 after dosing, and the weight of the animals was not statistically different from that of the synchronous blank control group, which indicates that ICR mice were orally administered with the Chinese medicinal composition pellets prepared in example 1 and had no obvious effect on weight gain of the mice, as shown in Table 1, 2 and 3.
4. General anatomical examination results: after the test, all the test animals are killed by isoflurane anesthesia, and the animals are subjected to general anatomic examination, and the surface and section shape, size, texture and the like of each organ (lung, liver, kidney, spleen, heart, brain, skin and the like) are observed visually, so that the examination results show that no obvious abnormal condition is seen on the surface and section of each organ.
5. Conclusion and evaluation
Under the test conditions, mice are orally irrigated with the traditional Chinese medicine composition pill prepared in the example 1, the administration volume is 40mL/kg, the administration is carried out 2 times a day, the interval between the two administrations is 6 hours, the accumulated dose is 119.5g crude drug/kg, the dose is 167 times of the dose of the adult clinical intended kilogram body weight, and the animals do not have relevant toxic reaction and death.
Table 1 effect of the Chinese medicinal composition pellets prepared in example 1 on ICR mouse body weight (n=20,
)
table 2 effect of the pill of the traditional Chinese medicine composition prepared in example 1 on body weight of female ICR mice (n=10,
)
table 3 effect of the Chinese medicinal composition pellets prepared in example 1 on body weight of male ICR mice (n=10,
)
toxicity test of the Chinese medicinal composition pill prepared in example 1 by oral administration to rats for 6 months
The purpose is as follows: the project study is carried out according to the national GLP specification, the repeated administration toxicity test of the traditional Chinese medicine composition pill liquid prepared in example 1 with different doses through oral administration of SD rats is observed, the toxic reaction of the traditional Chinese medicine composition pill prepared in example 1 after repeated administration to SD rats is evaluated, and reference data is provided for clinical administration of the traditional Chinese medicine composition pill prepared in example 1.
The test method comprises the following steps: 120 qualified SD rats are selected, the male and female rats are half, the weight is 170.2-218.5 g,475 multiplied by 350 multiplied by 200mm 3 Raising in cages, wherein 5 cages are used for each cage. Raising according to the environmental condition requirements of international (GB 14125-2010) SPF grade experimental animals, quarantining animals and adapting to the environment for 7 days. The animals were randomly divided into 4 groups of 30 animals each, male and female halves, by sex weight. The test was divided into a blank group and a low, medium and high dose group (5.61, 11.21 and 22.41g crude drug/kg) of the Chinese medicinal composition pill prepared in example 1. The administration was performed by gavage in a volume of 15mL/kg for 6 months (26 weeks). At the end of the dosing period (week 13), at the end of the dosing period (week 26) and at the end of the recovery period (week 30) 40 rats were dissected individually, hermaphroditic, and semi-hermaphroditic as planned. The inspection items include: general clinical observations; body weight and food intake were measured; hematology, blood biochemistry, coagulation and organ coefficient measurement; histopathological examination.
Test results: during the test period, all animals were euthanized as planned, and no animals died during the test.
General clinical observations: during observation, each dosage group of the traditional Chinese medicine composition pill prepared in the example 1 is compared with a blank control group in the same period, and animal appearance signs, behavior activities, secretion of each cavity and general conditions of animals and the like do not have abnormal reactions related to drug toxicity.
Weight of: compared with the blank control group, the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on the weight of rats in each dosage group.
Food intake: compared with the blank control group, the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on the feeding amount of rats in each dosage group.
Hematology examination: compared with the blank control group at the same time, the hematology index of each dosage group of the traditional Chinese medicine composition pill prepared in the example 1 is not changed in a toxicological significance, and the result shows that the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on the hematology index of rats.
Biochemical examination of blood: compared with the blank control group at the same time, the blood biochemical indexes of each dosage group of the traditional Chinese medicine composition pill prepared in the example 1 are not changed in toxicological significance, and the result shows that the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on the blood biochemical indexes of rats.
Coagulation examination: compared with the blank control group at the same time, the coagulation index of each dosage group of the traditional Chinese medicine composition pill prepared in the example 1 is not changed in toxicological significance, and the result shows that the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on the coagulation index of rats.
Organ coefficients: compared with the blank control group at the same time, the traditional Chinese medicine composition pill prepared in the example 1 has no obvious abnormality of organ coefficients of each dosage group, and the result shows that the traditional Chinese medicine composition pill prepared in the example 1 has no obvious influence on organ coefficient indexes of rats.
General anatomic examination and microscopic observations: compared with the blank control group at the same period, the high-dose group of the traditional Chinese medicine composition pill prepared in the final-stage male example 1 has pathological changes of hyperplasia of mammary glands, and has no obvious influence on the recovery period.
Conclusion: under the test conditions, the Chinese medicinal composition pill prepared in example 1 was orally administrated to male SD rats for 6 months, and the dose of toxic reaction (NOAEL) obviously related to the medicament was not found to be 11.21g crude drug/kg (equivalent to 16 times of the dose clinically intended for 70kg adults, 2.5 times of the equivalent dose).
Under the test conditions, the Chinese medicinal composition pill prepared in example 1 was orally administrated to female SD rats for 6 months, and the dose of toxic reaction (NOAEL) obviously related to the medicament was 22.41g crude drug/kg (equivalent to 31 times of the dose clinically intended for 70kg adults, 5.0 times of the equivalent dose).
Table 4 effect of the pill of the Chinese medicinal composition prepared in example 1 on animal weight (g,
)
note that: there was no statistical difference between the total male and female animals in each dose group compared to the blank control group.
Table 5 effect of the Chinese medicinal composition pellets prepared in example 1 on the average food intake of animals (g/day,
)
note that: there was no statistical difference between the total male and female animals in each dose group compared to the blank control group.
TABLE 6 influence of the Chinese medicinal composition pill prepared in example 1 on animal hematology
Note that: in comparison with the blank group, * P<0.05， ** P<0.01。
TABLE 7 influence of the Chinese medicinal composition pill prepared in example 1 on the biochemical index of animal blood
Note that: in comparison with the blank group, * P<0.05， ** P<0.01。
TABLE 8 influence of the Chinese medicinal composition pill prepared in example 1 on the coagulation of animals
Note that: there was no statistical difference between the total male and female animals in each dose group compared to the blank control group.
TABLE 9 statistical table of animal organ coefficients for the Chinese medicinal composition pill prepared in example 1
mg/g)
Note that: in comparison with the blank group, * P<0.05。
pharmacodynamic test of Chinese medicinal composition pill prepared in example 1
Experiment one Effect on mammary gland hyperplasia rat model
1. Purpose of investigation
The experiment selects the traditional Chinese medicine composition pill prepared in the example 1 for constructing a mammary gland hyperplasia rat model, and different doses of the traditional Chinese medicine composition pill are administrated to rats through gastric lavage, so that the influence of the traditional Chinese medicine composition pill prepared in the example 1 on mammary gland hyperplasia, whole blood viscosity and plasma viscosity of the rats is observed, and an experimental basis is provided for clinical application.
2. Experimental materials
2.1 test article (test article)
Name: the Chinese medicinal composition pellets (temporary name) prepared in example 1, content: every 1g extract is equivalent to 1.868g crude drug, and the clinical dosage is as follows: 50g crude drug/day, diameter for clinical use: oral administration, 5g once, 3 times a day, clinical treatment course: 3 months, production unit: hunan New Hui pharmaceutical Co., ltd, provided the units: gu Yitang (Hunan) health technologies Co., ltd.
2.2 control drug name: breast nodule eliminating tablet, specification: 0.36g x 72 tablets/bottle, production unit: one of the pharmaceutical companies in Guangzhou white cloud mountain.
2.3 main reagent names: rat estradiol ELISA kit, rat progesterone ELISA kit, manufacturer, jiangsu Jingmei Biotech Co.
2.4 Experimental animals and Experimental systems
2.4.1 laboratory animals
Variety/strain: SD rats, grade: SPF grade, gender: female, number of animals used: body weight range of 60: 177.7-238.5 g (weight after model assembly), 252.2-282.5 g (weight of normal control group), supply unit: hunan Stokes Selaginella, inc., experimental animals produced license number: SCXK (xiang) 2019-0004, experimental animal use license number: SYXK (Hunan) 2018-0004, test animal quality eligibility number: no.430727211102167031.
2.4.3 animal numbering
When the animals are placed in the room, the animals are kept in separate cages, and female rats are denoted by F. After grouping, animals were numbered 2 times with the numbering convention "group number + sex + individual number".
3. Test method
3.1 design of dosage of test and control
The clinical planned dose of the traditional Chinese medicine composition pill prepared in the example 1 is 50g crude drug/day, according to Wei Wei code "pharmacology experiment methodology", fourth edition, the equivalent dose of rats is 50×0.018/0.2=4.5 g crude drug/kg, the maximum clinical planned dose of 1, 2 and 4 times of the equivalent dose of rats is selected as the dose of low, medium and high dose groups, and the doses are respectively 2.24, 4.48 and 8.96g crude drug/kg.
The clinical simulated dose of the breast nucleus resolving hard mass tablet (positive control) is 4.32 g/day, the clinical simulated dose is converted into the equivalent dose of a rat by a body surface area method to be 4.32 multiplied by 0.018/0.2=0.39 g/kg, and the equivalent dose of the rat which is 2 times of the maximum clinical simulated dose is selected as the administration dose in the test, namely 0.78g crude drug/kg, and the table is shown in Table 10.
Table 10 table of administration dosage of the Chinese medicinal composition pill prepared in example 1
Note that: 1g of extract corresponds to 1.868g of crude drug.
3.2 pharmaceutical formulation
The Chinese medicinal composition pill prepared in example 1: weighing the Chinese medicinal composition pill prepared in example 1, adding pure water, stirring, and making into medicinal liquid of 0.224g crude drug/mL, 0.448g crude drug/mL, and 0.896g crude drug/mL, respectively, 1 time a day, and stirring before use.
Breast nodule eliminating tablet: grinding the components of the skin disease blood pill, weighing the powder, adding pure water, stirring and mixing uniformly to prepare 0.078g/mL liquid medicine, and stirring uniformly before use for 1 time a day.
3.3 route of administration, method of administration: is administered orally. Dosing volume: 10mL/kg. Frequency and time of administration: 1 time/day. Drug administration period: for 28 days.
3.4 modeling grouping and administration
After quarantine, animals were randomly divided into 2 groups according to body weight and nipple diameter, 12 blank control groups, and 58 model control groups. The rats in the blank control group were injected with physiological saline alternately on the left and right hind limbs, and the rats in the model control group were injected with estradiol benzoate (0.5 mg/kg) alternately on the left and right hind limbs daily for 25d, and then with progesterone (4 mg/kg) again daily for 7d. And taking two model rats before post-manufacturing administration, carrying out pathological material taking, and judging whether the model is successful. During the injection of estrogen for molding, the tail is clamped by a mouse clamp, so that the rat is stimulated for 30min, and the molding is finished after 1 time per day. After the modeling is finished, animals in the model group are grouped according to the weight and the nipple size to be a model control group, a low-dose group, a medium-dose group, a high-dose group and a positive control group respectively. After grouping, rats in each group were given a volume of 10mL/kg by initial gavage administration, 1 time a day, for 28 days, and rats in the blank group and the model control group were gavaged with pure water.
3.5 general observations
Animals were weighed before administration, and the appearance signs, behavioral activities, secretions, excretions, dietary conditions, death conditions, and the like of each group of animals were closely observed and recorded after administration.
3.6 detection index
1. Measuring nipple diameter after molding, before administration, during administration, and at the end of administration;
2. after last administration, taking the 2 nd pair of right nipple and lower breast tissues of each group of rats for pathological examination;
3. after the last administration, the hemorheology index (whole blood viscosity, plasma viscosity) of the rats was measured. Serum estrogen and progesterone levels were measured.
3.7 data processing and statistical analysis
Data statistics, experimental data were performed using SPSS16.0 software
The normalization and variance alignment tests were performed when comparing the variance alignment among the groups. If the normalization (P>0.05 Statistical analysis was performed using one-way analysis of variance (ANOVA), and comparative analysis was performed using lsd+dunnet (variance) or Tamhane's T2 (variance) depending on variance conditions. If the normalization (P)<0.05 The Kruskal-Wallis test and the Mann-Whitney test were used for pairwise comparison analysis. The statistical result is bounded by a=0.05, where P<0.05 represents a statistically significant, P<0.01 indicates that the difference examined is of very significant significance.
4 experimental results
(1) Effects on the blood rheology index: compared with the blank control group, the whole blood viscosity of the rats in the model control group is obviously increased (P < 0.01) in low, medium, high and plasma viscosity values; compared with the model control group, the low, medium and high dose groups of the Chinese medicinal composition prepared in the example 1 have low, medium and high cut-off values and blood plasma viscosity values which are obviously or obviously reduced (P <0.05 or P < 0.01), and the positive control group (the breast nucleus resolving hard mass tablet) has low, medium and high cut-off values and blood plasma viscosity values which are obviously or obviously reduced (P <0.05 or P < 0.01), and the results are shown in Table 11.
(2) Effect on nipple diameter: compared with the blank control group, the nipple diameter of the rat in the model control group is obviously increased before, during and at the end of the administration after the molding (P <0.05 or P < 0.01); compared with the model control group, the middle administration period of the traditional Chinese medicine composition pill prepared in the example 1 has obviously reduced nipple diameter (P < 0.05) in the high-dose group at the middle administration period; at the end of administration, the Chinese medicinal composition prepared in example 1 has significantly reduced or even reduced nipple diameters (P <0.05 or P < 0.01) in the low, medium and high dose groups. The results of the mid and end dosing period were shown in Table 12, where the positive control (the nodule-dissipating tablet) had significantly or significantly reduced nipple diameter (P <0.05 or P < 0.01) compared to the model control.
(3) Effects on serum estrogen content: compared with the blank control group, the serum estradiol content of the rats in the model control group is obviously increased (P < 0.05), and the progesterone content is obviously reduced (P < 0.01); compared with the model control group, the serum estradiol of the high-dose group and the positive control group in the traditional Chinese medicine composition pill prepared in the example 1 is obviously or obviously reduced (P <0.05 or P < 0.01), and the serum estrogen content of the low-dose group in the traditional Chinese medicine composition pill prepared in the example 1 is reduced, but no statistical difference is seen; the progesterone content of the traditional Chinese medicine composition pill prepared in the example 1, the progesterone content of the high-dose group and the positive control group is obviously or obviously increased (P <0.05 or P < 0.01), the progesterone content of the traditional Chinese medicine composition pill prepared in the example 1, the progesterone content of the low-dose group is increased, but no statistical difference is found, and the result is shown in Table 13.
(4) Histopathological results: the mammary gland of individual animals in the blank control group has hyperplasia of acinus and increased milk secretion, and the mammary gland of other animals has complete structure, no hyperplasia, inflammatory cell infiltration and other pathological changes, and the average score is 0.5; the mammary gland of the animal in the model control group has hyperplasia of acinus, the duct is expanded, the milk secretion is increased, and the average value is 2.9; the various groups of medicines can be seen to have hyperplasia of acinus with different degrees, the catheter is expanded, the milk secretion is increased, and the average of the low, medium and high dosage groups and the positive control group of the traditional Chinese medicine composition pill prepared in the example 1 are 2.1, 1.9 and 2.1 in sequence. The detailed histopathological results are shown in Table 14 and FIG. 2. The mammary gland rehabilitation pill has better treatment effect on SD rat mammary gland hyperplasia model.
TABLE 11 influence of the Chinese medicinal composition pill prepared in example 1 on the blood rheology of rats
n＝10)/>
Note that: compared with the blank control group, "x" indicates P <0.01; in comparison to the model control group, "Δ" means P <0.05, and "ΔΔ" means P <0.01.
TABLE 12 influence of the Chinese medicinal composition pills prepared in example 1 on nipple diameter
n＝10)
Note that: in comparison to the normal control group, "x" indicates P <0.01; in comparison to the model control group, "Δ" means P <0.05, and "ΔΔ" means P <0.01.
TABLE 13 influence of the Chinese medicinal composition pills prepared in example 1 on serum estrogen content
n＝10)
Note that: in comparison to the normal control group, "x" means P <0.05, "x" means P <0.01; in comparison to the model control group, "Δ" means P <0.05, and "ΔΔ" means P <0.01.
Table 14 record of the mammography observation (histopathological results)
Conclusion of the experiment
The experimental results show that: under the test condition, the traditional Chinese medicine composition pills prepared in the example 1 with different doses (2.24, 4.48 and 8.96g crude drugs/kg) for SD rats are used for lavaging, and the low, medium and high dose groups and the positive control group (the breast nucleus powder tablet) of the traditional Chinese medicine composition pills prepared in the example 1 can reduce the viscosity values of whole blood such as low cut, medium cut, high cut and plasma viscosity; can reduce the nipple diameter of a mammary gland hyperplasia rat model; the traditional Chinese medicine composition pill prepared in the embodiment 1, the high dose group and the positive control group (the breast nucleus resolving hard mass tablet) can reduce the serum estradiol content of the mammary gland hyperplasia rat model. The combination of histopathological results shows that the traditional Chinese medicine composition pill prepared in the embodiment 1 has obvious effects of activating blood circulation to dissipate blood stasis and relieving hyperplasia of mammary glands on a mammary gland hyperplasia rat model.
Experiment two analgesic test
1. Purpose of investigation
In the experiment, the traditional Chinese medicine composition pills prepared in the example 1 with different dosages are administrated to mice through stomach irrigation, and the influence of the traditional Chinese medicine composition pills prepared in the example 1 on the analgesic effect of the mice is observed through a hot plate method and an acetic acid torsion method respectively, so that an experimental basis is provided for clinical application.
2. Experimental system
3.1 test article (test article)
Name: the Chinese medicinal composition pellets (temporary name) prepared in example 1, content: every 1g extract is equivalent to 1.868g crude drug, and the clinical dosage is as follows: 50g crude drug/day, diameter for clinical use: oral administration, 5g once, 3 times a day, clinical treatment course: 3 months, production unit: hunan New Hui pharmaceutical Co., ltd, provided the units: gu Yitang (Hunan) health science and technology Co., ltd
2.2 control drug name: aspirin enteric-coated tablet, specification: 500mg×100 tablets/bottle, production unit: shu Taishen (Beijing) biopharmaceutical Co., ltd
2.3 main reagent names: acetic acid, specification: 500 g/bottle, production unit: national medicine group chemical reagent Co., ltd
2.4 Experimental animals and Experimental systems
2.4.1 laboratory animals
Variety/strain: ICR mice, grade: SPF grade, gender: female, number of animals used: 50 (acetic acid torque), 50 (hot plate method), weight range: 21.0 to 25.7g (acetic acid torque), 18.4 to 25.7g (hot plate method), supply unit: hunan Stokes Selaginella, inc., experimental animals produced license number: SCXK (xiang) 2019-0004 laboratory animal use license number: SYXK (Hunan) 2018-0004, test animal quality eligibility number: no.430727211102525624
2.4.3 animal numbering
Animals are kept in separate cages when they enter the room. Female mice are denoted by F. After grouping, animals were numbered 2 times with the numbering convention "group number + sex + individual number".
3. Test method
3.1 design of dosage of test and control
The clinical planned dose of the traditional Chinese medicine composition pill prepared in the example 1 is 50g crude drug/day, according to Wei Wei code "pharmacology experiment methodology", fourth edition, the equivalent dose of mice is 50×0.0026/0.02=6.5 g crude drug/kg, the maximum clinical planned dose of 1, 2 and 4 times of the equivalent dose of rats is selected as the dose of low, medium and high dose groups, and the doses are respectively 3.36, 6.35 and 13.07g crude drug/kg.
The clinical dosage of the aspartame enteric coated tablet (positive control) is 1800 mg/day, the equivalent dose of the aspartame enteric coated tablet is 1800 multiplied by 0.0026/0.02=234 mg/kg according to the body surface area method, and about 1 time of the equivalent dose of the mice is selected as the administration dose of the positive control group, namely 234mg/kg in the test. See table 15.
Table 15 table of administration dosage of the Chinese medicinal composition pill prepared in example 1
3.2 pharmaceutical formulation
The Chinese medicinal composition pill prepared in example 1: weighing the Chinese medicinal composition pill prepared in example 1, adding pure water, stirring, and respectively preparing into medicinal liquid of 0.168g crude drug/mL, 0.318g crude drug/mL, and 0.654g crude drug/mL, 1 time a day, and stirring before use.
Aspirin enteric-coated tablet: grinding aspirin enteric coated tablet into powder, adding 0.5% CMC-Na, suspending to obtain 117mg/mL medicinal liquid, and stirring for 1 time a day.
3.3 routes and methods of administration
Route of administration: is administered orally. Dosing volume: 20mL/kg. Frequency and time of administration: 1 time/day. Drug administration period: 7 days.
3.4 modeling grouping and administration
(1) Hot plate method: before grouping, the pain threshold of each mouse is measured, the time from the self-feeding of the mice into the hot plate to the occurrence of licking of the feet is recorded as the pain threshold, the total measurement is carried out 2 times, jumping-lovers are removed, and the measurement interval is at least 5min each time. Mice with average pain threshold values of 5-30s were selected for subsequent experiments. The qualified mice were randomly divided into a blank group, a low, medium and high dose group (3.36, 6.35, 13.07g crude drug/kg) and a positive control group (aspirin, 234 mg/kg) according to body weight, 10 mice per group. The corresponding dosage of the medicine is administrated by the stomach irrigation according to 20mL/kg, and the blank control group is administrated by the stomach irrigation of equal volume of pure water 1 time a day for 7 days.
(2) Acetic acid torsion body: 50 ICR mice are selected and randomly divided into a blank control group, a low dose group, a medium dose group and a high dose group (3.36, 6.35 and 13.07g crude drug/kg) according to weight layering, 10 mice in each group are respectively subjected to gastric administration of corresponding doses of liquid medicine according to 20mL/kg, 1 time/day and 7 continuous days, and the blank control group is subjected to gastric administration of equal volumes of pure water.
3.5 general observations
Animals were weighed before administration, and the appearance signs, behavioral activities, secretions, excretions, dietary conditions, death conditions, and the like of each group of animals were closely observed and recorded after administration.
3.6 detection index
(1) 30min, 1h, 2h after the last dose, each group of mice was placed in a hotplate instrument to measure pain threshold.
(2) 30min after the last administration, each group of mice was injected with 0.6% physiological saline acetate at 0.1mL/10g of the abdominal cavity, and the number of times that the mice had wriggled (abdominal shrinkage indent, extension hind limb, buttock elevation, creep) within 15min after the injection of acetic acid was observed and recorded.
3.7 data processing and statistical analysis
Data statistics, experimental data were performed using SPSS16.0 software
The normalization and variance alignment tests were performed when comparing the variance alignment among the groups. If the normalization (P>0.05 Statistical analysis was performed using one-way analysis of variance (ANOVA), and comparative analysis was performed using lsd+dunnet (variance) or Tamhane's T2 (variance) depending on variance conditions. If the normalization (P)<0.05 The Kruskal-Wallis test and the Mann-Whitney test were used for pairwise comparison analysis. The statistical result is bounded by a=0.05, where P<0.05 represents a statistically significant, P<0.01 indicates that the difference examined is of very significant significance.
4. Test results
(1) Hot plate method: 30min after administration, compared with a blank control group, the pain threshold of mice in the high-dose group and the positive control group (aspirin) of the traditional Chinese medicine composition pill prepared in the example 1 is obviously increased (P < 0.05); 60 minutes after administration, compared with a blank control group, the pain threshold of the mice in the traditional Chinese medicine composition pill prepared in the example 1, the high-dose group and the positive control group (aspirin) is obviously or obviously increased (P <0.05 or P < 0.01); the pain threshold of mice in the high dose group and the positive control group (aspirin) of the traditional Chinese medicine composition pill prepared in the example 1 is obviously or remarkably increased (P <0.05 or P < 0.01) 120min after administration, and the results are shown in Table 16.
(2) Acetic acid twisting method: the results of the example 1 prepared Chinese medicinal composition with low, medium and high dose and positive control (aspirin) mice were significantly or less wriggled (P <0.05 or P < 0.01) at 30min after administration compared with the blank control, and are shown in Table 17.
TABLE 16 influence of the Chinese medicinal composition pill prepared in example 1 on pain threshold before and after administration to mice
n＝10)
Note that: compared to the blank control, "×" indicates P <0.05.
TABLE 17 influence of the Chinese medicinal composition pill prepared in example 1 on the number of writhing times of mice
n＝10)
Note that: in comparison to the blank, "+" means P <0.05, "+" means P <0.01.
5. Conclusion(s)
Under the test condition, ICR mice are subjected to gastric lavage, different doses (3.36 g crude drug/kg, 6.35 g crude drug/kg) of the traditional Chinese medicine composition pill prepared in the example 1, and the low, medium and high dose groups and the aspirin group of the traditional Chinese medicine composition pill prepared in the example 1 can obviously increase the pain threshold of the mice caused by a hot plate method, and can obviously reduce the twisting times of the mice caused by acetic acid, so that the traditional Chinese medicine composition pill prepared in the example 1 has obvious analgesic effects in the hot plate test and the acetic acid twisting test.
Example 2
A Chinese medicinal composition with effect of resisting hyperplasia of mammary glands comprises the following components: the cat ginseng, the common vine herb and the tailless fruit are used in parts by weight: 10 parts of cat ginseng, 9 parts of common vine, and 22 parts of tailless fruits. The preparation method is similar to that of example 1, and the curative effect is similar to that of the traditional Chinese medicine composition of example 1, and is not repeated here.
The foregoing is a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to apply equivalents and modifications to the technical solution and the concept thereof within the scope of the present invention as defined in the appended claims.
Claims (4)
1. A Chinese medicinal composition with the function of resisting hyperplasia of mammary glands is characterized by comprising the following components in part by weight: the cat ginseng, the vine and the tailless fruits comprise the following specific parts by weight: 10-20 parts of radix clematidis, 10-20 parts of caulis Erycibes and 20-40 parts of Chinese wampee.
2. The composition of claim 1, wherein the ratio of parts by weight of the fruit of Chinese wampee to the ginseng of the cat is (2-2.5): 1, and the ratio of parts by weight of the cat to the vine of the chinese wampee is (1-1.2): 1.
3. The Chinese medicinal composition with the function of resisting hyperplasia of mammary glands according to claim 1, wherein the preparation method comprises the following steps of: decocting radix Ranunculi Ternati, caulis Erycibes and fructus Canarii albi in water for 2-3 times, each time with 12-14 times of water for 1-3 hr, filtering the decoction with 200-300 mesh filter cloth, mixing decoctions, filtering, concentrating the filtrate to obtain soft extract with relative density of 1.20-1.30, drying, and pulverizing.
4. The Chinese medicinal composition with the effect of resisting hyperplasia of mammary glands according to claim 1, wherein the Chinese medicinal composition can be further prepared into dosage forms such as pills, ointment, tablets and the like.
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