Traditional Chinese medicine composition for treating hypothyroidism and preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating hypothyroidism and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine compositions. In the traditional Chinese medicine composition, fomes officinalis ames, stigma croci, aloe, resina scammoniae, kalimeris indica, radix aucklandiae, pogonatherum crinitum and rhizoma nardostachyos are taken as main medicines of a formula, asamur, cortex cinnamomi, rhizoma calami, myrrh branches, euphorbia pekinensis resina, common jasminum seeds and champignon are taken as auxiliary medicines of the formula, and vernonia anthelmintica, hypericum perforatum, black pepper and white pepper are taken as correction medicines of the formula. The traditional Chinese medicine composition is clear in compatibility, has the effects of promoting blood circulation to remove blood stasis, eliminating cold to stop pain, relaxing tendons and activating collaterals and tonifying the liver and kidney, can effectively treat hypothyroidism, and is definite in curative effect on improvement of clinical manifestations and laboratory indexes of patients, safe and reasonable.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine composition technology, and particularly relates to a traditional Chinese medicine composition for treating hypothyroidism, its preparation method and application. Background Technology

[0002] Hypothyroidism is a common endocrine disorder caused by various factors such as autoimmune diseases, medications, and surgery, resulting in insufficient thyroid hormone production or impaired thyroid hormone utilization, leading to a low metabolic rate. Hypothyroidism can adversely affect multiple systems in the body. In the reproductive system, it has a significant impact on male patients; approximately 80% of men with hypothyroidism experience decreased libido, and 40%–50% experience varying degrees of erectile dysfunction. This is mainly because most patients with thyroid dysfunction also have hypothalamic-pituitary-gonadal axis dysfunction, which is the main regulatory mechanism for reproductive function. At the cellular level, apoptosis is programmed to occur by effector genes, and the expression of effector genes is closely related to the occurrence of apoptosis. Under normal circumstances, testicular apoptosis rarely occurs, which is closely related to the high dose-response expression of Bcl-2 and the low dose-response expression of Fas. However, when the body experiences conditions such as hypothyroidism, related tissue cells may undergo significant apoptosis. In this case, the expression of apoptosis-related genes Bcl-2 and Fas participates in regulating the balance and development of related tissue cells. Fas, as one of the main transmembrane substances, can cooperate with corresponding ligands to exert its effects, ultimately leading to apoptosis. Studies have also found that Fas is expressed in related tissue cells. On the one hand, thyroid-stimulating hormone (TSH) can inhibit Fas-related expression; on the other hand, TSH can stimulate related cell proliferation, presumably because it blocks the corresponding apoptosis pathway, leading to an increase in the number of related cells. Simultaneously, the Bcl-2 gene has an effect of inhibiting apoptosis in related tissues; its high expression can prevent the occurrence and development of apoptosis. For example, under normal conditions, Bcl-2 is expressed on epithelial cells within glandular follicles, mainly as cytoplasmic material. However, when thyroid function is abnormal, this balance is disrupted.

[0003] Currently, clinical treatments for hypothyroidism mainly include replacement therapy, drug therapy, and surgery. Replacement therapy generally uses medications such as levothyroxine to supplement insufficient thyroid hormones and restore normal metabolic levels. However, long-term use of replacement therapy may lead to hyperthyroidism, drug dependence, and other problems. Some patients may also experience allergic reactions, palpitations, insomnia, and other discomfort. Regarding drug therapy, commonly used antithyroid drugs such as azathioprine and methimazole can control the symptoms of hypothyroidism to some extent, but long-term use may cause side effects such as liver damage and leukopenia, and some patients may develop drug resistance, leading to unsatisfactory treatment results. Surgical treatment is usually suitable for patients with severe conditions that cannot be controlled by other treatments. It restores normal thyroid function by removing the diseased thyroid tissue or adenoma. However, surgical treatment carries certain risks, and postoperative complications such as bleeding, infection, and vocal cord paralysis may occur.

[0004] It is evident that existing treatments for hypothyroidism have many limitations, necessitating the development of a safe, effective treatment method or drug with minimal side effects. Traditional Chinese medicine (TCM) possesses unique advantages and rich experience in treating endocrine disorders, and its multi-target, holistic regulatory characteristics hold promise for providing a new approach to hypothyroidism treatment. Therefore, developing a TCM composition capable of improving thyroid function has significant practical implications and broad application prospects. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a traditional Chinese medicine composition that has the effects of promoting blood circulation and removing blood stasis, dispelling cold and relieving pain, relaxing muscles and tendons, and tonifying the liver and kidneys. It can effectively treat hypothyroidism and has a definite curative effect and is safe and reasonable in improving patients' clinical manifestations and laboratory indicators.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] A traditional Chinese medicine composition, by weight, comprises the following raw materials: 160-200 parts of Ali Red, 120-150 parts of Aloe, 50-60 parts of Asarum, 75-90 parts of Cinnamon, 50-60 parts of Scammonia resin, 30-40 parts of Camazhi, 30-40 parts of Costus root, 30-40 parts of Cuscuta, 30-40 parts of Nardostachys, 20-30 parts of Saffron, 20-30 parts of Acorus calamus, 1-3 parts of Lagerstroemia indica, 15-20 parts of Myrrh branch, 15-20 parts of Euphorbia pekinensis resin, 15-20 parts of Black pepper, 15-20 parts of White pepper, 15-20 parts of Hypericum perforatum, 15-20 parts of Chrysanthemum indicum, and 25-35 parts of Myrrh seeds.

[0008] Preferably, by weight, the ingredients include the following: 170-190 parts of Ali Red, 130-140 parts of Aloe, 52-56 parts of Asarum, 78-84 parts of Cinnamon, 52-56 parts of Scammonia resin, 34-38 parts of Camazhi, 34-38 parts of Costus root, 34-38 parts of Dodder, 32-35 parts of Nardostachys jatamansi, 25-28 parts of Saffron, 25-28 parts of Acorus calamus, 1.5-2.5 parts of Lagerstroemia indica, 17-19 parts of Myrrh branch, 17-19 parts of Euphorbia pekinensis resin, 17-19 parts of Black pepper, 17-19 parts of White pepper, 17-19 parts of Hypericum perforatum, 17-19 parts of Chrysanthemum indicum, and 28-32 parts of Myrrh seeds.

[0009] Preferably, by weight, the ingredients include the following: 180 parts of Ali Red, 135 parts of Aloe Vera, 54 parts of Asarum, 81 parts of Cinnamon, 54 parts of Scammonia resin, 36 parts of Kamazzhiyus, 36 parts of Costus Root, 36 parts of Dodder, 33 parts of Nardostachys, 27 parts of Saffron, 27 parts of Sweet Flag, 2 parts of Lagerstroemia indica, 18 parts of Myrrh Branch, 18 parts of Euphorbia pekinensis resin, 18 parts of Black Pepper, 18 parts of White Pepper, 18 parts of St. John's Wort, 18 parts of Insectivorous Chrysanthemum, and 30 parts of Myrrh Seed.

[0010] Another object of the present invention is to provide a method for preparing the traditional Chinese medicine composition, comprising the following steps: weighing each raw material in proportion, pulverizing them separately and mixing them to obtain mixture 1; mixing mixture 1 with excipients to obtain mixture 2; granulating mixture 2 and drying it.

[0011] Preferably, all raw materials are pulverized to pass through an 80-100 mesh sieve.

[0012] Preferably, the excipients include sucrose, talc, film coating premix, and ethanol with a volume fraction of 95%.

[0013] Another object of the present invention is to provide the application of the aforementioned traditional Chinese medicine composition or the aforementioned preparation method in the preparation of a medicament for treating hypothyroidism.

[0014] Preferably, the traditional Chinese medicine composition increases the levels of FT3, FT4, and DHT in serum and decreases the level of TSH in serum.

[0015] Preferably, the traditional Chinese medicine composition reduces Fas mRNA expression and increases Bcl-2 and Tert mRNA expression.

[0016] Another object of the present invention is to provide a medicament for treating hypothyroidism, wherein the traditional Chinese medicine composition is the sole effective ingredient or one of the effective ingredients of the medicament.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] This invention provides a traditional Chinese medicine composition, in which Alisma, Saffron, Aloe, Scammonia resin, Camaziris, Costus root, Cuscuta, and Nardostachys jatamansi are the main ingredients. These main ingredients play a key role by directly targeting the main symptoms such as limb numbness, paralysis, and limb pain from different aspects, as well as by regulating the brain, heart, liver, and kidneys. Asarum, Cinnamon, Acorus calamus, Myrrh branch, Euphorbia resin, Myrrh seed, and Lagerstroemia indica are the auxiliary ingredients. These auxiliary ingredients assist the main ingredients in better exerting their effects from multiple perspectives, such as dispelling cold, warming and unblocking, removing dampness, and promoting qi circulation, thereby enhancing the therapeutic effect of the formula on the symptoms. Peppered chrysanthemum, Hypericum perforatum, black pepper, and white pepper are the corrective ingredients. These corrective ingredients regulate the stomach and liver, balance the properties of the ingredients, and coordinate the effects of the ingredients, making the entire formula more mild, safe, and effective. This avoids the adverse effects caused by synergistic or excessive effects between the ingredients, allowing the formula to more accurately target the symptoms while reducing the occurrence of adverse reactions. The traditional Chinese medicine composition of this invention has a clear compatibility. The whole formula has the effects of promoting blood circulation and removing blood stasis, dispelling cold and relieving pain, relaxing muscles and tendons, and tonifying the liver and kidneys. It can effectively treat hypothyroidism and has a definite curative effect and is safe and reasonable in improving patients' clinical manifestations and laboratory indicators. Detailed Implementation

[0019] This invention provides a traditional Chinese medicine composition, comprising the following raw materials by weight: 160-200 parts of Ali red, 120-150 parts of aloe vera, 50-60 parts of asarum, 75-90 parts of cinnamon, 50-60 parts of scammonia resin, 30-40 parts of kamaziruyus, 30-40 parts of costus root, 30-40 parts of dodder, 30-40 parts of nard, 20-30 parts of saffron, 20-30 parts of calamus, 1-3 parts of frankincense, 15-20 parts of myrrh branches, 15-20 parts of euphorbia resin, 15-20 parts of black pepper, 15-20 parts of white pepper, 15-20 parts of St. John's wort, 15-20 parts of anthelmintica, and 25-35 parts of fragrant myrrh seeds. Preferably, by weight, the ingredients include the following: 170-190 parts of Ali Red, 130-140 parts of Aloe, 52-56 parts of Asarum, 78-84 parts of Cinnamon, 52-56 parts of Scammonia resin, 34-38 parts of Camazhi, 34-38 parts of Costus root, 34-38 parts of Dodder, 32-35 parts of Nardostachys jatamansi, 25-28 parts of Saffron, 25-28 parts of Acorus calamus, 1.5-2.5 parts of Lagerstroemia indica, 17-19 parts of Myrrh branch, 17-19 parts of Euphorbia pekinensis resin, 17-19 parts of Black pepper, 17-19 parts of White pepper, 17-19 parts of Hypericum perforatum, 17-19 parts of Chrysanthemum indicum, and 28-32 parts of Myrrh seeds. More preferably, by weight, the ingredients include the following: 180 parts of Ali Red, 135 parts of Aloe Vera, 54 parts of Asarum, 81 parts of Cinnamon, 54 parts of Scammonia resin, 36 parts of Camazhizosin, 36 parts of Costus Root, 36 parts of Dodder, 33 parts of Nardostachys, 27 parts of Saffron, 27 parts of Sweet Flag, 2 parts of Lagerstroemia indica, 18 parts of Myrrh Branch, 18 parts of Euphorbia pekinensis resin, 18 parts of Black Pepper, 18 parts of White Pepper, 18 parts of St. John's Wort, 18 parts of Insectivora, and 30 parts of Myrrh Seed. This invention does not specifically limit the source of the above ingredients; commercially available products in the art are acceptable.

[0020] In this invention, the main ingredients are *Alihong* (a type of saffron), saffron, aloe vera, scammonia resin, *Camazoni* (a type of saffron), costus root, dodder, and nard. *Alihong* is the fruiting body of *Alihong*, a plant in the Polyporaceae family, and has a targeted therapeutic effect on cold-type coughs and asthma, and can warm and resolve phlegm. Saffron has the effect of promoting blood circulation and removing blood stasis, and has a clear therapeutic effect on amenorrhea, abdominal masses, and postpartum blood stasis. Aloe vera can clear bile, clear heat and promote bowel movements, eliminate swelling and ulcers, dry dampness and relieve itching, relieve pain, and kill parasites. It has the effect of treating constipation, hemorrhoids, arthritis, eczema, amenorrhea, jaundice, eye diseases, and intestinal parasitic diseases. Scammonia resin is the milky exudate from the root of *Convolvulus arvense*, a plant in the Convolvulaceae family, and has the effects of purging water, breaking up blood stasis and eliminating masses, and killing parasites. It can be used to treat various types of edema, amenorrhea, abdominal masses, abdominal pain due to intestinal parasites, and constipation. Kamaziriyusi has the effects of generating heat and dryness, clearing blockages, promoting urination and menstruation, softening hardness and relieving pain, warming the lungs and relieving cough, removing jaundice and expelling stones, and drying dampness and healing wounds. It is mainly used to treat damp-cold or phlegm-related diseases, such as cold-type urinary retention, irregular menstruation, splenomegaly, chest pain and cough, damp jaundice, kidney stones, bladder stones, abscesses and damp sores. Costus root promotes qi circulation and relieves pain, strengthens the spleen and promotes digestion. It is used to treat abdominal distension and pain, belching, toothache, loss of appetite and indigestion caused by spleen and stomach qi stagnation. Cuscuta, also known as dodder seed, can tonify the liver and kidneys, strengthen essence and reduce urination, calm the fetus, improve eyesight, and stop diarrhea. It is used to treat lower back and knee weakness, impotence, seminal emission, enuresis, frequent urination, threatened abortion, blurred vision, and spleen and kidney deficiency diarrhea caused by liver and kidney deficiency. Nardostachys chinensis promotes qi circulation and relieves pain, relieves stagnation and invigorates the spleen. It can be used to treat abdominal distension and pain, loss of appetite, and toothache. The main ingredients of the formula are saffron, aloe vera, scammonia resin, caramazizoyus, costus root, dodder and nard, which can directly target the main symptoms of numbness, paralysis and limb pain in the limbs from multiple aspects, and play a key role in the main function of soothing the brain, heart, liver and kidneys.

[0021] Asarum, cinnamon, calamus, myrrh branch, euphorbia resin, fragrant myrrh seed, and frankincense are the auxiliary herbs in the formula. Asarum, also known as asafoetida, has the effect of assisting in the treatment of rheumatoid arthritis, helping to dispel cold and dampness and relieve pain. Cinnamon is an important medicine for treating stomach pain, abdominal pain, cold pain in the waist and knees, impotence, infertility due to cold uterus, dysmenorrhea, and amenorrhea caused by damp-cold constitution or abnormal phlegm. Calamus resolves phlegm, opens the orifices, strengthens the spleen and promotes diuresis, detoxifies and kills parasites, and can be used to treat deep-seated damp-cold constitution, coma, forgetfulness, tinnitus, and deafness. Myrrh branch, also known as myrrh, disperses blood stasis, relieves pain, reduces swelling and promotes tissue regeneration, and can be used to treat chest and abdominal pain due to blood stasis, dysmenorrhea, amenorrhea, abdominal masses, traumatic injuries, carbuncles and sores. When used in combination with other herbs, it can effectively enhance the efficacy of other drugs. Euphorbia resin is a resinous secretion from Euphorbia pekinensis, which has the effects of draining water and eliminating phlegm, reducing swelling and dissipating nodules. It can be used to treat edema, abdominal distension, ascites, phlegm accumulation, shortness of breath and cough, difficulty in urination and defecation, carbuncles and boils. Myrrh seeds are the dried, nearly mature fruits of the Myrrh tree. They have the effects of promoting blood circulation and relieving pain, reducing swelling and promoting tissue regeneration, improving limb function, and can be used to treat abdominal pain due to blood stasis, dysmenorrhea, traumatic injuries, carbuncles and boils. Frankincense, also known as frankincense, is pungent and aromatic, bitter and warming. It can promote blood circulation and relieve pain, reduce swelling and promote tissue regeneration, and relax muscles. It can be used for chest pain, stomach pain, dysmenorrhea, amenorrhea, postpartum blood stasis, abdominal pain due to masses, rheumatic pain, muscle spasms, traumatic injuries, carbuncles and boils. Asarum, cinnamon, calamus, myrrh, euphorbia resin, fragrant myrrh seeds, and lavender are used together as auxiliary herbs in the formula. They assist the main herbs in exerting their effects from multiple perspectives, such as dispelling cold, warming and invigorating, removing dampness, and promoting qi circulation, thereby enhancing the therapeutic effect of the formula on the disease.

[0022] The formula consists of anthelmintic herbs including *Hypericum perforatum*, *Hypericum hyacinthum*, black pepper, and white pepper. *Hypericum perforatum* has the effects of clearing abnormal mucus, expelling parasites, removing dampness and reducing swelling, dispelling cold and relieving pain, promoting pigmentation, and restoring skin color. It can be used to treat damp-cold inflammatory swelling, damp-bi syndrome pain, stomach pain, liver disease, vitiligo, and intestinal parasites. *Hypericum hyacinthum* soothes the liver and relieves depression, clears heat and promotes diuresis, reduces swelling and promotes lactation. It can be used for liver qi stagnation, emotional distress, chest tightness, joint swelling and pain, mastitis, and insufficient lactation. In the formula, it can correct the cold nature of other herbs, prevent stomach damage, and protect spleen and stomach function. Black pepper warms the middle jiao, dispels cold, lowers qi, and eliminates phlegm. It is used for stomach cold vomiting, abdominal pain and diarrhea, loss of appetite, and epilepsy with excessive phlegm. In the formula, it can correct the cold nature of other herbs, enhance the warming effect, and make the warming action more coordinated. White pepper warms the middle jiao (spleen and stomach), lowers qi, and eliminates phlegm. It can be used for cold phlegm and food stagnation, upward rushing of cold qi, cold vomiting and dysentery, and cold abdominal pain. In prescriptions, it can play a role in gently correcting coldness. The combination of anthelmintic chrysanthemum, St. John's wort, black pepper, and white pepper acts as a corrective agent in the prescription. By regulating the stomach and liver's properties, balancing the medicinal properties, and coordinating the effects of the drugs, the entire prescription becomes more mild, safe, and effective. This avoids the adverse effects caused by synergistic or excessive interactions between drugs, allowing the prescription to more accurately target the symptoms while reducing the occurrence of adverse reactions.

[0023] The traditional Chinese medicine composition of this invention has a clear compatibility. The whole formula has the effects of promoting blood circulation and removing blood stasis, dispelling cold and relieving pain, relaxing muscles and tendons, and tonifying the liver and kidneys. It can effectively treat hypothyroidism and has a definite curative effect and is safe and reasonable in improving patients' clinical manifestations and laboratory indicators.

[0024] The present invention also provides a method for preparing the traditional Chinese medicine composition, comprising the following steps: weighing each raw material according to a ratio, pulverizing them separately and mixing them to obtain mixture 1; mixing mixture 1 with excipients to obtain mixture 2; granulating mixture 2 and drying it.

[0025] In the preparation method of this invention, it is preferable to pulverize each raw material to pass through an 80-100 mesh sieve, more preferably to pass through a 90 mesh sieve. The excipients preferably include sucrose, talc, a film-coating premix, and 95% ethanol (by volume); preferably, mixture 1 is mixed with sucrose, talc, the film-coating premix, and 95% ethanol at a mass ratio of 839:30:131:30:80 and then granulated. The granulation in this invention is preferably performed using a wet granulation method; after granulation, it is preferably dried at 60-80°C for 6-8 hours. In a specific embodiment of this invention, the film-coating premix is ​​a gastric-soluble film-coating premix manufactured by Beijing Yingmao Pharmaceutical Co., Ltd.

[0026] The present invention also provides the application of the traditional Chinese medicine composition or the preparation method described herein in the preparation of a drug for treating hypothyroidism. The traditional Chinese medicine composition increases the levels of FT3, FT4, and DHT in serum and decreases the level of TSH in serum. The traditional Chinese medicine composition decreases Fas mRNA expression and increases Bcl-2 and Tert mRNA expression.

[0027] The present invention also provides a medicament for treating hypothyroidism, wherein the traditional Chinese medicine composition is the sole effective ingredient or one of the effective ingredients of the medicament.

[0028] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0029] In a specific embodiment of the present invention, the data statistical processing method is as follows: all experimental data are expressed as mean ± standard deviation, P < 0.05 is considered statistically significant, and the statistical software is SPSS version 25.0.

[0030] Example 1

[0031] A traditional Chinese medicine composition, by weight, comprises the following ingredients: 180 parts of Ali Red, 135 parts of Aloe, 54 parts of Asarum, 81 parts of Cinnamon, 54 parts of Scammonia resin, 36 parts of Kamaziriyus, 36 parts of Costus root, 36 parts of Cuscuta, 33 parts of Nardostachys jatamansi, 27 parts of Saffron, 27 parts of Acorus calamus, 2 parts of Lagerstroemia indica, 18 parts of Myrrh branch, 18 parts of Euphorbia pekinensis resin, 18 parts of Black pepper, 18 parts of White pepper, 18 parts of Hypericum perforatum, 18 parts of Chrysanthemum indicum, and 30 parts of Myrrh seeds.

[0032] The preparation method is as follows: Weigh the above raw materials according to the proportion, crush each raw material to pass through a 90-mesh sieve, and mix the crushed raw materials to obtain mixture 1; mix mixture 1 with sucrose, talc, film coating premix and ethanol with a volume fraction of 95% in a mass ratio of 839:30:131:30:80 to obtain mixture 2; granulate mixture 2 by wet method, and dry it at 70℃ for 8 hours after granulation to obtain the traditional Chinese medicine composition.

[0033] Example 2

[0034] A traditional Chinese medicine composition, by weight, comprises the following ingredients: 190 parts of Ali Red, 130 parts of Aloe, 52 parts of Asarum, 83 parts of Cinnamon, 52 parts of Scammonia resin, 34 parts of Kamaziriyus, 34 parts of Costus root, 34 parts of Cuscuta, 35 parts of Nardostachys jatamansi, 25 parts of Saffron, 25 parts of Acorus calamus, 2.5 parts of Lagerstroemia indica, 17 parts of Myrrh branch, 17 parts of Euphorbia pekinensis resin, 17 parts of Black pepper, 17 parts of White pepper, 17 parts of Hypericum perforatum, 17 parts of Chrysanthemum indicum, and 32 parts of Myrrh seeds.

[0035] The preparation method is the same as in Example 1.

[0036] Example 3

[0037] A traditional Chinese medicine composition, by weight, comprises the following ingredients: 195 parts of Ali Red, 125 parts of Aloe, 58 parts of Asarum, 76 parts of Cinnamon, 58 parts of Scammonia resin, 32 parts of Kamaziriyus, 32 parts of Costus root, 32 parts of Cuscuta, 32 parts of Nardostachys jatamansi, 22 parts of Saffron, 22 parts of Acorus calamus, 3 parts of Lagerstroemia indica, 19 parts of Myrrh branch, 19 parts of Euphorbia pekinensis resin, 19 parts of Black pepper, 19 parts of White pepper, 19 parts of Hypericum perforatum, 19 parts of Chrysanthemum indicum, and 26 parts of Myrrh seeds.

[0038] The preparation method is the same as in Example 1.

[0039] Example 4

[0040] A traditional Chinese medicine composition, by weight, comprises the following ingredients: 165 parts of Ali Red, 148 parts of Aloe, 50 parts of Asarum, 86 parts of Cinnamon, 50 parts of Scammonia resin, 40 parts of Kamaziriyus, 39 parts of Costus root, 39 parts of Cuscuta, 38 parts of Nardostachys jatamansi, 28 parts of Saffron, 28 parts of Acorus calamus, 1.5 parts of Lagerstroemia indica, 16 parts of Myrrh branch, 16 parts of Euphorbia pekinensis resin, 16 parts of Black pepper, 16 parts of White pepper, 16 parts of Hypericum perforatum, 16 parts of Chrysanthemum indicum, and 35 parts of Myrrh seeds.

[0041] The preparation method is the same as in Example 1.

[0042] Example 5

[0043] The efficacy of the traditional Chinese medicine composition of this invention in treating hypothyroidism was evaluated using a rat model of hypothyroidism induced by propylthiouracil (PUT).

[0044] The traditional Chinese medicine composition provided in Example 1 of this invention is denoted as BYD.

[0045] 1. Establishment of a rat model of hypothyroidism induced by propylthiouracil (PUT):

[0046] Sixty male SD rats, weighing 250–300 g, were randomly divided into an experimental group (n=50) and a control group (n=10) after the quarantine period. The experimental group received 20 mL / kg of PTU solution (0.05% concentration diluted in distilled water), while the control group received the same volume of distilled water. The model was established for 4 weeks. Before modeling, all rats were in good condition with smooth fur, normal food and water intake, and stable weight gain. In the hypothyroidism model group, after modeling, rats gradually showed a significant decrease in food and water intake, dry and frizzy fur that easily fell out, lethargy, huddling together, and reduced activity. This indicated successful modeling.

[0047] After successful model establishment, the experimental group rats were divided into 5 groups of 10 rats each, and the drug dosages are shown in Table 1:

[0048] Model group: Rats were administered 20 mL / kg of distilled water by gavage daily for 4 consecutive weeks;

[0049] Positive drug group: Rats were given levothyroxine by gavage daily at a dose of 0.94 mg / kg for 4 consecutive weeks;

[0050] BYD low-dose group: Rats were administered the traditional Chinese medicine composition provided in Example 1 by gavage daily. The daily gavage dose was 0.22 g / kg based on the weight of the granulated traditional Chinese medicine composition, and the gavage was continued for 4 weeks.

[0051] BYD medium-dose group: Rats were administered the traditional Chinese medicine composition provided in Example 1 by gavage daily. The daily gavage dose was 0.43 g / kg based on the weight of the granulated traditional Chinese medicine composition, and the gavage was continued for 4 weeks.

[0052] BYD high-dose group: Rats were administered the traditional Chinese medicine composition provided in Example 1 by gavage daily. The daily gavage dose was 0.86 g / kg based on the weight of the granulated traditional Chinese medicine composition, and the gavage was continued for 4 weeks.

[0053] The control group continued to be administered distilled water by gavage, with the same volume as the model group, for 4 consecutive weeks.

[0054] Table 1 Dosage

[0055] Group dose Dosage volume (mL / kg) Dosage concentration % Administration method Number of animals (individuals) control group --- 20.0 --- Gavage 10 Model Group --- 20.0 --- Gavage 10 Euthyrox 0.94 mg / kg 20.0 <![CDATA[4.70×10 -3 ]]> Gavage 10 BYD low-dose group 0.22g / kg 20.0 1.10 Gavage 10 BYD medium-dose group 0.43g / kg 20.0 2.15 Gavage 10 BYD high-dose group 0.86g / kg 20.0 4.30 Gavage 10

[0056] 2. Methods and Observation Indicators

[0057] 2.1 Daily observation: During the experiment, the mental state, coat appearance, activity, feces, and any deaths of the rats were observed daily. The weight data was recorded once a week, and the rectal temperature, 24-hour food intake and water intake were measured until the end of the experiment.

[0058] 2.1.1 Routine observation: After administration, the animals in each group gradually increased their food and water intake, their fur gradually became shinier, shedding decreased, and their mental state improved.

[0059] 2.1.2 The results of the effect on body weight are shown in Table 2. Compared with the control group, the mean body weight of the model group was lower than that of the drug-treated group, and there were statistically significant differences in the mean body weight at each time point (P < 0.01). Compared with the model group, the weight gain of each drug-treated group gradually exceeded that of the model group. On day 28 of drug administration, the mean body weight of the high-dose BYD group was significantly higher than that of the model group (P < 0.01). On days 22 and 28 of drug administration, the mean body weight of the positive control drug group was significantly higher than that of the model group (P < 0.05, P < 0.01).

[0060] Table 2. Effects of traditional Chinese medicine composition on body weight in hypothyroid rats ( n=10)

[0061]

[0062]

[0063] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0064] 2.1.3 The results of the effect on rectal temperature are shown in Table 3. During the administration period, compared with the control group, the rectal temperature of the model group was lower than that of the control group, and the mean weight at each time point was significantly different (P < 0.01). Compared with the model group, although the mean rectal temperature of each administration group was higher than that of the model group, no statistical difference was observed.

[0065] Table 3. Effects of traditional Chinese medicine composition on rectal temperature in hypothyroid rats ( n=10)

[0066] Group dose Day 1 of medication Day 8 of medication Day 15 of medication Day 22 of drug administration Day 28 of drug administration control group --- 37.4±0.4 38.1±0.3 38.1±0.2 37.3±0.4 38.1±0.1 Model Group --- <![CDATA[35.7±0.9 ** ]]> <![CDATA[35.9±0.8 ** ]]> <![CDATA[36.0±0.6 ** ]]> <![CDATA[36.1±1.1 * ]]> <![CDATA[36.8±0.7 ** ]]> Positive drug group 0.94 mg / kg 35.5±0.4 35.9±0.4 35.9±0.3 35.9±0.4 36.7±0.3 BYD low-dose group 0.22g / kg 35.4±0.4 35.8±0.3 35.8±0.3 35.8±0.5 36.6±0.4 BYD medium-dose group 0.43g / kg 35.3±0.3 35.6±0.3 35.7±0.3 35.7±0.6 36.6±0.3 BYD high-dose group 0.86g / kg 35.4±0.5 35.8±0.5 36.0±0.4 35.9±0.7 37.0±0.5

[0067] Note: Compared with the control group, ** P<0.01.

[0068] 2.1.4 The effects of BYD on food intake and water intake are shown in Tables 4 and 5. During the administration period, compared with the control group, the water intake of the model group was lower than that of the control group, and there were significant differences in food intake at each time point (P < 0.05, P < 0.01). Compared with the model group, although the water intake of each administration group fluctuated, no statistical difference was observed.

[0069] Table 4. Effects of traditional Chinese medicine composition on food intake in hypothyroid rats ( n=10)

[0070] Group dose Day 1 of medication Day 8 of medication Day 15 of medication Day 22 of drug administration Day 28 of drug administration control group --- 31.0±0.5 29.7±0.3 27.0±0.2 25.3±0.0 28.6±2.5 Model Group --- <![CDATA[14.7±0.5 ** ]]> <![CDATA[14.6±0.7 ** ]]> <![CDATA[13.1±0.6 ** ]]> <![CDATA[11.8±0.2 ** ]]> <![CDATA[12.7±0.4 * ]]> Positive drug group 0.94 mg / kg 17.0±1.2 16.9±0.5 <![CDATA[17.8±0.3 # ]]> <![CDATA[16.9±0.5 ## ]]> <![CDATA[17.1±0.7 # ]]> BYD low-dose group 0.22g / kg 16.2±3.0 14.1±0.7 14.6±0.3 <![CDATA[13.7±0.0 ## ]]> 13.7±2.0 BYD medium-dose group 0.43g / kg 16.6±0.4 12.4±0.8 14.0±0.3 16.0±2.4 12.9±1.1 BYD high-dose group 0.86g / kg 17.0±1.2 14.9±0.1 16.1±0.4 <![CDATA[20.7±1.1 ## ]]> 17.4±4.2

[0071] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0072] Table 5. Effects of traditional Chinese medicine composition on water intake in hypothyroid rats ( n=10)

[0073] Group dose Day 1 of medication Day 8 of medication Day 15 of medication Day 22 of drug administration Day 28 of drug administration control group --- 37.9±0.5 58.1±2.0 49.5±1.1 47.1±2.4 44.8±4.8 Model Group --- <![CDATA[26.3±1.7 * ]]> <![CDATA[34.3±8.6 ** ]]> <![CDATA[39.1±13.7 ** ]]> <![CDATA[25.3±3.1 * ]]> <![CDATA[25.2±0.6 * ]]> Positive drug group 0.94 mg / kg 33.8±4.0 27.1±4.7 <![CDATA[36.7±10.0 # ]]> 27.6±1.6 30.3±2.4 BYD low-dose group 0.22g / kg 29.9±7.9 25.1±3.5 26.4±5.5 23.4±3.6 24.3±4.6 BYD medium-dose group 0.43g / kg 30.9±3.2 24.8±0.1 28.7±0.5 37.5±1.1 24.5±4.1 BYD high-dose group 0.86g / kg <![CDATA[35.0±0.7 # ]]> 38.5±4.5 44.3±3.2 20.7±4.2 32.1±5.1

[0074] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0075] 2.2 Detection of thyroid hormone levels:

[0076] After the last administration, patients were kept on a fasting diet but allowed free access to water for 12 hours. Blood was collected from the abdominal aorta, centrifuged at 3000 rpm for 10 minutes, and serum was separated. The levels of T4, T3, TSH, and DHT were measured strictly according to the instructions of the ELISA diagnostic kit.

[0077] The results of thyroid hormone level tests are shown in Table 6:

[0078] Compared with the control group, the serum FT3 level in the model group was significantly decreased (P<0.01). Compared with the model group, the serum FT3 level in the high, medium and low dose BYD groups was significantly increased (P<0.05, P<0.01), and the FT3 level in the positive control drug group was also significantly increased.

[0079] Compared with the control group, the serum FT4 level in the model group was significantly decreased (P<0.01). Compared with the model group, the serum FT4 level in the high-dose BYD group and the positive control group was significantly increased (P<0.05, P<0.01).

[0080] Compared with the control group, the serum TSH level in the model group was significantly increased (P < 0.01). Compared with the model group, the serum TSH levels in the BYD medium- and high-dose groups and the positive control group were significantly decreased (P < 0.05, P < 0.01).

[0081] Compared with the control group, the serum DHT level in the model group was significantly lower (P < 0.01). Compared with the model group, the serum DHT levels in the BYD medium- and high-dose groups and the positive control group were significantly higher (P < 0.05, P < 0.01).

[0082] Table 6. Thyroid hormone levels in each group of experimental rats

[0083] Group dose n FT3 (pg / mL) n FT4 (pg / mL) n TSH (ng / mL) n DHT (ng / mL) control group --- 9 38.13±9.26 8 13.23±3.04 10 10.29±2.30 10 4.58±1.37 Model Group --- 9 <![CDATA[20.46±3.04 ** ]]> 9 <![CDATA[4.22±2.59 ** ]]> 10 <![CDATA[16.34±4.02 ** ]]> 10 <![CDATA[1.57±0.47 ** ]]> Positive drug group 0.94 mg / kg 9 <![CDATA[35.45±9.11 ## ]]> 9 <![CDATA[8.55±3.96 ## ]]> 10 <![CDATA[12.80±2.81 # ]]> 10 <![CDATA[2.61±0.80 # ]]> BYD low-dose group 0.22g / kg 9 <![CDATA[28.76±6.79 # ]]> 9 6.44±2.94 10 13.92±4.09 10 2.38±0.83 BYD medium-dose group 0.43g / kg 9 <![CDATA[31.58±9.99 ## ]]> 9 7.05±2.33 10 <![CDATA[11.78±2.82 ## ]]> 10 <![CDATA[2.73±0.91 # ]]> BYD high-dose group 0.86g / kg 9 <![CDATA[32.05±7.93 ## ]]> 9 <![CDATA[7.52±3.01 # ]]> 10 <![CDATA[10.59±3.96 ## ]]> 10 <![CDATA[3.19±1.41 ## ]]>

[0084] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0085] 2.3 The expression of Fas, Bcl-2, and Tert mRNA in the testes was determined using RT-PCR.

[0086] Weigh 100 mg of ultra-low temperature frozen rat testicular tissue and extract total RNA using the TRIZOL method. Follow the kit instructions as follows: (1) Pour the tissue sample ground with liquid nitrogen into a 1.5 mL EP tube containing 500 μL of TransZolUp enzyme-free solution, vortex to mix, add 100 μL of chloroform substitute, mix thoroughly, and let stand for 5 min. (2) Centrifuge the mixture at 4℃, 10000 x g, for 15 min. Carefully aspirate 200 μL of the supernatant into a 1.5 mL EP tube, add 200 μL of isopropanol pre-chilled at 4℃, invert to mix, and incubate at room temperature for 10 min. (3) Centrifuge the solution at 4℃, 10000 x g, for 10 min, carefully discard the liquid, add 0.5 mL of 75% ethanol pre-chilled at 4℃, invert several times to wash the RNA precipitate, centrifuge at 4℃, 7500 x g, for 5 min, and discard the liquid (perform the washing step 2-3 times). (4) Dry at room temperature for several minutes to allow the ethanol to evaporate completely, then add 30 μL of RNA dissolving solution to fully dissolve the RNA. (5) Take 1 μL of RNA and determine the RNA concentration using an ultra-micro spectrophotometer.

[0087] Reverse transcription to synthesize cDNA: Adjust the total RNA concentration of all samples to 500 ng, and perform reverse transcription using a reverse transcription kit. The operation steps are as follows: (1) Prepare the reverse transcription system according to Table 2. (2) Mix gently. (3) Remove at 42℃ for 15 min, then at 85℃ for 5 s to end the reverse transcription experiment. (4) Dilute the cDNA obtained after reverse transcription 5 times and use it as a template for fluorescence quantification. Primer information is shown in Table 7.

[0088] Table 7 Primers and related information

[0089]

[0090] The expression results of Fas, Bcl-2, and Tert mRNA genes in each group of rats are shown in Table 8. Compared with the control group, the expression of Bcl-2 and Tert mRNA in the testicular tissue of the model group rats was decreased, while the expression of Fas was increased (P < 0.05, P < 0.01). Compared with the model group, the expression of Fas mRNA was decreased in the positive control group and the low-dose BYD group (P < 0.05), the expression of Tert mRNA was increased in the medium-dose BYD group (P < 0.05), and the expression of Bcl-2 and Tert mRNA was increased in the high-dose BYD group (P < 0.05, P < 0.01).

[0091] Table 8 Effects of Bcl-2, Tert mRNA, and Fas gene expression in rat testicular tissue.

[0092] Group dose n Bcl-2 n Fas n Tert control group --- 6 1.414±0.227 6 0.846±0.305 6 1.308±0.216 Model Group --- 5 <![CDATA[1.055±0.113 ** ]]> 6 <![CDATA[1.418±0.427 ** ]]> 6 <![CDATA[0.932±0.123 * ]]> Positive drug group 0.94 mg / kg 5 1.075±0.065 6 <![CDATA[0.885±0.367 ## ]]> 6 1.048±0.352 BYD low-dose group 0.22g / kg 6 1.003±0.093 6 <![CDATA[1.019±0.214 # ]]> 6 1.026±0.259 BYD medium-dose group 0.43g / kg 6 1.279±0.197 5 1.297±0.323 6 <![CDATA[1.355±0.341 # ]]> BYD high-dose group 0.86g / kg 6 <![CDATA[1.443±0.263 # ]]> 6 1.244±0.127 6 <![CDATA[1.644±0.396 ## ]]>

[0093] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0094] 2.4 Effects on the expression of Bcl-2, Fas, and FasL proteins in rat testicular tissue

[0095] The operation steps are as follows: (1) Take rat testicular tissue, cut the tissue into small pieces, add lysis buffer at a ratio of 1:10 (add protease inhibitor and protein phosphatase inhibitor at a ratio of 100:1:1 before use) into a special grinding centrifuge tube, grind the tissue with a tissue grinder, centrifuge at 12000 rpm for 15 min at 4℃, and collect the supernatant. (2) Quantify the total protein in the supernatant according to the instructions of the BCA protein quantification kit. (3) Calculate the required volume of PBS and 4× Loading Buffer for dilution based on the final protein concentration of the sample, and add them to a new centrifuge tube according to the calculated amount. Denature the protein by boiling at 100℃ for 10 min in a metal bath, cool it quickly on ice, aliquot and store in a -80℃ freezer. (4) Select the concentration of separating gel and stacking gel according to the molecular weight of the protein to be measured. (5) Load 5 μL of sample into each well. Set the upper gel to a constant voltage of 70V for 45 min. When running the lower gel, adjust the voltage to 110V and stop electrophoresis when the bromophenol blue dye is almost at the end of the gel. This process takes about 60 min. (6) Cut the PVDF membrane to the size of the gel. Activate it in methanol for 30 s and equilibrate it in transfer buffer for 5 min. Perform wet transfer in the "sandwich" mode at a constant voltage of 110V for 60 min. (7) Block with 5% skim milk powder at room temperature for 2 h. (8) Wash the membrane three times with TBST buffer for 10 min each time. (9) Add the diluted primary antibody and incubate overnight at 4°C. (10) Wash the membrane. (11) Add the secondary antibody (1:10000) and incubate at room temperature for 2 h. (12) Wash the membrane. (13) Expose the membrane using ECL luminescent solution.

[0096] The expression results of Bcl-2, Fas, and FasL proteins in rat testicular tissue are shown in Table 9. Compared with the control group, the expression of Bcl-2 and Tert mRNA in the testicular tissue of the model group rats was decreased, while the expression of Fas and FasL was increased (P < 0.05, P < 0.01). Compared with the model group, the expression of Fas and FasL mRNA was decreased in the positive control group and the low-dose BYD group (P < 0.01), the expression of Tert mRNA was increased in the medium-dose BYD group (P < 0.05), and the expression of Bcl-2 and Tert mRNA was increased in the high-dose BYD group (P < 0.05, P < 0.01).

[0097] Table 9. Effects of Bcl-2, Fas, and Fas-L protein expression in rat testicular tissue ( n=3)

[0098] Group dose Bcl-2 / β-Actin Fas / β-Actin FasL / β-Actin control group --- 1.085±0.193 0.488±0.097 0.270±0.024 Model Group --- <![CDATA[0.412±0.091 ** ]]> <![CDATA[1.236±0.143 ** ]]> <![CDATA[1.085±0.070 ** ]]> Positive drug group 0.94 mg / kg 0.376±0.092 <![CDATA[0.432±0.047 ## ]]> <![CDATA[0.312±0.013 ## ]]> BYD low-dose group 0.22g / kg 0.452±0.070 1.208±0.15 1.049±0.092 BYD medium-dose group 0.43g / kg <![CDATA[0.820±0.140 # ]]> <![CDATA[0.797±0.152 ## ]]> <![CDATA[0.686±0.159 ## ]]> BYD high-dose group 0.86g / kg <![CDATA[1.232±0.092 ## ]]> <![CDATA[0.447±0.038 ## ]]> <![CDATA[0.303±0.035 ## ]]>

[0099] Note: Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01.

[0100] The results indicate that after 4 weeks of BYD intervention, the slow weight gain in hypothyroid rats was significantly improved, and food intake and body temperature increased. It also increased serum FT3, FT4, and DHT levels while decreasing TSH levels, thereby alleviating symptoms of hypothyroidism. Furthermore, it reduced Fas mRNA expression, increased Bcl-2 and Tert mRNA expression, increased BCL-2 protein expression, and significantly decreased FAS and FASL protein expression. These results suggest that BYD can treat hypothyroidism by regulating thyroid hormone levels and testosterone levels. Its mechanism of action may be related to regulating the expression of Fas, Bcl-2, and Tert mRNA genes, thereby affecting the expression of FAS, FASL, and BCL-2 proteins.

[0101] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The ingredients, by weight, include the following: 160-200 parts of Ali Red, 120-150 parts of Aloe Vera, 50-60 parts of Asarum, 75-90 parts of Cinnamon, 50-60 parts of Scammonia resin, 30-40 parts of Kamazzhiyus, 30-40 parts of Costus Root, 30-40 parts of Dodder, 30-40 parts of Nardostachys, 20-30 parts of Saffron, 20-30 parts of Sweet Flag, 1-3 parts of Lagerstroemia indica, 15-20 parts of Myrrh Branch, 15-20 parts of Euphorbia pekinensis resin, 15-20 parts of Black Pepper, 15-20 parts of White Pepper, 15-20 parts of St. John's Wort, 15-20 parts of Insectivorous Chrysanthemum, and 25-35 parts of Myrrh Seeds.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients, by weight, include the following: 170-190 parts of Ali Red, 130-140 parts of Aloe Vera, 52-56 parts of Asarum, 78-84 parts of Cinnamon, 52-56 parts of Scammonia resin, 34-38 parts of Kamazzhiyus, 34-38 parts of Costus Root, 34-38 parts of Dodder, 32-35 parts of Nardostachys, 25-28 parts of Saffron, 25-28 parts of Sweet Flag, 1.5-2.5 parts of Lagerstroemia indica, 17-19 parts of Myrrh Branch, 17-19 parts of Euphorbia pekinensis resin, 17-19 parts of Black Pepper, 17-19 parts of White Pepper, 17-19 parts of St. John's Wort, 17-19 parts of Insectivorous Chrysanthemum, and 28-32 parts of Myrrh Seeds.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, By weight, the ingredients include the following: 180 parts Ali Red, 135 parts Aloe, 54 parts Asaron, 81 parts Cinnamon, 54 parts Scammonia resin, 36 parts Kamazzhiyus, 36 parts Costus root, 36 parts Dodder, 33 parts Nardostachys jatamansi, 27 parts Saffron, 27 parts Sweet flag, 2 parts Lagerstroemia indica, 18 parts Myrrh branches, 18 parts Euphorbia resin, 18 parts Black pepper, 18 parts White pepper, 18 parts St. John's wort, 18 parts Insectivora chinensis, and 30 parts Myrrh seeds.

4. The method for preparing the traditional Chinese medicine composition according to claims 1 to 3, characterized in that, The process includes the following steps: weigh each raw material according to the proportion, crush them separately and then mix them to obtain mixture 1; mix mixture 1 with the auxiliary materials to obtain mixture 2; granulate mixture 2 and then dry it.

5. The preparation method according to claim 4, characterized in that, All raw materials are crushed to pass through an 80-100 mesh sieve.

6. The preparation method according to claim 4, characterized in that, The excipients include sucrose, talc, film coating premix, and ethanol with a volume fraction of 95%.

7. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 or the preparation method according to any one of claims 4 to 6 in the preparation of a medicament for treating hypothyroidism.

8. The application according to claim 7, characterized in that, The traditional Chinese medicine composition increases the levels of FT3, FT4, and DHT in serum, and decreases the level of TSH in serum.

9. The application according to claim 7, characterized in that, The traditional Chinese medicine composition reduces Fas mRNA expression and increases Bcl-2 and Tert mRNA expression.

10. A drug for treating hypothyroidism, characterized in that, The traditional Chinese medicine composition according to any one of claims 1 to 3 is the sole effective ingredient or one of the effective ingredients of the drug.

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