Traditional Chinese medicine composition and application thereof

Abstract

The invention discloses a traditional Chinese medicine composition and application thereof, the traditional Chinese medicine composition is prepared from a traditional Chinese medicine extract and a pharmaceutically acceptable carrier, and the traditional Chinese medicine composition is characterized by being prepared from 30-70% of semen cuscutae, 10-50% of fructus lycii, 10-20% of radix rehmanniae recen and 1-10% of radix cyathulae; the invention also discloses application of the traditional Chinese medicine composition in preparation of a medicine for treating infertility.

Description

Technical Field

[0001] The invention relates to a medical use of a traditional Chinese medicine composition, namely, an application of the traditional Chinese medicine composition in preparing an infertility drug. Background Art

[0002] Infertility is a common disease at present. Infertility is divided into infertility syndrome and sterility syndrome. It refers to the failure of successful pregnancy of a couple of childbearing age who have lived together for more than one year, have normal sexual life, and do not take any contraceptive measures. Infertility is for women. Infertility is the failure of pregnancy for one year with normal sexual life without taking any contraceptive measures; sterility is for men, that is, the failure of pregnancy for one year with normal sexual life. The degree of modern infertility has increased significantly and has reached an unprecedented level. It has obviously become a modern disease. Infertility will not only affect physical health, but also bring troubles to the family. The disease will cause family conflicts and destroy happy marriages. For this reason, we have proposed a traditional Chinese medicine for the treatment of infertility to solve this problem.

[0003] Chinese medicine is currently a good choice for infertility treatment. Chinese medicine conditioning has more advantages than other treatment methods, with a shorter course of treatment, lower cost and better effect. Chinese medicine treats infertility, treats based on syndrome differentiation, gives Chinese medicine treatment, strengthens the body's internal environment, regulates the body's internal environment, and brings hormone levels within the normal range, thereby promoting the normal development of follicles, normal ovulation, fertilization and pregnancy.

[0004] Rehmannia glutinosa (Gaert.) Libosch. ex Fisch. et Mey.) is a perennial herb of the genus Rehmannia in the family Scrophulariaceae. It enters the heart, liver, and kidney meridians; it clears away heat and cools blood, nourishes yin, and promotes the production of body fluid; it is used for red tongue, thirst, internal heat due to yin deficiency, bone steaming, fatigue fever, internal heat and thirst, vomiting blood, epistaxis, and spots and rashes.

[0005] Lycium barbarum is the mature fruit of Lycium barbarum, a plant of the Solanaceae family. It is a food and medicine approved by the Ministry of Health. It is sweet, flat, and enters the liver and kidney meridians. It can nourish the liver and kidneys, improve sperm quality and improve eyesight.

[0006] Cuscuta chinensis Lam., an annual parasitic herb of the genus Cuscuta in the family Convolvulaceae, is a good medicine for nourishing the kidney, liver, and spleen. It is mainly used clinically to treat kidney yin and yang deficiency syndromes such as kidney deficiency and low back pain, impotence and spermatorrhea, frequent urination, uterine coldness and infertility, dark eyes and loose stools.

[0007] Cyathula officinalis Kuan is a perennial herbaceous plant of the genus Amaranthaceae, with a neutral nature, sweet taste and slightly bitter taste. It has the functions of removing blood stasis, promoting menstruation, promoting joint function, promoting diuresis and relieving stranguria; it is mainly used to treat blood stasis, amenorrhea, abdominal masses, joint pain, traumatic injuries, hematuria, and postpartum retention of placenta. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide the application of a traditional Chinese medicine composition in the preparation of drugs for infertility and sterility.

[0009] To achieve the above object, the present invention provides the following technical solution: A traditional Chinese medicine composition, characterized in that it is composed of 30-70% of Cuscuta chinensis, 10-50% of Lycium barbarum, 10-20% of Rehmannia glutinosa, and 1-10% of Cyathula officinalis. This traditional Chinese medicine composition is preferably composed of 30-55% of Cuscuta chinensis, 17-38% of Lycium barbarum, 13-19% of Rehmannia glutinosa, and 3-6% of Cyathula officinalis.

[0010] The present invention relates to the application of a traditional Chinese medicine composition in infertility and sterility diseases, which can significantly increase the ovarian coefficient of infertile mice and significantly reduce the rate of oocyte malformation; it can promote early hatching of larvae and promote embryonic development.

[0011] In the present invention, the traditional Chinese medicine composition has undergone safety toxicological tests, 28-day oral toxicity tests, Ames tests, micronucleus tests of polychromatic erythrocytes in mouse bone marrow, and mouse sperm malformation tests, all of which show that the traditional Chinese medicine composition has no toxic effects and is safe.

[0012] Therefore, the composition of the present invention can be applied in the preparation of drugs for infertility and sterility. The present invention provides a new clinical use for this traditional Chinese medicine extract and expands its application scope. Brief Description of the Drawings

[0013] By reading the detailed description of the non-limiting embodiments with reference to the following drawings, other features, objectives, and advantages of the present invention will become more apparent

[0014] Figure 1 Determining the effect of the traditional Chinese medicine composition on embryonic development using the zebrafish model (A. model group; B. low-dose group; C. medium-dose group; D. high-dose group) Detailed Embodiments

[0015] The following will further elaborate on the present invention in combination with specific embodiments. These examples are only for illustrative purposes and do not limit the scope of the present invention.

[0016] [Example 1]

[0017] Take 120 parts of Cuscuta chinensis, 60 parts of Lycium barbarum, 35 parts of Rehmannia glutinosa, and 10 parts of Cyathula officinalis, and respectively crush them with a pulverizer to obtain their powder, with the mesh number controlled at 20 meshes. Wash the above-mentioned powdered Cuscuta chinensis, Lycium barbarum, Rehmannia glutinosa, and Cyathula officinalis with water to remove impurities; prepare the medicinal materials into 100 volumes and soak them in backflow water at 25°C for 60 minutes; extract them with a 70% ethanol solution for 60 minutes; (4) extract three times, filter and concentrate the decoction; add honey or potassium sorbate; bottle and sterilize to obtain an oral liquid based on the composition.

[0018] [Example Two]

[0019] Take 120 parts of Cuscuta chinensis, 60 parts of Lycium barbarum, 35 parts of Rehmannia glutinosa and 10 parts of Cyathula officinalis. Crush them respectively with a pulverizer to obtain their powders, and control the mesh number at 20 meshes. Wash the above-mentioned powdered Cuscuta chinensis, Lycium barbarum, Rehmannia glutinosa and Cyathula officinalis with water to remove impurities; prepare the medicinal materials into 100 volume parts, and soak them in reflux water at 25 °C for 60 minutes; perform extraction with a 70% ethanol solution for 60 minutes; extract three times, filter and concentrate the decoction; add starch and perform spray drying; granulate, package and sterilize to obtain the granule based on the composition.

[0020] [Example Three]

[0021] Take 120 parts of Cuscuta chinensis, 60 parts of Lycium barbarum, 35 parts of Rehmannia glutinosa and 10 parts of Cyathula officinalis. Crush them respectively with a pulverizer to obtain their powders, and control the mesh number at 20 meshes. Wash the above-mentioned powdered Cuscuta chinensis, Lycium barbarum, Rehmannia glutinosa and Cyathula officinalis with water to remove impurities; prepare the medicinal materials into 100 volume parts, and soak them in reflux water at 25 °C for 60 minutes; perform extraction with a 70% ethanol solution for 60 minutes; extract three times, filter and concentrate the decoction; add starch and perform spray drying; make granules, fill capsules, package and sterilize to obtain the capsule based on the composition.

[0022] [Example Four]

[0023] Take 120 parts of Cuscuta chinensis, 60 parts of Lycium barbarum, 35 parts of Rehmannia glutinosa and 10 parts of Cyathula officinalis. Crush them respectively with a pulverizer to obtain their powders, and control the mesh number at 20 meshes. Wash the above-mentioned powdered Cuscuta chinensis, Lycium barbarum, Rehmannia glutinosa and Cyathula officinalis with water to remove impurities; prepare the medicinal materials into 100 volume parts, and soak them in reflux water at 25 °C for 60 minutes; perform extraction with a 70% ethanol solution for 60 minutes; extract three times, filter and concentrate the decoction; add starch and perform spray drying; make granules, add a suitable disintegrant and lubricant, compress into tablets, package and sterilize to obtain the tablet based on the composition.

[0024] [Example Five]

[0025] Take 150 kg of Cuscuta chinensis, 180 kg of Lycium barbarum, 90 kg of Rehmannia glutinosa, and 16 kg of Cyathula officinalis. Crush them separately with a pulverizer to obtain their powdery substances, and control the mesh number at 20 meshes. Mix the above-mentioned powdered Cuscuta chinensis, Lycium barbarum, Rehmannia glutinosa, and Cyathula officinalis with 5000 kg of ethanol solution with a concentration of 75% and a pH value of 6.3, and stir evenly in a stirrer. Then, reflux by heating for 12 hours to fully dissolve the active ingredients in the original cells into the ethanol solution. Filter out the residue to obtain approximately 5000 kg of the crude solution of the Chinese patent medicine of the present invention. Place the above-mentioned crude solution in a vacuum dryer and perform low-temperature vacuum drying at -20°C. After drying for 48 hours, take it out to obtain approximately 58 kg of the dry extract of the Chinese patent medicine of the present invention. Finally, process the dry extract through pharmaceutical processing to obtain Chinese patent medicines in the forms of tablets, capsules, granules, infusion preparations, or powders.

[0026] [Example Six] Safety Toxicology Test of the Traditional Chinese Medicine Composition (Prepared by the Method of Example One)

[0027] The LD of the traditional Chinese medicine composition prepared by the present invention (prepared by the method of Example One) in the acute oral toxicity test of male and female rats 50 is greater than 10000 mg / kg·bw. According to the acute toxicity dose classification standard, this sample belongs to the actual non-toxic level; the test results of "GB 15193.22-2014 National Food Safety Standard 28-Day Oral Toxicity Test" show that: the traditional Chinese medicine composition of the present invention is administered to rats at a dose of 2 g / kg·bw / d, 4 g / kg·bw / d, and 8 g / kg·bw / d for 28 days. The general conditions of the test animals are good, and there are no abnormal changes in body weight, food utilization rate, organ weight, and organ coefficient; no abnormal changes are found in the eye examination; the results of urine indexes, hematological indexes, and biochemical indexes show that all indexes are within the normal range; no pathological changes related to the test sample are found in the pathological histological examinations of each organ. According to the 28-day oral toxicity test of GB 15193.22-2014 National Food Safety Standard, the no-observed-adverse-effect level (NOAEL) of the traditional Chinese medicine composition of the present invention is greater than 8 g / kg·bw / d (equivalent to 100 times the human recommended dose); the test results of "GB 15193.4-2014 National Food Safety Standard Bacterial Reverse Mutation Test", "GB 15193.5-2014 National Food Safety Standard Mammalian Erythrocyte Micronucleus Test", and "GB 15193.23-2014 National Food Safety Standard In Vitro Mammalian Cell Chromosome Aberration Test" show that: the Ames test, mouse bone marrow polychromatic erythrocyte micronucleus test, and mouse sperm malformation test of the traditional Chinese medicine composition of the present invention are all negative.

[0028] [Example Seven] Study on the Effect of the Traditional Chinese Medicine Composition (Prepared by the Method of Example One) on Female Infertile Mice

[0029] Busulfan is a chemotherapeutic drug that can cause mice to lose their reproductive ability and has been widely used to establish infertility models.

[0030] The experimental animals were healthy, mature Kunming white mice with normal fertility. They were fed with freely available drinking water. After a 7-day adaptation period, they were randomly divided into two groups. In the experimental group, 45 mice were intraperitoneally injected with busulfan at a concentration of 30 mg / kg (containing 3.75 ml / kg of DMSO in the solvent) once. In the control group, 15 mice were intraperitoneally injected with 3.75 ml / kg of DMSO once. After 30 days of regular feeding, using ovarian coefficient, total number of superovulated eggs / oocyte malformation rate, etc. as evaluation indicators, the effects of two doses (high and low doses, 700 and 175 mg / kg respectively) of the traditional Chinese medicine composition of the present invention on the reproductive ability of infertile mice were studied.

[0031] Ovarian coefficient = (ovary weight / body weight) * 1000;

[0032] Oocyte malformation rate = (total number of ovulated eggs - normal MII-stage cells) / total number of ovulated eggs

[0033] Table 5 Effects of the traditional Chinese medicine composition of the present invention on reproductive parameters of female mice injected with busulfan

[0034]

[0035] *P < 0.05, **P < 0.01 vs model group. △ P < 0.05, △△ P < 0.01 vs normal control group

[0036] From the reproductive data, it can be seen that when the traditional Chinese medicine extract of the present invention is at two doses (high and low), compared with the model group, it has a higher ovarian coefficient (P < 0.01) and a lower malformation rate (P < 0.01), which is comparable to the normal control group. This also means that the traditional Chinese medicine composition (prepared according to the method of Example 1) can restore the infertility state caused by busulfan to make it comparable to the normal state.

[0037] [Example 8] Determining the effect of the traditional Chinese medicine composition on embryonic development using a zebrafish model

[0038] Zebrafish were all raised in fish-raising water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450 - 550 μS / cm; the pH was 6.5 - 8.5; the hardness was 50 - 100 mg / L CaC03), and they were provided by the fish-raising center of Nanjing Huante Intelligent Fish Biotechnology Co., Ltd.

[0039] The transgenic zebrafish Tg(vas:EGFP) was reproduced by natural paired mating. At 1 dpf, fluorescent zebrafish were randomly selected and grouped, specifically: control group (added with embryo culture medium), low-concentration traditional Chinese medicine group, medium-concentration traditional Chinese medicine group, high-concentration traditional Chinese medicine group, and about 20 embryos were soaked in each group. After soaking for one day (2 dpf), about 10 zebrafish were randomly selected, fixed with low-melting-point glue and photographed.

[0040] After the transgenic zebrafish Tg(vas:EGFP) was treated with traditional Chinese medicine at different concentrations, Figure 1 it was found that the hatching rate of zebrafish larvae at 2 dpf increased significantly, indicating that the inventive composition can promote early hatching of larvae and embryonic development.

[0041] (The method refers to "Prostaglandin E2 promotes embryonic vascular development and maturation in zebrafish" Ugwuagbo KC Biology Open (2019) 8, bio039768.doi: 10.1242 / bio.039768).)

Claims

1. A traditional Chinese medicine composition applied to the preparation of infertility drugs, characterized in that it consists of 30-70% of Cuscuta chinensis, 10-50% of Lycium barbarum, 10-20% of Rehmannia glutinosa, and 1-10% of Cyathula officinalis.

2. The traditional Chinese medicine composition according to claim 1, characterized in that the pharmaceutically acceptable carrier includes one or a combination of sucrose, dextrin, starch, lactose, mannitol, xylitol, chitosan, bifidose, soluble starch, talc powder, water-soluble dextrin, sodium carboxymethylcellulose (CMS-Na), microcrystalline cellulose, colloidal silica, hydroxypropyl starch, ethanol, hydroxypropyl cellulose, polyethylene glycol, chitosan, polyvinylpyrrolidone.

3. The traditional Chinese medicine composition according to claim 1, characterized in that preferably it consists of 30-55% of Cuscuta chinensis, 17-38% of Lycium barbarum, 13-19% of Rehmannia glutinosa, and 3-6% of Cyathula officinalis.

4. The application according to claim 1, characterized in that the traditional Chinese medicine composition can significantly increase the ovarian coefficient of infertility and can significantly reduce the malformation rate of oocytes.

5. The application according to claim 1, characterized in that the traditional Chinese medicine composition can promote the early development of larvae.

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