Traditional Chinese medicine composition for male oligospermia and preparation method and application thereof

Abstract

The invention relates to a traditional Chinese medicine composition for male oligospermia as well as a preparation method and application thereof, and relates to the technical field of traditional Chinese medicines. The traditional Chinese medicine composition comprises ginseng, cornu cervi pantotrichum, codonopsis pilosula, angelica sinensis, prepared rehmannia root, astragalus membranaceus, radix paeoniae alba, ligusticum wallichii, corydalis tuber, fenugreek, teasel root, bighead atractylodes rhizome, rhizoma cyperi, fructus amomi, leonurus, scutellaria baicalensis, parasitic loranthus and honey-fried licorice root. The traditional Chinese medicine composition can treat male oligospermia well by improving testis injury, raising sperm motility and adjusting sex hormone balance.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for male oligospermia and a preparation method and application thereof. BACKGROUND

[0003] General treatment of oligospermia: weight loss, smoking cessation, alcohol cessation, physical exercise can improve abnormal sperm parameters, hormone levels and obesity-related secondary hypogonadism, avoid long-term environmental exposure factors and stop using drugs affecting spermatogenesis function. Modern medicine has many treatment methods for male infertility, such as hormones and antioxidants. Antioxidants play an important role in the treatment of male infertility, but they have many side effects and the problem of poor treatment effect when used alone. For example, excessive use of antioxidants can cause oxidative and reduction system dysfunction, thereby causing damage to the human body. This process is known as the "antioxidant paradox". The use of hormones also has the problem of poor efficacy, and long-term use can cause gastrointestinal irritation, drug-induced obesity and other adverse reactions. Moreover, hormone therapy is not suitable for all patients. SUMMARY

[0004] In view of the above problems, the present application provides a traditional Chinese medicine composition for male oligospermia, which can better treat male oligospermia by improving testicular damage, restoring sperm motility and regulating sex hormone balance.

[0005] In order to achieve the above purpose, the present application provides a traditional Chinese medicine composition for male oligospermia, and the effective components of the traditional Chinese medicine composition are mainly prepared from the following raw materials in a weight ratio:

[0006]

[0007]

[0008] The present inventors found in the research that after excluding organic lesions such as genetic diseases, chromosomal abnormalities, varicocele and the like, traditional Chinese medicine has a unique treatment strategy and good curative effect in treating oligospermia and asthenospermia. Traditional Chinese medicine believes that the kidney is the root of preheaven, and is responsible for storing essence and reproductive development, so kidney tonification and essence tonification are the basic treatment methods for treating such patients according to syndrome differentiation. Kidney blood stasis, blood essence blocking collaterals and blood stasis in the essence chamber are the main causes of male semen non-formation, sperm reduction, disappearance, deformity and infertility, and the use of kidney tonifying and essence tonifying, qi and blood activating drugs to achieve the effects of kidney tonification, essence and blood tonification and qi tonification can improve male oligospermia / azoospermia and other infertility symptoms. At the same time, modern pharmacological research believes that male oligospermia is closely related to the ability of testicular tissue to produce sperm. Sperm reduction or disappearance caused by abnormal testicular tissue is the main cause of male fertility reduction. Serum sex hormones such as luteinizing hormone (LH), testosterone (T) and follicle-stimulating hormone (FSH) are important factors affecting testicular spermatogenic function. Testis is an important male reproductive organ, and its histopathological changes and quality reduction will not only cause sperm reduction and sperm deformity, but also affect the male spermatogenic function and fertility.

[0009] Therefore, the present inventors propose the above-mentioned traditional Chinese medicine composition - Shenshuangbaifeng pills, which is composed of ginseng, deer horn, radix codonopsis, angelica sinensis, prepared rehmannia, astragalus root, white peony root, chuanxiong, rhizoma corydalis, fenugreek, radix dipsaci, white atractylodes, cyperus rotundus, motherwort, radix scrophulariae, taxillus, and licorice, has the effects of tonifying qi and blood and regulating menstruation, is used for treating deficiency of qi and blood, irregular menstruation and menstrual abdominal pain, and has been a holy medicine for gynecology. However, the traditional Chinese medicine composition meets the requirements of traditional Chinese medicine theory, method and prescription for treating male oligospermia, and ginseng is the monarch drug for tonifying primordial qi, strengthening the spleen and lung, deer horn is the monarch drug for tonifying kidney yang, tonifying essence and blood, strengthening bones and muscles, regulating chong and ren, astragalus root, radix codonopsis and white atractylodes have the effects of tonifying qi and invigorating the spleen and stomach; angelica sinensis, white peony root, chuanxiong, prepared rehmannia have the effects of tonifying blood and activating blood; fenugreek, taxillus and radix dipsaci have the effects of tonifying liver and kidney and strengthening bones and muscles, which are the ministerial drugs; cyperus rotundus, radix scrophulariae and radix scrophulariae have the effects of regulating qi and activating blood, which are the assistant drugs; and licorice regulates all drugs and makes the drugs work together. The present application mainly tonifies yang and kidney, tonifies qi and blood, regulates menstruation, relieves pain, clears heat, removes dampness and tonifies kidney, so that the present application tonifies without stagnation, warms without dryness, achieves the good effects of tonifying qi and blood, regulating qi and activating blood and tonifying kidney essence.

[0010] At present, the existing technology has disclosed the traditional Chinese medicine composition for treating oligospermia and asthenospermia, which is composed of ginseng, deer antler, astragalus, turtle shell glue, prepared rehmannia, radix lactiferae, Chinese yam, salvia miltiorrhiza, medlar, cuscuta, schisandra and taxillus. There is also a composition of ginseng, astragalus, angelica, Chinese yam, deer antler glue, medlar, bupleurum, cimicifuga, and licorice. The commercially available Chinese medicine Shengjing capsule is composed of deer antler, medlar, ginseng, cordyceps, cuscuta, sandarac, epimedium, polygonatum, radix polygoni multiflori, mulberry, psoralea, dracaena, herba, euonymus, aronia, eucommia, radix morindae officinalis, lonicera japonica, and ginkgo biloba. It has the effects of tonifying kidney and essence, nourishing yin and strengthening yang. The above-mentioned patent compositions are basically tonic medicine, while the above-mentioned composition of the present application has the effects of warming kidney and dispelling cold, regulating qi and promoting blood circulation, clearing heat and removing dampness, and tonifying yang and kidney, benefiting qi and blood, and stopping pain, clearing heat, removing dampness, and tonifying kidney. The traditional Chinese medicine composition of the present application is tonifying but not stagnating, warming but not drying, and has the good effects of benefiting qi and blood, regulating qi and promoting blood circulation, and tonifying kidney essence. It also treats oligospermia and asthenospermia caused by damp-heat, phlegm turbidity, and blood stasis. In addition, the query of the traditional Chinese medicine composition for oligospermia mostly involves medlar, Chinese yam, poria, polygonatum, plantago, schisandra, and cuscuta. These traditional Chinese medicines are mostly food and medicine homologous traditional Chinese medicines, which are used for diet therapy and mainly for conditioning. The treatment effect is not clear, and the drug efficacy is not enough. The traditional Chinese medicine composition of the present application takes ginseng and deer antler as the monarch drug. Ginseng is the first product for benefiting qi, and deer antler is the first drug for tonifying kidney. At the same time, angelica, radix paeoniae alba, chuanxiong, and prepared rehmannia are the classic four-ingredient decoction, which is the first formula for tonifying blood. Dipsacus and taxillus are a classic drug pair for tonifying kidney. Dipsacus tonifies liver and kidney and regulates blood vessels, and has the effect of tonifying but not stagnating, and mainly warms and tonifies kidney yang. Taxillus benefits blood vessels and mainly nourishes yin blood. The combination of the two drugs tonifies liver and kidney, and balances yin and yang, so that the effect of tonifying liver and kidney is more obvious. The whole prescription has the effects of benefiting qi and blood, tonifying kidney and filling essence, and can greatly improve the male spermatogenesis function and fertility. Other studies use turtle shell glue to treat oligospermia or use radix polygoni multiflori to tonify liver and kidney to treat oligospermia. However, the turtle shell glue on the market is difficult to distinguish between true and false, and the drug efficacy is difficult to control. The hepatotoxicity of radix polygoni multiflori is a concern. In contrast, the traditional Chinese medicine composition of the present application has no toxicity, small side effects, and high safety. Therefore, the traditional Chinese medicine composition of the present application has unique properties, and the expected effect is good.

[0011] Moreover, from the perspective of modern pharmacology, the traditional Chinese medicine composition of the present application also has the effects of enhancing sexual gland function, improving reproductive ability, and improving testicular damage.

[0012] Ginseng has the pharmacological effect of enhancing the function of sex glands: Ginseng has the effect of stimulating the function of hypothalamus-pituitary-gonadal axis. Ginseng and ginsenosides have the effect of stimulating the secretion of gonadotropin from pituitary, accelerating the sexual maturation of rats, accelerating the appearance of estrus in immature female mice, increasing the weight of uterus and ovary, and increasing the secretion of luteinizing hormone. Ginseng also increases the weight of testis and epididymis in young male animals, expands the diameter of vas deferens, increases the number of sperm in testis, increases the motility of sperm, prolongs the survival period of sperm in vitro, and causes copulation in castrated rats.

[0013] Pilose antler has the effect of sex hormone: Pilose antler has the effect of male hormone and female hormone. Pilose antler contains estradiol which has the activity of female hormone, can promote the development of reproductive system in female young mice, increase the weight of uterus and ovary, and cause compensatory hyperplasia and changes in uterus and vagina of castrated rats. Subcutaneous injection of pilose antler extract in immature female mice can promote the development of ovary and increase the weight of ovary. Ethanol extract of pilose antler can increase the number of spermatogonia and the number of spermatogenic cell layers in rats. D component extracted from pilose antler can increase the weight of testis, epididymis, prepuce gland, prostate and semen in mice with yang deficiency and bone marrow damage. Pilose antler extract can significantly increase the weight of testis, prostate and seminal vesicle in immature male animals (rats and mice). Gastric administration of ethanol extract of pilose antler to aging mice can significantly increase the content of testosterone in plasma of aging mice, but has no obvious effect on the content of testosterone in plasma of normal mice. Pilose antler extract injection can promote the growth of prostate, seminal vesicle and prepuce gland in castrated rats.

[0014] Codonopsis pilosula has a certain effect on the reproductive capacity of rats, and the polysaccharide in Codonopsis pilosula can promote the secretion of testosterone hormone in rat testicular interstitial cells in vitro. At the same time, it can significantly reduce the activity of SOD, reduce the content of MDA, and enhance the activity of GSH-Px in rat interstitial cells in vitro, promote sperm maturation, and increase sperm count.

[0015] Radix astragali has the pharmacological effect of enhancing the function of sex glands: Radix astragali can prolong the estrus period of mice and has good effect on the development of mice. Radix astragali can enhance sperm motility. The addition of Radix astragali decoction in human semen in vitro can significantly improve sperm motility, sperm movement speed, sperm forward movement speed, and frequency of sperm head swing.

[0016] The main component of Radix Dipsaci, Radix Dipsaci saponin, can improve sperm cell apoptosis, increase sperm count, and improve sperm motility.

[0017] Horse gram contains trigonelline which can significantly improve the damage degree of testis of mice caused by heat stress.

[0018] Therefore, we construct the mouse model of oligospermia induced by cyclophosphamide, and detect the weight of mice, testis index, sperm motility, pathological state of testis and epididymis, and change of hormone level related to reproductive function. The animal experiment proves that the above-mentioned traditional Chinese medicine composition can increase the content level of testosterone, luteinizing hormone and follicle stimulating hormone in serum, thereby repairing the damage of seminiferous tubules, improving the damage of testis, increasing the spermatogenic cells, increasing the number of sperm in epididymis, rising the sperm motility, regulating the balance of sex hormones, and further treating male oligospermia.

[0019] In one of the embodiments, the effective components of the traditional Chinese medicine composition are prepared from the following raw materials by weight ratio:

[0020]

[0021] In one of the embodiments, the Radix Astragali is Radix Astragali (prepared with wine), the Radix Paeoniae Alba is Radix Paeoniae Alba (prepared with wine), the Ligusticum Chuanxiong is Ligusticum Chuanxiong (prepared with wine), the Rhizoma Corydalis is Rhizoma Corydalis (prepared), the Trigonella foenum-graecum is Trigonella foenum-graecum (prepared with salt), the Radix Dipsaci is Radix Dipsaci (prepared with wine), the Rhizoma Atractylodis Macrocephalae is Rhizoma Atractylodis Macrocephalae (prepared), the Herba Leonuri is Herba Leonuri (prepared with wine), the Radix Scrophulariae is Radix Scrophulariae (prepared with wine), the Morinda officinalis is Morinda officinalis (steamed), the Cornu Cervi is Cornu Cervi (prepared with wine), the Radix Codonopsis is Radix Codonopsis (prepared with wine), the Angelica sinensis is Angelica sinensis (prepared with wine), and the Cyperus rotundus is Cyperus rotundus (prepared).

[0022] The application further provides a preparation method of the traditional Chinese medicine composition, which comprises the following steps: mixing Radix Rehmanniae Preparata, Radix Astragali, Radix Paeoniae Alba, Ligusticum Chuanxiong, Rhizoma Corydalis, Trigonella foenum-graecum, Radix Dipsaci, Rhizoma Atractylodis Macrocephalae, Herba Leonuri, Radix Scrophulariae, Morinda officinalis, and Radix Glycyrrhizae, adding water, decocting, taking decocted liquid, filtering, concentrating, obtaining thick paste, crushing Cornu Cervi, Radix Ginseng, Radix Codonopsis, Angelica sinensis, Cyperus rotundus, and Amomum villosum, and mixing the crushed materials with the thick paste to obtain the traditional Chinese medicine composition.

[0023] In one of the embodiments, the preparation method comprises the following steps: mixing Radix Rehmanniae Preparata, Radix Astragali (prepared with wine), Radix Paeoniae Alba (prepared with wine), Ligusticum Chuanxiong (prepared with wine), Rhizoma Corydalis (prepared), Trigonella foenum-graecum (prepared with salt), Radix Dipsaci (prepared with wine), Rhizoma Atractylodis Macrocephalae (prepared), Herba Leonuri (prepared with wine), Radix Scrophulariae (prepared with wine), Morinda officinalis (steamed), and Radix Glycyrrhizae, adding 10 times of water, decocting, combining decocted liquid, filtering, reducing pressure and concentrating, heating and concentrating to obtain thick paste; crushing Cornu Cervi (prepared with wine) to obtain Cornu Cervi (prepared with wine) fine powder, crushing Radix Ginseng, Radix Codonopsis (prepared with wine), Angelica sinensis (prepared with wine), Cyperus rotundus (prepared), and Amomum villosum in series, uniformly mixing the crushed materials with the Cornu Cervi (prepared with wine) fine powder, and mixing the crushed materials with the thick paste to obtain the traditional Chinese medicine composition.

[0024] In one of the embodiments, the decocting comprises: decocting twice, decocting for 3-5 hours in the first time, and decocting for 1-3 hours in the second time; and the relative density of the thick paste is 1.2-1.3.

[0025] The application further provides application of the traditional Chinese medicine composition in preparation of a medicine for treating male oligospermia.

[0026] In one of the embodiments, the medicine can repair the damage of the seminiferous tubule, improve the damage of the testis, increase the spermatogenic cells, increase the sperm quantity of the epididymis, restore the sperm activity, and regulate the sex hormone balance by increasing the content of testosterone, luteinizing hormone and follicle stimulating hormone in the serum, thereby treating the oligospermia of the male.

[0027] The application further provides a medicine for treating the oligospermia of the male, which comprises a pharmaceutical active ingredient and excipients, wherein the pharmaceutical active ingredient is the traditional Chinese medicine composition.

[0028] In one of the embodiments, the excipients comprise at least one of a solvent, a disintegrant, a flavoring agent, a coloring agent, a lubricant, an antioxidant, a preservative, a binder, a filler or a thickening agent.

[0029] In one of the embodiments, the dosage form of the medicine is oral liquid, pill, granule, capsule, tablet, dripping pill, injection or powder.

[0030] Compared with the prior art, the application has the following beneficial effects:

[0031] The traditional Chinese medicine composition for the oligospermia of the male can treat the oligospermia of the male by improving the damage of the testis, restoring the sperm activity and regulating the sex hormone balance. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 Testis picture of Example 2;

[0033] Figure 2 Testis index statistical result picture of Example 2;

[0034] Figure 3 Sperm activity statistical result picture of Example 2;

[0035] Figure 4 Testis HE staining result picture of Example 2;

[0036] Figure 5 Epididymis HE staining result picture of Example 2;

[0037] Figure 6 Kidney HE staining result picture of Example 2;

[0038] Figure 7 ELISA detection result picture, wherein the first picture from top to bottom is a statistical picture of the content change of serum T of the mice in each group, the second picture is a statistical picture of the content change of serum LH of the mice in each group, and the third picture is a statistical picture of the content change of serum FSH of the mice in each group. DETAILED DESCRIPTION

[0039] For the purposes of promoting an understanding of the present application, the present application will be described in greater detail below with reference to the illustrative embodiments. In order to facilitate an understanding of this application, a thorough and complete description of the salient, non- limiting aspects of the application is provided. Nevertheless, it is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting. As used in this specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the content clearly dictates otherwise. The term "and / or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.

[0041] The reagents, materials and equipment used in the examples are commercially available unless otherwise specified. The test methods are conventional test methods in the art unless otherwise specified.

[0042] Example 1

[0043] The present application provides a preparation method of a traditional Chinese medicine composition for treating oligospermia in men, comprising the following steps:

[0044] Radix Rehmanniae 80 parts, Radix Paeoniae Alba (prepared with wine) 40 parts, Radix Angelicae Sinensis (prepared with wine) 40 parts, Ligusticum Chuanxiong (prepared with wine) 35 parts, Rhizoma Corydalis (prepared) 30 parts, Trigonella foenum-graecum (prepared with salt) 35 parts, Dipsacus asper (prepared with wine) 35 parts, Rhizoma Atractylodis Macrocephalae (prepared) 40 parts, Leonurus japonicus (prepared with wine) 30 parts, Radix Paeoniae Alba (prepared with wine) 35 parts, Taxillus sutchuenensis (steamed) 30 parts, Radix Glycyrrhizae (prepared with wine) 35 parts are added with 10 times the amount of water, decocted twice, the first time for 4 hours and the second time for 2 hours, the decoction is combined, filtered, concentrated under reduced pressure, further heated and concentrated into a thick paste with a relative density of 1.2-1.3; Cornu Cervi (prepared with wine) 10 parts is separately pulverized into fine powder, Panax ginseng 10 parts, Radix Codonopsis (prepared with wine) 45 parts, Radix Angelicae Sinensis (prepared with wine) 40 parts, Rhizoma Cyperi (prepared) 35 parts, and Fructus Amomi 30 parts are stringed and pulverized, mixed with the fine powder of Cornu Cervi, and then mixed with the thick paste to obtain the traditional Chinese medicine composition.

[0045] Example 2

[0046] 1. Experimental method

[0047] 1.1 Experimental modeling and grouping

[0048] Kunming mice 48 were randomly divided into normal group, model group, traditional Chinese medicine composition low dose group, traditional Chinese medicine composition medium dose group, traditional Chinese medicine composition high dose group and positive drug Shengjing capsule group (Shengjing capsule, GMP: Z20027672, batch number: 20230410, purchased from Zunyi Liao Yuanhe Hall Pharmaceutical Co., Ltd.), 8 in each group. Except for the normal group, the rest of the mice were adaptively fed for one week before modeling, and then modeling was started. The modeling drug cyclophosphamide was dissolved in normal saline, and was given by intraperitoneal injection at a dose of 75 mg / kg, with a dose of 0.1 ml / 10 g. The modeling lasted for 7 days.

[0049] After modeling, the model group was not given drug, and among the three groups of traditional Chinese medicine compositions, the medium dose was set as the clinical equivalent dose group, and the low dose and the high dose were 1 / 2 and 2 times of the clinical equivalent dose respectively. Each treatment group started giving drug on the first day of modeling, and was given corresponding dose and drug according to the group. The high dose group was given traditional Chinese medicine composition 2.444 g·kg -1 ·d -1 , the medium dose group was given traditional Chinese medicine composition 1.222 g·kg -1 ·d -1 , the low dose group was given traditional Chinese medicine composition 0.0611 g·kg -1 ·d -1 , the positive drug group was given Shengjing capsule 0.624 g·kg -1 ·d -1 , and the drug administration lasted for 14 days.

[0050] 1.2 Body weight detection.

[0051] All mice were weighed and the body weight was recorded before modeling started. The body weight of mice in each group was recorded every day after modeling started, and lasted for 14 days.

[0052] 1.3 Specimen collection and preparation.

[0053] After the last administration, the mice were fasted for 8 h, and then the samples were taken. After the mice were weighed and recorded, the blood was collected by enucleation method under 3% sodium pentobarbital anesthesia. After standing for 4 h, the upper serum was collected after centrifugation at 3000 r / min for 10 min. The left testis was placed in testicular fixative, and the right testis was frozen. The left epididymis was placed in testicular fixative, and the right epididymis was subjected to sperm count to detect sperm motility. The bilateral kidneys were collected, and the left kidney was placed in 4% paraformaldehyde fixative, and the right kidney was frozen.

[0054] 1.4 Testis index.

[0055] After the last administration, the bilateral testes were weighed, and the testis index was calculated, i.e. testis index = bilateral testis weight (mg) / body weight (g).

[0056] 1.5 Sperm motility detection.

[0057] Take the mouse right epididymis cut into centrifuge tube, then add 37℃ water bath preheated saline 200 μL, with pipette gun gently blow mix uniform, in 37℃ water bath incubator for 5 min, then take sperm suspension placed in blood cell counter counting. First counting, blood cell counter placed in 60℃ oven heating 5 min, take out after counting total sperm number. Sperm motility = (total sperm number - dead sperm number) / total sperm number.

[0058] 1.6 HE staining.

[0059] Take Bouin's fixed liquid fixed 24 h testis tissue, carry out conventional transparency, wax immersion, embedding, section (5 μm). After dewaxing, hydration of the section, according to the HE kit instructions, conventional staining, neutral optical resin mounting under the microscope to observe the morphological changes of tissue.

[0060] 1.7 ELISA detection.

[0061] Serum T, LH, FSH is an important factor affecting testicular spermatogenic function. Serum T, LH, FSH level changes may be used as an important indicator of evaluation of testicular spermatogenic effect. After taking the mouse serum according to the ELISA instruction book operation, the OD value of each group was detected on the enzyme label instrument, and the serum FSH, LH, T content was obtained.

[0062] 2 Experimental results.

[0063] 2.1 Body weight change trend.

[0064] Table 1 body weight change trend of each group of mice (unit: g, n = 8)

[0065]

[0066] 2.2 testis index, testis picture.

[0067] Testis picture as Figure 1 shown, testis index as Figure 2 , table 2.

[0068] Table 2 testis index of each group of mice n = 8)

[0069]

[0070] Compared with the normal group, P < 0.01 marked * ; compared with the model group, P < 0.05 marked # , P < 0.01 marked ## .

[0071] 2.3 sperm motility calculation.

[0072] Results as shown in Table 3. Figure 3

[0073] Sperm motility of each group of mice in Table 3 n = 8

[0074]

[0075] Compared with the normal group, P <0.05 marked *; compared with the model group, P <0.05 marked # .

[0076] 2.4 HE staining results.

[0077] 2.4.1 Testis HE (200X).

[0078] Results as shown in Table 3. Figure 4 The testis of the normal group of mice was closely arranged, the lumen was regular, the lumen was regularly arranged, and mature sperm was seen; compared with the normal group, the testis of the model group of mice was irregular, the lumen of the testis was irregular, and the lumen of the testis was irregular. The number of spermatogenic cells in the lumen was significantly reduced, and part of the spermatogenic tubules only contained supporting cells; after treatment with the traditional Chinese medicine composition, the testicular damage of each group of mice was improved, among which the low-dose group appeared in the spermatogenic tubules, mainly in the spermatogenic tubules. Spermatogenic cells appeared in the lumen of the testis of the medium-dose group, and most of the spermatogenic cells appeared in the lumen of the testis of the high-dose group, and mature sperm was seen; the same as the positive drug treated mice testis, the testis was regularly arranged, and the gap was small. Most of the spermatogenic cells in the lumen of the testis were restored to multiple layers, regularly arranged, and mature sperm was seen.

[0079] 2.4.2 Epididymis HE (200X).

[0080] Results as shown in Table 3. Figure 5 The testis of the normal group of mice was closely arranged, the lumen was regular, the lumen was regularly arranged, and mature sperm was seen; compared with the normal group, the testis of the model group of mice was irregular, the lumen of the testis was irregular, and the lumen of the testis was irregular. The number of spermatogenic cells in the lumen was significantly reduced, and part of the spermatogenic tubules only contained supporting cells; after treatment with the traditional Chinese medicine composition, the testicular damage of each group of mice was improved, among which the low-dose group appeared in the spermatogenic tubules, mainly in the spermatogenic tubules. Spermatogenic cells appeared in the lumen of the testis of the medium-dose group, and most of the spermatogenic cells appeared in the lumen of the testis of the high-dose group, and mature sperm was seen; the same as the positive drug treated mice testis, the testis was regularly arranged, and the gap was small. Most of the spermatogenic cells in the lumen of the testis were restored to multiple layers, regularly arranged, and mature sperm was seen.

[0081] 2.4.3 Kidney HE (200X).

[0082] Results as shown in Table 3. Figure 6 ​As shown, the glomerular structure of normal group mice was normal, and the glomerular capsule space was clear; compared with the normal group, the glomerulus of the model group mice was enlarged, the mesangial cells proliferated, part of the cells were vacuolated, a small amount of inflammatory cells infiltrated, the glomerular capsule space was not clear, and red staining material was deposited in the capsule; the periglomerular interstitium had mild fibrous tissue proliferation; part of the renal tubular epithelial cells were granular and vacuolar degeneration; after treatment with the traditional Chinese medicine composition and the positive drug, compared with the model group, the above several aspects of each group of mice were improved, and the high-dose group improved most obviously.

[0083] 2.5 ELISA detection results.

[0084] Results are shown in Table 4. Figure 7

[0085] Table 4 Change of serum T, LH, FSH content of mice in each group n = 8

[0086]

[0087] Compared with the normal group, P < 0.05 is marked as *, P < 0.01 is marked as * ; compared with the model group, P < 0.05 is marked as # , P <.01 is marked as ## .

[0088] 3. Summary.

[0089] From the body weight, the initial body weight of mice in each group was in the range of 26-27 g. During the experiment, the body weight of mice in the blank group showed an upward trend, the body weight of mice in the model group and the low-dose traditional Chinese medicine composition group showed a downward trend first and then an upward trend, the body weight of mice in the medium-dose traditional Chinese medicine composition group, the high-dose traditional Chinese medicine composition group and the Shengjing capsule group showed a gentle upward trend. By comparing the body weight of mice in each group on the 1st day, the 7th day and the 14th day of administration, we observed that the body weight of mice in the model group decreased significantly compared with the blank group, and the body weight of mice in each administration group was well maintained, but the body weight change between each group was not significant. The body weight detection results showed that the traditional Chinese medicine composition could alleviate the body weight loss caused by cyclophosphamide to a certain extent, and had a potential role in protecting the function of the body, among which the medium-dose group performed best.

[0090] ​The testis index and sperm motility results show that the reproductive function damage induced by cyclophosphamide can cause the testis weight to decrease (P<0.01), the number of dead sperm to increase and the sperm motility function to be damaged (P<0.05), and after taking the traditional Chinese medicine composition, the testis index and sperm motility rebound (P<0.01 or P<0.05), the traditional Chinese medicine composition presents a dose-effect relationship, and the high-dose group of the traditional Chinese medicine composition has the best improvement effect, which shows that the traditional Chinese medicine composition has good effects in improving testis damage and restoring reproductive function. The pathological staining results show that compared with the blank group, the model group of mice has obvious damage to the testis, the number of spermatogenic cells is obviously reduced, the shape of the epididymal tube lumen is changed, and the number of sperm in the lumen is reduced, which shows that the testis and epididymis are obviously damaged. After treatment with the traditional Chinese medicine composition, the testis damage of the mice in each group is improved, among which, the low-dose group has spermatogonia in part of the seminiferous tubule lumen, the medium-dose group and the high-dose group have spermatogenic cells of all levels in the seminiferous tubule lumen, and in addition, the high-dose group also has mature sperm; and in the epididymis, the number of sperm in the lumen of the mice in each treatment group of the traditional Chinese medicine composition increases, among which, the high-dose group is the most obvious. In summary, the traditional Chinese medicine composition group can better improve the pathological state of testis damage.

[0091] The changes of the contents of testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the serum of mice can reflect the function state of the hypothalamus-pituitary-gonadal axis (HPG axis), and then reveal the pathological changes related to reproductive function damage. Among them, T is a male hormone secreted by related cells in the testis, which is responsible for maintaining male reproductive function (such as spermatogenesis) and secondary sexual characteristics; LH and FSH are usually secreted by the anterior pituitary, the former can stimulate the related cells in the testis of male mice to secrete testosterone, and the latter can act on the Sertoli cells of male mice to promote spermatogenesis. Compared with the blank group, the serum contents of T, LH and FSH of the model group of mice all decrease obviously (P<0.05, P<0.01), which shows that the reproductive function of mice is obviously damaged. After taking the traditional Chinese medicine composition, the serum contents of T, LH and FSH all rebound obviously, which presents a dose-effect relationship, among which, the high-dose group of T and LH indicators has the most obvious rebound (P<0.05, P<0.01), and the high-dose group of FSH indicators has the best improvement effect, although the result is not significant.

[0092] Therefore, the traditional Chinese medicine composition in the present application has certain improvement effects on the decrease of body weight, testis damage, decrease of sperm motility and decrease of male hormone related indicators of the azoospermia model mice caused by cyclophosphamide.

[0093] The technical features of the above-described embodiments can be combined in any manner, and to make the description concise, all possible combinations of the technical features in the above-described embodiments are not described, however, as long as the combinations of the technical features do not exist contradictions, they should be considered as the scope of the present disclosure.

[0094] The above embodiments only express several implementation manners of the present application, which are described in a more specific and detailed manner, but cannot be understood as a limitation on the patent scope of the present application. It should be noted that, for ordinary skilled persons in the art, several modifications and improvements can be made without departing from the concept of the present application, which all belong to the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. A traditional Chinese medicine composition for oligospermia in men, characterized in that, The active ingredients of this traditional Chinese medicine composition are mainly prepared from the following raw materials in the following weight ratios:

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The active ingredients of this traditional Chinese medicine composition are mainly prepared from the following raw materials in the following weight ratios:

3. The traditional Chinese medicine composition according to any one of claims 1-2, characterized in that, The Astragalus membranaceus is processed with wine, the Paeonia lactiflora is processed with wine, the Ligusticum chuanxiong is processed with wine, the Corydalis yanhusuo is processed with wine, the Trigonella foenum-graecum is processed with salt, the Dipsacus asper is processed with wine, the Atractylodes macrocephala is processed with wine, the Leonurus japonicus is processed with wine, the Scutellaria baicalensis is processed with wine, the Taxillus chinensis is steamed, the deer antler is processed with wine, the Codonopsis pilosula is processed with wine, the Angelica sinensis is processed with wine, and the Cyperus rotundus is processed with wine.

4. The method for preparing the traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, Includes the following steps: Prepared Rehmannia glutinosa, Astragalus membranaceus, Paeonia lactiflora, Ligusticum chuanxiong, Corydalis yanhusuo, Trigonella foenum-graecum, Dipsacus asper, Atractylodes macrocephala, Leonurus japonicus, Scutellaria baicalensis, Taxillus chinensis, and Glycyrrhiza uralensis are mixed, water is added, and the decoction is taken, filtered, and concentrated to obtain a thick paste. Deer antler, ginseng, Codonopsis pilosula, Angelica sinensis, Cyperus rotundus, and Amomum villosum are pulverized and mixed with the thick paste to obtain a traditional Chinese medicine composition.

5. The preparation method according to claim 4, characterized in that, The preparation method includes the following steps: Mix Rehmannia glutinosa (processed with wine), Astragalus membranaceus (processed with wine), Paeonia lactiflora (processed with wine), Ligusticum chuanxiong (processed with wine), Corydalis yanhusuo (processed), Trigonella foenum-graecum (processed with salt), Dipsacus asper (processed with wine), Atractylodes macrocephala (processed with wine), Leonurus japonicus (processed with wine), Scutellaria baicalensis (processed with wine), Taxillus chinensis (steamed), and Glycyrrhiza uralensis (processed with wine), add 10 times the amount of water, decoct, combine the decoctions, filter, concentrate under reduced pressure, and concentrate by heating to obtain a thick paste; pulverize deer antler (processed with wine) to obtain fine deer antler (processed with wine) powder; pulverize ginseng, Codonopsis pilosula (processed with wine), Angelica sinensis (processed with wine), Cyperus rotundus (processed), and Amomum villosum, mix with the fine deer antler (processed with wine) powder, and mix with the thick paste to obtain the traditional Chinese medicine composition.

6. The use of the traditional Chinese medicine composition according to any one of claims 1-3 in the preparation of a medicament for treating male oligospermia.

7. Use according to claim 6, characterized in that, The drug treats male oligospermia by increasing the levels of testosterone, luteinizing hormone, and follicle-stimulating hormone in the serum, thereby repairing seminiferous tubule damage, improving testicular damage, increasing spermatogenic cells, increasing sperm count in the epididymis, restoring sperm motility, and regulating sex hormone balance.

8. A medicament for treating oligozoospermia in men, characterized by, It includes a pharmaceutical active ingredient and excipients, wherein the pharmaceutical active ingredient is a traditional Chinese medicine composition according to any one of claims 1-3.

9. The medicament according to claim 8, characterized in that, The excipients include at least one of solvents, disintegrants, flavoring agents, colorants, lubricants, antioxidants, preservatives, binders, fillers, or thickeners.

10. The medicament according to claim 8, characterized in that, The dosage form of the drug is oral liquid, pill, granule, capsule, tablet, drop pill, injection or powder.

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