CN103239624B - Chinese medicine for treating oligospermia and asthenozoospermia owning to insufficiency of both spleen and kidney - Google Patents
Chinese medicine for treating oligospermia and asthenozoospermia owning to insufficiency of both spleen and kidney Download PDFInfo
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Abstract
The invention relates to the technical field of traditional Chinese medicines and in particular relates to a Chinese medicine for treating male infertility as well as a preparation method and an application thereof. The Chinese medicine composition is prepared from the following Chinese medicine raw materials in weight proportion: 30-600g of milkvetch, 10-300g of radix ophiopogonis, 10-300g of morinda officinalis, 5-120g of herba epimedii, 20-600g of prepared polygonum multiflorum, 30-600g of semen cuscutae and 20-500g of prepared sealwort. The Chinese medicine composition provided by the invention can be used for treating oligospermia and asthenozoospermia owning to insufficiency of both spleen and kidney.
Description
Technical field
The present invention relates to technical field of Chinese medicines, particularly relate to a kind of Chinese medicine composition for the treatment of male infertility and its preparation method and application.
Background technology
Male infertility is common clinically disease in the male sexual system, is subject to the impact of the many factors such as environment.In infertile couples, male factor accounts for half ratio, wherein take again few azoospermia as common type.In recent years, about being more and more concerned with Chinese medicine preparation treatment male research few, azoospermia.The relative Western medicine of kidney-nourishing tcm drug that the traditional Chinese medical science is used is safer.
Summary of the invention
Object of the present invention aims to provide a kind of Chinese medicine composition for the treatment of oligospermia and its preparation method and application.
Specifically, the invention provides a kind of Chinese medicine composition for the treatment of oligospermia, it is characterized in that, it is to make with the raw material of Chinese medicine of following weight proportion:
The Radix Astragali 30～600g, Radix Ophiopogonis 10～300g, Radix Morindae Officinalis 10～300g, Herba Epimedii 5～120g, Radix Polygoni Multiflori Preparata 20～600g, Semen Cuscutae 30～600g, Rhizoma Polygonati (processed) 20～500g.
In a preference, the weight proportion of described raw material of Chinese medicine is:
The Radix Astragali 60～240g, Radix Ophiopogonis 30～150g, Radix Morindae Officinalis 30～150g, Herba Epimedii 9～50g, Radix Polygoni Multiflori Preparata 50～300g, Semen Cuscutae 60～300g, Rhizoma Polygonati (processed) 50～200g.
In another preference, the weight proportion of described raw material of Chinese medicine is:
Radix Astragali 120g, Radix Ophiopogonis 60g, Radix Morindae Officinalis 60g, Herba Epimedii 20g, Radix Polygoni Multiflori Preparata 100g, Semen Cuscutae 160g, Rhizoma Polygonati (processed) 100g.
A second aspect of the present invention has been to provide the application of above-mentioned Chinese medicine composition in the medicine of preparation treatment oligospermia.
A third aspect of the present invention has been to provide the preparation method of above-mentioned Chinese medicine composition, and it comprises the following steps: to get each crude drug mixes, decocting 2～3 times, and each 1～3 hour, add 4～20 times of amounts of water, decocting liquid is filtered and merges and get final product.
The details of various aspects of the present invention will be able to detailed description in chapters and sections subsequently.By below and the description of claim, feature of the present invention, object and advantage will be more obvious.
Accompanying drawing explanation
Fig. 1 matched group testis: convoluted seminiferous tubule tube wall is without thickening, spermatogenic epithelium level is without increasing or reducing, the visible spermatogenic cell (HEX100) in different developmental phases;
Fig. 2 matched group testis: convoluted seminiferous tubule tube wall is without thickening, and basement membrane heavy wall is normal, spermatogenic epithelium level is without increasing or reducing, the visible spermatogenic cell (PASX100) in different developmental phases;
Fig. 3 matched group testis: convoluted seminiferous tubule tube wall is without thickening, spermatogenic epithelium level is without increasing or reducing, the visible spermatogenic cell (HEX100) in different developmental phases;
Fig. 4 matched group epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (PASX100);
The positive group of Fig. 5 testis: convoluted tubule of testis tube chamber is without subsiding, visible spermatogenic cells at different stages, interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
The positive group of Fig. 6 testis: convoluted tubule of testis tube wall basement membrane is without thickening, and spermatogenic cell quantity is many, and interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (PASX100);
The positive group of Fig. 7 epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
The positive group of Fig. 8 epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, without acute and chronic cell infiltration (PASX100);
Fig. 9 high dose group testis: convoluted tubule of testis tube wall level is thick, spermatogenic cell level is many, quantity is many, arrangement tight (black asterisk tubule), has volume mature sperm (green asterisk tubule) in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Figure 10 high dose group testis: convoluted seminiferous tubule thickness of pipe wall, spermatogenic cell level is many, quantity is many, dense arrangement.Convoluted seminiferous tubule basement membrane does not thicken (PASX100);
Figure 11 high dose group epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Figure 12 high dose group epididymis: epididymal duct intracavity is shown in volume sperm.Interstitial is without congested, edema, without acute and chronic cell infiltration (PASX100);
Dosage group testis in Figure 13: convoluted tubule of testis thickness of pipe wall, spermatogenic cell level is many, quantity is many, arrangement tight (black asterisk tubule).Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Dosage group testis in Figure 14: convoluted tubule of testis tube wall level is thick, spermatogenic cell level is many, quantity is many, arrangement is tight, and interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (PASX100);
Dosage group epididymis in Figure 15: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume mature sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Dosage group epididymis in Figure 16: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume mature sperm in tube chamber.Interstitial NIP, Interstitial cell is without hypertrophy (PASX100);
Figure 17 low dose group testis: convoluted tubule of testis tube wall level is thick, spermatogenic cell level is many, quantity is many, arrangement is tight, and interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Figure 18 low dose group testis: convoluted tubule of testis tube wall level is thick, spermatogenic cell level is many, quantity is many, arrangement is tight, and interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (PASX100);
Figure 19 low dose group epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (HEX100);
Figure 20 low dose group epididymis: epididymal duct epithelial cell, without degeneration, necrosis, is shown in volume sperm in tube chamber.Interstitial is without congested, edema, and without acute and chronic cell infiltration, Interstitial cell is without hypertrophy (PASX100).
The specific embodiment
The present invention's application Radix Astragali, Radix Morindae Officinalis, Herba Epimedii, QI invigorating kidney tonifying, supporing yang temperature fire.Wherein the Radix Astragali is solely heavy, benefiting qi and raising yang, warming middle-JIAO and strengthening the spleen; Radix Morindae Officinalis settling five organs, beneficial marrow, mends fire and not bright water, and two medicines are monarch drug altogether.The pungent temperature of Herba Epimedii, plays male sexuality and declines, except pains in the penis; Join Semen Cuscutae sweet flat, the kidney invigorating and essence nourishing, the use of reinforcement monarch drug QI invigorating warming YANG, is ministerial drug.Radix Polygoni Multiflori, Rhizoma Polygonati, nourishing the blood and yin, enriching kidney essence; Be slightly cold Radix Ophiopogonis, Yin-nourishing and body fluid promoting, and three medicines coordinate, and one helps the monarch and his subjects' YIN nourishing to mend essence, and the warm impairment of YIN blood that too consumes of two anti-the monarch and his subjects, is adjuvant drug.All medicines coordinate temper foot, essence and blood life, and the elder generation's yang-energy day after tomorrow must be mended, cloudy smart must filling, and the equilibrium between yin and yang of kidney, marrow benefit foot, amounts to warming and recuperating the spleen and kidney, the effect of YIN nourishing replenishing essence.
The Chinese crude drug Radix Astragali is the root of leguminous herbaceous plant's Radix Astagali, Radix Astragali, has the effects such as invigorating QI to consolidate the body surface resistance, diuretic detumescent, expelling pus and toxin by strengthening QI, granulation promoting.Modern pharmacological research confirms: the Radix Astragali can strengthen the phagocytic function of reticuloendothelial system, and blood leukocytes and multinuclear leucocyte quantity are being shown to be increased, and macrophage phagocytic percentage rate and phagocytic index is being shown and rise, and humoral immunization, cellular immunization are all had to facilitation.In addition the ability that, the Radix Astragali can also enhanced virus inducement interferon, promote organism metabolism, blood pressure lowering, protect the liver etc.
Be the tuber of liliaceous plant dwarf lilyturf Radix Ophiopogonis, cold nature, sweet in the mouth, micro-hardship.Can nourishing YIN and moistening the lung, the relieving restlessness that clears away heart-fire, reinforcing stomach reg fluid.Modern pharmacological research confirms: effectively reduce free radical Radix Ophiopogonis, increase SOD, stabilizing cell membrane, promotes vascular endothelial cell energy metabolism, regulates the secretory function of vascular endothelial cell.Radix Ophiopogonis extract can significantly reduce the aggregation rate of rat platelet, reduces blood viscosity, thus prevention of stroke.Radix Ophiopogonis, n-butanol extract can obviously reduce normal mouse blood sugar concentration, had dose dependent.Ophiopogonin and Radix Ophiopogonis polysaccharide can significantly increase mice organ thymus, spleen weight, and activate the phagocytic function of mice reticuloendothelial system, improve Hemolysin level, increase the immunity of body.
Radix Morindae Officinalis is the dry root of Maguireothamnus speciosus Radix Morindae Officinalis.Slightly warm in nature, acrid-sweet flavor, has kidney-replenishing, bone and muscle strengthening, the function of cold-dispelling.Modern pharmacological research confirms: Radix Morindae Officinalis has the immunologic function of adjusting, adjusting thyroid function, defying age, resisting fatigue, hypermnesis, antitumor, promotion bone growth, promotes the various active such as hemopoietic stem cell proliferation.
Herba Epimedii has another name called Herba Epimedii, is the dried leaves of Berberidaceae plant Herba Epimedii, arrow leaf Herba Epimedii, pubescence Herba Epimedii or Herba Epimedii.Acrid in the mouth, sweet, warm in nature.Can kidney invigorating and YANG supporting; Expelling wind and removing dampness; Strong muscle key bone.Motherland's medical science thinks, Herba Epimedii has the effect of " beneficial vital essence, hard muscles and bones are mended waist knee joint, heart tonifying power ".Modern medicine study also confirms, Herba Epimedii has the patrogenesis of adjusting, stops osteoporosis, regulates the functions such as immunity, antioxidation, defying age, antitumor, anti-hepatotoxin, vasodilator.
Radix Polygoni Multiflori is the dried root of polygonum multiflorum thunb, and its nature and flavor are bitter, sweet, puckery, tepor.Pharmacology analysis shows, Radix Polygoni Multiflori, containing anthraquinone analog compound, mainly contains chrysophanol and emodin, is secondly chrysophanic acid (product of processing are containing chrysophanic acid), physcione and Radix Et Rhizoma Rhei anthrone, starch, crude fat, lecithin etc.The pharmacological action of Radix Polygoni Multiflori mainly contains: (1) slow down aging; (2) blood fat reducing and study of anti-atherogenic effect, Radix Polygoni Multiflori decoct can significantly reduce the content of plasma cholesterol, triglyceride and beta lipoprotein, illustrates that Radix Polygoni Multiflori can reduce blood fat, delays atherosclerotic formation and development.(3) protect the liver, the tetrahydroxy ethylene-β-D-Glucose glycoside in the Radix Polygoni Multiflori is the effective ingredient protecting the liver, and it can prevent that fatty liver, liver function injury and liver LPO from raising, and reduces serum glutamic pyruvic transminase and glutamic oxaloacetic transaminase, GOT; (4) antibacterial, the Radix Polygoni Multiflori can suppress the growth of mycobacterium hominis, shigella flexneri in vitro, and its anthraquinone derivatives has inhibitory action in various degree to pathogen such as the antibacterials such as staphylococcus aureus, streptococcus, diphtheria corynebacterium, anthrax bacillus and influenza virus, funguses.
Semen Cuscutae is the ripe dry seed of parasitics MANCAO cuscuta japonica Choisy.Nature and flavor are sweet, warm in nature.Can be by kidney controlling nocturnal emission with astringent drugs, nourishing the liver to improve visual acuity, antidiarrheal, antiabortive.Modern pharmacological research confirms: Semen Cuscutae has tonifying YANG, increases hypothalamic-pituitary-ovarian short luteal function, heart tonifying, blood pressure lowering, delays the effects such as Cataractogenesis.
Rhizoma Polygonati is is the dry rhizome of liliaceous plant Yunnan Rhizoma Polygonati, Rhizoma Polygonati or Polygonatum cyrtonema Hua.Nature and flavor are sweet, flat.Can nourishing YIN and moistening the lung; Invigorating the spleen and replenishing QI, Zishen fill fine.Modern pharmacological research confirms: Rhizoma Polygonati has antibacterial, antiviral, resisting fatigue, antioxidation, slow down aging, stops blooding, resists myocardial ischemia, reduces the effects such as blood glucose.
Chinese medicine composition of the present invention, it is to make with the raw material of Chinese medicine of following weight proportion:
The Radix Astragali 30～600g, Radix Ophiopogonis 10～300g, Radix Morindae Officinalis 10～300g, Herba Epimedii 5～120g, Radix Polygoni Multiflori Preparata 20～600g, Semen Cuscutae 30～600g, Rhizoma Polygonati (processed) 20～500g.
The preferred weight proportioning of described raw material of Chinese medicine is:
The Radix Astragali 60～240g, Radix Ophiopogonis 30～150g, Radix Morindae Officinalis 30～150g, Herba Epimedii 9～50g, Radix Polygoni Multiflori Preparata 50～300g, Semen Cuscutae 60～300g, Rhizoma Polygonati (processed) 50～200g.
The most preferably weight proportion of described raw material of Chinese medicine is:
Radix Astragali 120g, Radix Ophiopogonis 60g, Radix Morindae Officinalis 60g, Herba Epimedii 20g, Radix Polygoni Multiflori Preparata 100g, Semen Cuscutae 160g, Rhizoma Polygonati (processed) 100g.
Chinese medicine composition of the present invention can be used separately, or adds appropriate amount of auxiliary materials to make various oral formulations as granule, tablet, capsule, mixture, powder etc. by the conventional method of this area.
Chinese medicine composition of the present invention can be prepared by the following method: get each crude drug and mix by said ratio, decocting 2～3 times, each 1～3 hour, add 4～20 times of amounts of water, decocting liquid is filtered and merged, standing, leaching supernatant adds sucrose, be concentrated into about 1000ml, add appropriate sodium benzoate, filter, add water to about 1000ml, stir evenly, subpackage, both.Also can add again appropriate amount of auxiliary materials, further make according to a conventional method other dosage forms.For example, add dextrin, spraying is dry makes into fine powder, through dry granulation granulation; Or add Sodium Hydroxymethyl Stalcs, micropowder silica gel, soluble starch, and lactose and magnesium stearate, mix homogeneously, with 95% ethanol granule processed; 80 ℃ of dry rear tablet agents; Add appropriate micropowder silica gel, after mixing, incapsulate; Or part fine powder adds citric acid, starch granulation, all the other fine powders add sodium bicarbonate, starch granulation, and two kinds of granules add after magnesium stearate, mix homogeneously, tabletting.
Through following pharmacodynamics test, confirm, Chinese medicine composition of the present invention has following beneficial effect:
(1) yang deficiency symptom is improved significantly, can improve animal to low temperature stress tolerance;
(2) gonad is had to good protective effect, and can maintain to a certain extent the function of gonad;
(3) can significantly increase sperm count and sperm day growing amount, but can not cause the hypertrophy of the relevant organ of reproductive system and body of gland, the testosterone of animal, rate of teratosperm and motility of sperm are also significantly improved, illustrate that it has gentle Spermatogenic action;
(4) can significantly promote the reproductive performance of animal;
(5) immune system of animal is had some improvement.
As can be seen here; Chinese medicine composition of the present invention can improve the yang deficiency symptom of animal on the whole; improve the immunocompetence of animal; by its gentle Spermatogenic action; when significantly increasing animal sperm number and sperm day growing amount; can not cause the hypertrophy of animal genitals and body of gland, on the contrary gonad be had to good protective effect.It is only by promoting the motor capacity of sperm and the medicine that film function is treated male infertility, so it has clinical expansion and using value that its wholistic therapy effect will obviously be better than existing those beyond doubt.
The experimentation of Chinese medicine composition of the present invention on the impact of mice model of yang asthenia
1. experiment material
1.1 animal
Clean level ICR male mice, body weight 18-22 gram, provides (laboratory animal production licence: SCXK(Soviet Union) 2007-0001 by Yangzhou University's comparative medicine center; Laboratory animal occupancy permit: SYXK(Soviet Union) 2007-0030).
1.2 medicines:
Compound recipe Formica fusca capsule, You Shiqiang Pharmaceutical Group Co.,Ltd provides, lot number: 20100609. by Chinese medicine ingredients such as Formica fusca, Herba Epimedii, Fructus Lycii, Fructus Cnidiis.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. entrusts Shanghai first line of a couplet pharmaceutcal corporation, Ltd to produce, Shanghai medicine word Z05060356 processed, lot number: 041226.The clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.
1.3 medicine preparations, dosage and grouping
(1) positive drug: compound recipe Formica fusca capsule, every dress 0.42g, 3 of people one time, 3 times on the one, people's one consumption per day is 3.78g/70kg people, being converted to mice is 0.01g/20g body weight. mice administration volume 0.2ml/20g body weight, medicinal liquid compound concentration is 0.01g/0.2ml=0.05g/ml, mice administration volume 0.2ml/20g body weight.
(2) the clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.By the day for human beings, take 50ml and calculate, its mice dose,equivalent is 0.13ml/20g body weight/day.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), gets 2 milliliters of extractum, adds 48ml distilled water and mixes and obtain clinical people's mixture.Four degree Refrigerator stores.Approximately can reach use in a week.
The heavy dose of group of Chinese medicine composition of the present invention: 2 times of dose,equivalents of mice administration are 0.26ml/20g body weight/day.
Dosage group in Chinese medicine composition of the present invention: mice administration dose,equivalent is 0.13ml/20g body weight/day.
Chinese medicine composition small dose group of the present invention: 0.5 times of dose,equivalent of mice administration is 0.065ml/20g body weight/day.
(3) hydrocortisone injection 20ml/ props up, and 100 milligrams every, concentration is 100mg/20ml=500mg/100ml=0.5g/100ml, is 0.5% hydrocortisone.Except Normal group, all the other each treated animal every day gives 0.5% hydrocortisone according to the intramuscular injection of 12.5ml/Kg=0.125ml/10g body weight and makes kidney-yang deficiency model.
1.4 instruments and reagent:
Sartorius electronic balance, German Sartorius company;
Mice swimming pool
1L beaker
Stopwatch
2. test method
2.1 experimental programs:
Choose 60 of ICR male mices, body weight 18-22g, is divided into 6 groups, 10 every group: Normal group; Kidney-yang deficiency model group; Compound recipe Formica fusca capsule sun property of medicine matched group; Dosage group in the heavy dose of group of Chinese medicine composition of the present invention, Chinese medicine composition of the present invention; Chinese medicine composition small dose group of the present invention.Except normal group animal, the next day of every treated animal, according to 12.5ml/Kg, give 0.5% hydrocortisone and make kidney-yang deficiency model.After model group injection hydrocortisone, give equal-volume distilled water gavage the same as normal group.After positive controls and each administration treated animal injection hydrocortisone, by group, give respectively relative medicine gavage, successive administration 30 days, after administration in the 30th day 1 hour, record each treated animal body weight, autonomic activities number and low temperature (four degree) swimming time in 10 minutes, result is carried out statistical procedures.
2.2 date processing: to all data, use Microsoft Excel software to carry out date processing, and statistic analysis result.
3. result
3.1 impacts of Chinese medicine composition of the present invention on Mouse Kidney model of yang asthenia autonomic activities and swimming time
By table 1 data, can be found out, the high, medium and low dosage group of Chinese medicine composition of the present invention all can significantly increase (p<0.01) yang deficiency mice autonomic activities number of times (in 10min); The high, medium and low dosage group of Chinese medicine composition of the present invention all can obviously extend swimming time under (p<0.01) mice cryogenic conditions, show that Chinese medicine composition of the present invention has obvious anti-yang deficiency effect, alleviate yang deficiency degree, improve mice to low temperature stress tolerance.
The impact of table 1 Chinese medicine composition of the present invention on model of yang asthenia mice autonomic activities and low temperature swimming time
With normal group comparison, * p<0.05, * * p<0.01; With model group comparison, #p<0.05, ##p<0.01
4. conclusion
To model of yang asthenia mice, after each dose application of Chinese medicine composition of the present invention, all can alleviate yang deficiency degree, there is statistical significant difference.After medication, the medicine of each dosage group all can significantly improve the autonomic activities number of times of mice, extends low temperature swimming time.High dose group can make low temperature swimming time approach normal group.Show that Chinese medicine composition of the present invention improves significantly to the yang deficiency symptom of mice, can improve mice to low temperature stress tolerance.
The experimentation of Chinese medicine composition of the present invention on the impact of rat castration kidney-yang deficiency model
1. experiment material
1.1 animals: SD rat, 40-60 gram
90 of SD male rats, in age in 4-5 week, body weight 40～60g, provides (laboratory animal production licence: SCXK(Zhejiang) 2008-0033 by Zhejiang Province's Experimental Animal Center; Laboratory animal occupancy permit: SYXK(Soviet Union) 2007-0030).
1.2 medicines:
Compound recipe Formica fusca capsule, You Shiqiang Pharmaceutical Group Co.,Ltd provides, lot number: 20100609. by Chinese medicine ingredients such as Formica fusca, Herba Epimedii, Fructus Lycii, Fructus Cnidiis.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. entrusts Shanghai first line of a couplet pharmaceutcal corporation, Ltd to produce, Shanghai medicine word Z05060356 processed, lot number: 041226.The clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.
1.3 medicine preparations, dosage and grouping
(1) positive drug: compound recipe Formica fusca capsule, every dress 0.42g, 3 of people one time, 3 times on the one, people's one consumption per day is 3.78g/70kg people, being converted to rat is 0.068g/200g body weight. rat administration volume 2ml/200g body weight, medicinal liquid compound concentration is 0.068g/2ml=0.034g/ml, rat administration volume 2ml/200g body weight.
(2) the clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.By the day for human beings, take 50ml and calculate, its rat dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), gets 2 milliliters of extractum, adds 48ml distilled water and mixes and obtain clinical people's mixture.Four degree Refrigerator stores.
The heavy dose of group of Chinese medicine composition of the present invention: 2 times of dose,equivalents of rat administration are 1.8ml/200g body weight/day.
Dosage group in Chinese medicine composition of the present invention: rat administration dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition small dose group of the present invention: 0.5 times of dose,equivalent of rat administration is 0.45ml/200g body weight/day.
1.4 instruments and reagent:
Sartorius electronic balance, German Sartorius company;
BECKMAIV-COULTER DXI800 testosterone detectable lot number: 020111
FITC anti-rat CD3 antibody, PE anti-rat CD8 antibody, PE anti-rat CD4 antibody is provided by BIOLEGEND company.
BD FACSCalibur flow cytometer
2. test method
2.1 experimental programs:
Laboratory animal is divided into 6 groups at random, i.e. Normal group, model group, positive drug (compound recipe Formica fusca capsule) group, dosage group, Chinese medicine composition low dose group of the present invention in Chinese medicine composition high dose group of the present invention, Chinese medicine composition of the present invention.12 every group of rats.Normal group; Model group; Sun property of medicine matched group; Dosage group in the heavy dose of group of Chinese medicine composition of the present invention, Chinese medicine composition of the present invention; Chinese medicine composition small dose group of the present invention.Except normal group animal, all rat Testectomy castrations, sew up the incision.Random packet after operation second day, is divided into model group and administration group.Model group and normal group give same volume distilled water gavage, and administration group gives respectively positive control medicine and tested medicine according to group.Successive administration 30 days, once a day, 24 hours femoral artery sacrificed by exsanguination rats after last administration, collect rat blood and also carry out anticoagulant, minute get 1ml centrifugal separation plasma after standing a period of time, and plasma sample detects the testosterone concentration in blood plasma.The rat of putting to death is dissected, and cuts the internal organs such as prostate, seminal fluid capsule, levator ani of animal, weighs, and calculates organ index.
2.2 date processing: to all data, use Microsoft Excel software to carry out date processing, and statistic analysis result.
3. result
The impact of 3.1 Chinese medicine compositions of the present invention on rat castration kidney-yang deficiency model organ coefficient
As can be seen from Table 2, compare with normal group, model group rat prostate and levator ani organ coefficient have significant difference (p<0.01), show rat castration kidney-yang deficiency model modeling success.With model group comparison, Chinese medicine composition of the present invention high, medium and low dosage group prostate and levator ani coefficient have significant difference (p<0.01), show after Chinese medicine composition administration of the present invention can significance to alleviate insufficiency of kidney-YANG degree after rat castration.
Table 2 Chinese medicine composition of the present invention on the impact of rat castration kidney-yang deficiency model organ coefficient (
n=10)
Note: with normal group comparison, * * p<0.01; With model group comparison, ##p<0.01;
3.2. the impact of Chinese medicine composition of the present invention on rat castration kidney-yang deficiency model blood parameters:
As can be seen from Table 3, compare with normal group, in castrated rats body, testosterone levels obviously declines, Chinese medicine composition high dose of the present invention (p<0.01) and middle dosage, low dose group (p<0.05) all can improve testosterone levels in castration male rat body by significance, and visible medicine can maintain the testosterone levels in rat body to a certain extent.
Table 3 Chinese medicine composition of the present invention on castration kidney-yang deficiency model rat TESTO and immune impact (
n=10)
Note: with model group comparison, * p<0.05, * * p<0.01.With the comparison of blank group, #p<0.05.
4. conclusion
To rat castration kidney-yang deficiency model, after each dose application of Chinese medicine composition of the present invention, all can alleviate insufficiency of kidney-YANG degree, there is statistical significant difference.Chinese medicine composition of the present invention can maintain the testosterone concentration in rat body simultaneously.After medication, the medicine of each dosage group all can significantly improve prostate coefficient and the levator ani coefficient of rat, shows that Chinese medicine composition of the present invention has good protective effect to the gonad of male rat, and can maintain to a certain extent the function of gonad.
The impact research of Chinese medicine composition of the present invention on rat spermatozoa vigor and form
1. experiment material
1.1 animals:
90 of SD male rats, in age in 4-5 week, body weight 40～60g, provides (laboratory animal production licence: 2007-012 SCXK(army) by Nanking Military Area Command of the Chinese People's Liberation Army's medical animal experiment center; Laboratory animal occupancy permit: SYXK(Soviet Union) 2007-0030).
1.2 medicines:
Compound recipe Formica fusca capsule, You Shiqiang Pharmaceutical Group Co.,Ltd provides, lot number: 20100609. by Chinese medicine ingredients such as Formica fusca, Herba Epimedii, Fructus Lycii, Fructus Cnidiis.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. entrusts Shanghai first line of a couplet pharmaceutcal corporation, Ltd to produce, Shanghai medicine word Z05060356 processed, lot number: 041226.The clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.
1.3 medicine preparations, dosage and grouping
(1) positive drug: compound recipe Formica fusca capsule, every dress 0.42g, 3 of people one time, 3 times on the one, people's one consumption per day is 3.78g/70kg people, being converted to rat is 0.068g/200g body weight. rat administration volume 2ml/200g body weight, medicinal liquid compound concentration is 0.068g/2ml=0.034g/ml, rat administration volume 2ml/200g body weight.
(2) the clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.By the day for human beings, take 50ml and calculate, its rat dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), gets 2 milliliters of extractum, adds 48ml distilled water and mixes and obtain clinical people's mixture.Four degree Refrigerator stores.
The heavy dose of group of Chinese medicine composition of the present invention: 2 times of dose,equivalents of rat administration are 1.8ml/200g body weight/day.
Dosage group in Chinese medicine composition of the present invention: rat administration dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition small dose group of the present invention: 0.5 times of dose,equivalent of rat administration is 0.45ml/200g body weight/day.
1.4 reagent
BECKMAIV-COULTER DXI800 testosterone reagent lot number: 020111
Glucose ringer solution, laboratory autogamy
1.5 instruments:
Sartorius electronic balance, German Sartorius company;
2. test method
2.1 experimental programs:
Laboratory animal is divided into 5 groups at random, i.e. Normal group, positive drug (compound recipe Formica fusca capsule) group, dosage group, Chinese medicine composition low dose group of the present invention in Chinese medicine composition high dose group of the present invention, Chinese medicine composition of the present invention.10 every group of rats.By group, give respectively rat oral gavage administration (1ml/100g), normal group gives same volume distilled water (1ml/100g).Successive administration 35 days, once a day, 24 hours femoral artery sacrificed by exsanguination rats after last administration, collect rat blood and also carry out anticoagulant, minute get 1ml centrifugal separation plasma after standing a period of time, and plasma sample censorship detects the testosterone concentration in blood plasma; The rat of putting to death is dissected, and cuts the internal organs such as prostate, seminal fluid capsule, levator ani of animal, weighs, and calculates organ index.Get a side epididymis and be placed in 20ml beaker, add 0.1% glucose ringer solution 2ml, suitably shred, jolting, makes sperm automatic moving to solution at 30 degree insulation 10min.Get 1 and put on microscope slide, under low power microscope, observe the mobility of sperm, add up respectively percentage rate and the head of sperm simultaneously.The percentage rate of tail fracture sperm.And count sperm count on red, numeration of leukocyte plate.It is fixing that opposite side epididymis and testis are placed in formalin, makes tissue slice, and in microscopy epididymal and epididymis, whether sperm receives damage and morphologic change; Whether microscopy testis tissue is normal.
2.2 date processing:
To all data, use Microsoft Excel software to carry out date processing, and statistic analysis result.
3. result
3.1 impacts of Chinese medicine composition of the present invention on male rat organ coefficient
Each dosage group of Chinese medicine composition of the present invention, on the not significant impact of the prostate of normal male rat, seminal vesicle fluid, testis, epididymis, preputial glands, can not cause the hypertrophy of these organs as seen from the data in Table 4.
Table 4 Chinese medicine composition of the present invention on the impact of male rat organ coefficient (
n=10)
With normal group comparison, * P<0.05; With the comparison of positive drug group, #P<0.01.
3.2. the impact of Chinese medicine composition of the present invention on rat spermatozoa vigor:
As seen from the data in Table 5 each dosage group of Chinese medicine composition of the present invention to can significantly improve normal male rat spermatozoa number and sperm day growing amount, there is the trend that improves testosterone levels in male rat body, but compare no difference of science of statistics with normal group, but do not cause that rate of teratosperm improves.Prompting medicine has gentle Spermatogenic action and without causing the side effect of sperm deformity.
Table 5 Chinese medicine composition of the present invention on the impact of male rat testosterone and motility of sperm (
n=10)
With normal group comparison, * * P<0.01; With the comparison of positive drug group, ##P<0.01.
4. conclusion
Chinese medicine composition of the present invention can significantly increase male rat sperm count and sperm day growing amount, but can not cause the hypertrophy of the relevant organ of Reproductive System in Male Rats and body of gland, to the also obviously improvement effect of thing of the testosterone of rat, rate of teratosperm and motility of sperm, prompting medicine has gentle Spermatogenic action.
The histopathological study of Chinese medicine composition of the present invention on the impact of rat testicle epididymis
1. materials and methods:
1.1. laboratory animal and grouping:
50 of rats, 5 groups, 10 every group.
Group: normal group, positive group (compound recipe Formica fusca capsule), Chinese medicine composition high dose group of the present invention, dosage group in Chinese medicine composition of the present invention, Chinese medicine composition low dose group of the present invention.
1.2. send and pick up internal organs: testis and epididymis.
1.3. method: after testis and epididymis are in vitro, put into immediately 10% formalin solution fixing, routine is drawn materials, dehydration, paraffin embedding film-making, slice thick 4 μ m, HE dyeing and PAS dye, the morphological change of om observation testis and epididymis.Testis: convoluted seminiferous tubule cell arrangement has or not disorder, basement membrane to have or not to thicken, and spermatogenesis phenomenon has unchanged (increase or reduce); Interstitial has or not edema, and Interstitial cell has or not hypertrophy.Epididymis: epididymal duct epithelial cell has or not degeneration, necrosis, has or not sperm, interstitial to have or not hyperemia, edema, cell infiltration in tube chamber.According to pathological changes light and heavy degree, sxemiquantitative is slight " ± " successively, slight "+", moderate " ++ ", severe " +++ ", is "-" without pathological changes tissue mark.
2. result
2.1 normal group (10):
2.1.1. testis: normal rat testis is comprised of convoluted seminiferous tubule, the spermatogenic cell hierarchal arrangement of different developmental phases, from the basic unit of convoluted seminiferous tubule to tube chamber, be followed successively by spermatogonium, primary spermatocyte, secondary spermatocyte, spermatid, with the sperm of chamber face maturation, sustenticular cell is distributed between each phase spermatogenic cell.Between convoluted seminiferous tubule, for being rich in blood vessel and vasculolymphatic loose connective tissue, be wherein also shown in a small amount of Interstitial cell.(Fig. 1)
The basement membrane that dye in the peripheral Yihong of visible convoluted seminiferous tubule of PAS when dyeing, basement membrane is without thickening, spermatid no abnormality seen.(Fig. 2)
2.1.2. epididymis: epididymal duct epithelial cell is column or cubic, without degeneration, necrosis, the pathological changes such as come off, the visible volume mature sperm of intracavity.Interstitial has no vasodilation hyperemia, hemorrhage, cell infiltration.(Fig. 3)
The basement membrane that dye in the peripheral Yihong of visible epididymal duct of PAS when dyeing, basement membrane is without thickening, sperm no abnormality seen in tube chamber.(Fig. 4)
2.2. positive group (recurrence Formica fusca capsule) (10):
2.2.1. testis: testis by convoluted seminiferous tubule structure without extremely, the spermatogenic cell hierarchal arrangement of different developmental phases is orderly, from the basic unit of convoluted seminiferous tubule to tube chamber, be followed successively by spermatogonium, primary spermatocyte, secondary spermatocyte, spermatid, with the sperm of chamber face maturation, sustenticular cell is distributed between each phase spermatogenic cell.Between convoluted seminiferous tubule, for being rich in blood vessel and vasculolymphatic loose connective tissue, be wherein also shown in a small amount of Interstitial cell, 2 slight edema of interstitial.(Fig. 5)
The basement membrane that dye in the peripheral Yihong of visible convoluted seminiferous tubule of PAS when dyeing, basement membrane is without thickening, spermatid no abnormality seen.(Fig. 6)
2.2.2. epididymis: epididymal duct epithelial cell is column or cubic, without degeneration, necrosis, the pathological changes such as come off, the visible volume mature sperm of intracavity.Interstitial has no vasodilation hyperemia, hemorrhage, cell infiltration.(Fig. 7)
The basement membrane that dye in the peripheral Yihong of visible epididymal duct of PAS when dyeing, basement membrane is without thickening, sperm no abnormality seen in tube chamber.(Fig. 8)
2.3 Chinese medicine composition high dose group of the present invention (10):
2.3.1. testis: testes histological structure is identical with normal group, convoluted seminiferous tubule without obviously atrophy, subside or expand, each layer of spermatogenic cell is without obviously reducing or increasing, interstitial is without obvious hyperemia, without cell infiltration, 3 interstitials have slight edema.(Fig. 9)
The basement membrane that during PAS dyeing, dye in the peripheral Yihong of visible convoluted seminiferous tubule, basement membrane is without thickening, and spermatid quantity shows no obvious abnormalities.(Figure 10)
2.3.2. epididymis: epididymal duct epithelial cell is column or cubic, without degeneration, necrosis, the pathological changes such as come off, the visible volume mature sperm of intracavity, has no degeneration, non-viable non-apoptotic cell increases.Interstitial blood vessel is without dilatation and congestion, hemorrhage, and 7 without cell infiltration, and 1 interstitial has slight cell infiltration, is mainly mononuclear phagocyte and lymphocyte.(Figure 11)
Basement membrane, the nothing that during PAS dyeing, dye in the peripheral Yihong of epididymal duct thicken, and in tube chamber, sperm shows no obvious abnormalities.(Figure 12)
2.4. dosage group (10) in Chinese medicine composition of the present invention:
2.4.1. testis: testes histological structure is identical with normal group, convoluted seminiferous tubule without obviously atrophy, subside or expand, each layer of spermatogenic cell be without obviously reducing or increasing, interstitial is without obvious hyperemia, without cell infiltration, 3 slight edema of interstitial.(Figure 13)
The basement membrane that dye in the peripheral Yihong of visible convoluted seminiferous tubule of PAS when dyeing, basement membrane is without thickening, spermatid no abnormality seen.(Figure 14)
2.4.2. epididymis: epididymal duct epithelial cell is column or cubic, without degeneration, necrosis, the pathological changes such as come off, the visible volume mature sperm of intracavity, has no volume degeneration, downright bad cell.Interstitial blood vessel is without dilatation and congestion, hemorrhage, without cell infiltration.(Figure 15)
Basement membrane, the nothing that during PAS dyeing, dye in the peripheral Yihong of epididymal duct thicken, and in tube chamber, sperm shows no obvious abnormalities.(Figure 16)
2.5 Chinese medicine composition low dose group of the present invention (10):
2.5.1. testis: testes histological structure is identical with normal group, convoluted seminiferous tubule without obviously atrophy, subside or expand, each layer of spermatogenic cell is without obviously reducing or increasing, interstitial is without hyperemia, edema and cell infiltration.(Figure 17)
The basement membrane that dye in the peripheral Yihong of visible convoluted seminiferous tubule of PAS when dyeing, basement membrane is without thickening, spermatid no abnormality seen.(Figure 18)
2.5.2. epididymis: epididymal duct epithelial cell is column or cubic, without degeneration, necrosis, the pathological changes such as come off, the visible volume mature sperm of intracavity, has no volume degeneration, downright bad cell.Interstitial blood vessel is without dilatation and congestion, hemorrhage, without cell infiltration.(Figure 19)
Basement membrane, the nothing that during PAS dyeing, dye in the peripheral Yihong of epididymal duct thicken, and in tube chamber, sperm shows no obvious abnormalities.(Figure 20)
3. conclusion
Convoluted seminiferous tubule is without obviously expansion or atrophy in HE and PAS stained for the high, medium and low dosage rat testicle of Chinese medicine composition of the present invention and epididymis, and basement membrane is without thickening.Interstitial cell is without hypertrophy or obviously minimizing, and nothing is acute, chronic inflammatory reaction significantly.
The experimentation of Chinese medicine composition of the present invention on the impact of male mice reproductive function
1. experiment material
1.1 animal
Kunming mouse, 40 of bull mices, body weight 20-25 gram; 480 of adult female mices, body weight 20-25 gram.By Zhejiang Province's Experimental Animal Center, provide (laboratory animal production licence: SCXK(Zhejiang) 2008-0033; Laboratory animal occupancy permit: SYXK(Soviet Union) 2007-0030).
1.2 medicines:
Compound recipe Formica fusca capsule, You Shiqiang Pharmaceutical Group Co.,Ltd provides, lot number: 20100609. by Chinese medicine ingredients such as Formica fusca, Herba Epimedii, Fructus Lycii, Fructus Cnidiis.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. entrusts Shanghai first line of a couplet pharmaceutcal corporation, Ltd to produce, Shanghai medicine word Z05060356 processed, lot number: 041226.The clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.
1.3 medicine preparations, dosage and grouping
(1) positive drug: compound recipe Formica fusca capsule, every dress 0.42g, 3 of people one time, 3 times on the one, people's one consumption per day is 3.78g/70kg people, being converted to mice is 0.01g/20g body weight. mice administration volume 0.2ml/20g body weight, medicinal liquid compound concentration is 0.01g/0.2ml=0.05g/ml, mice administration volume 0.2ml/20g body weight.。
(2) the clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.By the day for human beings, take 50ml and calculate, its mice dose,equivalent is 0.13ml/20g body weight/day.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), gets 2 milliliters of extractum, adds 48ml distilled water and mixes and obtain clinical people's mixture.Four degree Refrigerator stores.Approximately can reach use in a week.
The heavy dose of group of Chinese medicine composition of the present invention: 2 times of dose,equivalents of mice administration are 0.26ml/20g body weight/day.
Dosage group in Chinese medicine composition of the present invention: mice administration dose,equivalent is 0.13ml/20g body weight/day.
Chinese medicine composition small dose group of the present invention: 0.5 times of dose,equivalent of mice administration is 0.065ml/20g body weight/day.
1.4 instruments and reagent:
Sartorius electronic balance, German Sartorius company;
2. test method
2.1 experimental programs:
Choose health, maturation, body weight the mice of 20-25g, 40 of male mices, are divided into 5 groups at random, 8 every group; 400 of female mices, are divided into 5 groups at random, 80 every group.Male mice gives respectively distilled water, positive drug, the high, medium and low dosage of Chinese medicine composition of the present invention by group.Successive administration 30 days, once a day, in after administration the 23rd day, 1 of male mice is put in 10 Female Rats groups with reproductive performance, and later every morning is used normal saline flushing mouse vagina, and person is the Mus of becoming pregnant to detect sperm, examine altogether 7 days, record the Mus number of becoming pregnant, settle accounts average pregnancy rate, add up the difference between each group.
2.2 date processing:
To all data, use Microsoft Excel software to carry out date processing, and statistic analysis result.
3. result
3.1 impacts of Chinese medicine composition of the present invention on male mice reproductive function
By table 6 data, can be found out, Chinese medicine composition object height of the present invention, middle dosage group all can significantly increase the fertilization success rate that bull mice makes female mice, the average pregnancy rate of female mice significantly improves (p<0.01), shows that Chinese medicine composition of the present invention has the effect of obvious enhancing male mice reproductive function.
The impact of table 6 Chinese medicine composition of the present invention on male mice reproductive function
With normal group comparison, * p<0.05, * * p<0.01;
4. conclusion
After Chinese medicine composition object height of the present invention, middle dose application, all can increase the pregnancy rate of female mice, there is statistical significant difference.After medicine taking prompting, the male mice reproductive function compared with normal group male mice of high, middle dosage group is significantly increased, and medicine can have the reproductive performance of remarkable lifting male mice.
The impact that the immune functions of rats that Chinese medicine composition of the present invention causes cyclophosphamide suppresses
1. experiment material
1.1 animals:
90 of SD male rats, body weight 90-180g, provides (laboratory animal production licence: SCXK(Zhejiang) 2008-0033 by Zhejiang Province's Experimental Animal Center; Laboratory animal occupancy permit: SYXK(Soviet Union) 2007-0030).
1.2 medicines:
Compound recipe Formica fusca capsule, You Shiqiang Pharmaceutical Group Co.,Ltd provides, lot number: 20100609. by Chinese medicine ingredients such as Formica fusca, Herba Epimedii, Fructus Lycii, Fructus Cnidiis.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), Renji Hospital Attached to Medical College of Shanghai Jiaotong Univ. entrusts Shanghai first line of a couplet pharmaceutcal corporation, Ltd to produce, Shanghai medicine word Z05060356 processed, lot number: 041226.The clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.
1.3 medicine preparations, dosage and grouping
(1) positive drug: compound recipe Formica fusca capsule, every dress 0.42g, 3 of people one time, 3 times on the one, people's one consumption per day is 3.78g/70kg people, being converted to rat is 0.068g/200g body weight. rat administration volume 2ml/200g body weight, medicinal liquid compound concentration is 0.068g/2ml=0.034g/ml, rat administration volume 2ml/200g body weight.
(2) the clinical people's using method of Chinese medicine composition of the present invention is oral, one time 25 milliliters, 2-3 time on the one.By the day for human beings, take 50ml and calculate, its rat dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition extractum of the present invention (25 times of concentrated solutions), gets 2 milliliters of extractum, adds 48ml distilled water and mixes and obtain clinical people's mixture.Four degree Refrigerator stores.
The heavy dose of group of Chinese medicine composition of the present invention: 2 times of dose,equivalents of rat administration are 1.8ml/200g body weight/day.
Dosage group in Chinese medicine composition of the present invention: rat administration dose,equivalent is 0.9ml/200g body weight/day.
Chinese medicine composition small dose group of the present invention: 0.5 times of dose,equivalent of rat administration is 0.45ml/200g body weight/day.
1.4 instruments and reagent:
Sartorius electronic balance, German Sartorius company;
FITC anti-rat CD3 antibody, PE anti-rat CD8 antibody, PE anti-rat CD4 antibody is provided by BIOLEGEND company.
BD FACSCalibur flow cytometer
2. test method
2.1 experimental programs:
Laboratory animal is divided into 6 groups at random, i.e. Normal group, model group, positive drug (compound recipe Formica fusca capsule) group, dosage group, Chinese medicine composition low dose group of the present invention in Chinese medicine composition high dose group of the present invention, Chinese medicine composition of the present invention.10 every group of rats.Except normal group animal, all rat single intraperitoneal injection cyclophosphamide random packet after 75mg/kg days, is divided into model group and each administration group.Model group and normal group give same volume distilled water gavage, and administration group gives respectively positive control medicine and tested medicine according to group.Successive administration 30 days, once a day, 24 hours femoral artery sacrificed by exsanguination rats after last administration, collect rat blood and carry out anticoagulant, and taking heparin anticoagulation 4ml, is placed in centrifugal 2000rpm on lymphocyte separation medium, 20min.The PBM that draws interface, Hanks liquid is washed once, centrifugal 1000rpm, 10min.Add and dissolve people RBC liquid, 37 degree temperature are incubated 5min, the centrifugal 10min of 1000rpm.Hanks liquid is washed once, gets PBM1 * 106 cell/pipe, adds respectively fluorescein-labeled CD
3, CD
4, CD
8monoclonal antibody or simultaneously add different fluorescein-labeled antibody and antibody diluent 100 μ l, the lower standing 3min of lucifuge of 4 degree.Hanks liquid washed twice, the centrifugal 4min of 2000rpm, after 30-40 μ m nylon membrane filters, flow cytometer detects.
2.2 date processing:
To all data, use Microsoft Excel software to carry out date processing, and statistic analysis result.
3. result
The impact that the immune functions of rats that 3.1 Chinese medicine compositions of the present invention cause cyclophosphamide suppresses
As can be seen from Table 7, cyclophosphamide can obviously suppress rat function, shows as rat CD3, CD4, and cd8 cell level declines, the Chinese medicine composition high dose medicament of the present invention CD in rat body that can significantly raise
3, CD4, cd8 cell level, in Chinese medicine composition of the present invention, low-dose drugs is for CD
3, CD
4, CD
8expression all there is no the impact of significance, show that Chinese medicine composition of the present invention has certain improvement effect to the immune system of rat.
The impact that the immune functions of rats that table 7 Chinese medicine composition of the present invention causes cyclophosphamide suppresses (
n=10)
Note: with model group comparison, * p<0.05, * * p<0.01.With the comparison of blank group, ##p<0.01.
4. conclusion
The immune functions of rats inhibition that rat cyclophosphamide is caused, can alleviate the downtrod degree of immunologic function after Chinese medicine composition high dose application of the present invention, has statistical significant difference.Show that Chinese medicine composition of the present invention has certain improvement effect to the immune system of rat.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percent, ratio, ratio or umber by weight.
Unless otherwise defined, the same meaning that all specialties of using in literary composition and scientific words and one skilled in the art are familiar.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The use that better implementation method described in literary composition and material only present a demonstration.
The above-mentioned feature that the present invention mentions, or the feature that embodiment mentions can combination in any.All features that patent specification discloses can with any composition forms use, each feature disclosing in description, can anyly provide the alternative characteristics of identical, impartial or similar object to replace.Therefore apart from special instruction, the feature disclosing is only the general example of equalization or similar features.
Embodiment 1
Material Radix Astragali 30g, Radix Ophiopogonis 10g, Radix Morindae Officinalis 100g, Herba Epimedii 120g, Radix Polygoni Multiflori Preparata 600g, Semen Cuscutae 600g, Rhizoma Polygonati (processed) 500g get it filled.Decoct with water 2 times, add for the first time 12 times of amounts of water (weight ratio) and decoct 2 hours, add for the second time 6 times of amounts of water (weight ratio) and decoct 1.5 hours, decocting liquid filters, and merges, and is concentrated into relative density 1.35, adds appropriate lactose, and granulation 1000g, obtains.
Embodiment 2
Radix Astragali 240g, Radix Ophiopogonis 150g, Radix Morindae Officinalis 30g, Herba Epimedii 50g, Radix Polygoni Multiflori Preparata 50g, Semen Cuscutae 60g, Rhizoma Polygonati (processed) 50g.Decoct with water 3 times, adding for the first time 10 times of amounts of water (weight ratio) decocts 1 hour, adding for the second time 8 times of amounts of water (weight ratio) decocts 1 hour, add for the third time 4 times of amounts of water (weight ratio) and decoct 0.5 hour, decocting liquid filters, and merges, concentrating under reduced pressure is dry, add appropriate amount of starch, lactose, magnesium stearate, make 1000, tablet, obtain.
Embodiment 3
Radix Astragali 600g, Radix Ophiopogonis, 30g, Radix Morindae Officinalis 10g, Herba Epimedii 9g, Radix Polygoni Multiflori Preparata 20g, Semen Cuscutae 30g, Rhizoma Polygonati (processed) 20g, decocted with water 3 times, adds for the first time 12 times of amounts of water (weight ratio) and decoct 2 hours, add for the second time 10 times of amounts of water (weight ratio) and decoct 1 hour, decocting liquid filters, and merges, standing 6～8 hours, leaching supernatant added sucrose 85g, was concentrated into about 1000ml, add sodium benzoate 3g, filter, add water to 1000ml, stir evenly, subpackage, obtains.
Embodiment 4
Radix Astragali 60g, Radix Ophiopogonis, 300g, Radix Morindae Officinalis 300g, Herba Epimedii 5g, Radix Polygoni Multiflori Preparata 300g, Semen Cuscutae 300g, Rhizoma Polygonati (processed) 200g, decocted with water 2 times, adds for the first time 8 times of amounts of water (weight ratio) and decoct 1 hour, add for the second time 6 times of amounts of water (weight ratio) and decoct 1 hour, decocting liquid filters, and merges, standing 6～8 hours, leaching supernatant, concentrate drying, pulverize, add appropriate amount of starch, lactose, mix, encapsulated, make 1000, obtain.
Embodiment 5
Radix Astragali 120g, Radix Ophiopogonis, 60g, Radix Morindae Officinalis 60g, Herba Epimedii 20g, Radix Polygoni Multiflori Preparata 100g, Semen Cuscutae 160g, Rhizoma Polygonati (processed) 100g, decocted with water 2 times, adds for the first time 12 times of amounts of water (weight ratio) and decoct 2 hours, add for the second time 10 times of amounts of water (weight ratio) and decoct 1 hour, decocting liquid filters, and merges, standing 6～8 hours, leaching supernatant added sucrose 85g, was concentrated into about 1000ml, add sodium benzoate 3g, filter, add water to 1000ml, stir evenly, subpackage, obtains.
Embodiment 6
Get the prepared oral liquid of embodiment 5, treatment clinical diagnosis is patients with infertility 30 examples, and therapeutic scheme and result are as follows:
Administering mode: oral, one day 3 times, a 20ml.
The course for the treatment of: 24 weeks.
Curative effect determinate standard: compare with last semen analysis result before curative effect judges to treat.
Cure: sperm concentration >20 * 10 in semen analysis
6mL
-1, and a+b level sperm ratio >50%, or a level sperm ratio >25%, or during treatment, spouse is conceived;
Effective: sperm concentration improves >50%, and the more former radix of a+b level sperm ratio improves >50% or the more former radix of a level sperm ratio improves >50%;
Effective: do not reach criterion of cure, sperm concentration improves >25%, and the more former radix raising 25% of a+b level sperm ratio or the more former radix raising 25% of a level sperm ratio;
Invalid: do not reach criterion of cure, sperm concentration improves <25%, the more former radix of a+b level sperm ratio improves <25% or the more former radix of a level sperm ratio improves <25%.
Therapeutic outcome: cure 10 examples, account for 33.33%; Effective 5 examples, account for 16.67%; Effective 4 examples, account for 13.33%; Invalid 11 examples, account for 36.67%; Total effective rate is 63.33%.
Many aspects involved in the present invention have been done as above and have been set forth.Yet, it should be understood that before not departing from spirit of the present invention and put, those skilled in the art can be equal to and change and modify it, and described change and modification fall into the coverage of the application's claims equally.
Claims (3)
1. a Chinese medicine composition for the treatment of deficiency of spleen-YANG and kidneyYANG oligospermia, is characterized in that, it is to make with the raw material of Chinese medicine of following weight proportion:
Radix Astragali 120g, Radix Ophiopogonis 60g, Radix Morindae Officinalis 60g, Herba Epimedii 20g, Radix Polygoni Multiflori Preparata 100g, Semen Cuscutae 160g, Rhizoma Polygonati (processed) 100g.
2. the application of Chinese medicine composition as claimed in claim 1 in the medicine of preparation treatment deficiency of spleen-YANG and kidneyYANG oligospermia.
3. the preparation method of Chinese medicine composition as claimed in claim 1, is characterized in that, it comprises the following steps: to get each crude drug mixes, decocting twice, and each 1～3 hour, add 4～20 times of amounts of water, decocting liquid is filtered and merges and get final product.
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| CN108815379A (en) * | 2018-08-31 | 2018-11-16 | 王志强 | A kind of promotion raw ovum or the Chinese medicine of production of sperm and preparation method thereof |
| CN110538251A (en) * | 2019-09-25 | 2019-12-06 | 李明杰 | Traditional Chinese medicine composition for treating prostate diseases and preparation method and application thereof |
| CN112237607A (en) * | 2020-11-11 | 2021-01-19 | 武汉林宝莱生物科技有限公司 | Formula and preparation process of epimedium carbene tablet preparation |
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| CN103006930A (en) * | 2012-11-30 | 2013-04-03 | 重庆市中药研究院 | Application of ginseng and rhizoma curculiginis combined medicinal herbs in preparation of medicament for treating male infertility |
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2013
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103006930A (en) * | 2012-11-30 | 2013-04-03 | 重庆市中药研究院 | Application of ginseng and rhizoma curculiginis combined medicinal herbs in preparation of medicament for treating male infertility |
Non-Patent Citations (2)
| Title |
|---|
| 王鸿祥等.黄麦合剂治疗脾肾阳虚少和弱精症33例.《医药导报》.2011,第30卷(第01期),24-28. * |
| 黄麦合剂治疗脾肾阳虚少和弱精症33例;王鸿祥等;《医药导报》;20110131;第30卷(第01期);24-28 * |
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