CN1457799A - Compound composition for curing male's sterility - Google Patents
Compound composition for curing male's sterility Download PDFInfo
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- CN1457799A CN1457799A CN 02115233 CN02115233A CN1457799A CN 1457799 A CN1457799 A CN 1457799A CN 02115233 CN02115233 CN 02115233 CN 02115233 A CN02115233 A CN 02115233A CN 1457799 A CN1457799 A CN 1457799A
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Abstract
The compound composition for treating male's sterility consists of mainly arginine and zinc sulfate in the ratio of 4.62-19.24. It can raise the quantity and quality of sperm, raise the activity of sperm and reduce the deformity rate of sperm, has less toxicity. Compared with single arginine and zinc sulfate, the present invention has even higher treating effect.
Description
1, technical field:
The present invention relates to a kind of compound that is used for the treatment of male sterility, relate in particular to a kind of compound that is used for the treatment of male sterility by arginine and zinc sulfate combination.
2, background technology:
Sterile is a global medical problem.Refuse World Health Organization's survey data, have 15% the couple at child-bearing age to have sterile problem, the whole world has about 80,000,000 pairs, and also increase progressively with 2,000,000 pairs every year.The relevant bibliographical information of China, the infertile couples incidence rate accounts for the couple at child-bearing age's 12.5%.Infertile couples is often painful for having no children, and mental pressure and social pressure are very big.Spend to seek medical advice and medicine for asking children, not stinting, even mistake believes itinerant doctor, bad doctor, postpone treatment opportunity.Research and production are used for the treatment of the medicine of male sterility, are needs of society, are patient's serious hopes, have far-reaching social meaning and realistic meaning.
Though the reason of male sterility is complicated, but it is the most outstanding and general with sperm number deficiency, sperm quality difference, the medicine of research at present and production raising mankind spermatozoon number and quality is smaller, free amino acid participates in the metabolism of sperm in the human refining, be sperm energy supplement source, can promote the activity of sperm.Aminoacid has chelation again, can sperm be shielded in conjunction with deleterious metal ion.Aminoacid also participates in the synthetic of hormone enzyme and other active substances.Influence the synthetic and renewal of above-mentioned substance if aminoacid is not enough, certainly will cause the spermatogenesis obstacle.Studies confirm that recently, arginine has stimulates hypophysis to discharge the effect of promoting sexual gland hormone, oral arginine not only can improve the quantity of sperm, and can improve the quality of sperm, thus can be used as the treatment semen volume not enough and less, the male sterility due to weak, the teratozoospermia.Therefore now on the market also by the folk prescription medicine that with the arginine is main component, but therapeutic effect is undesirable.
3, summary of the invention:
The purpose of this invention is to provide a kind of compound that is used for the treatment of male sterility, it adds other effective ingredient composition compound on the folk prescription medicine basis that with the arginine is main component originally, further improve the quality and quantity of sperm, reach the effect of treatment preferably.
The present invention is achieved by the following technical solution: it mainly is made up of arginine and zinc sulfate, and the ratio of arginine and zinc sulfate is from 4.62: 1 to 19.24: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 4.62: 1 to 11: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 13.47: 1 to 19.24: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 11: 1 to 19.24: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 6.6: 1 to 19.24: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 9.44: 1 to 19.24: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 4.62: 1 to 13.47: 1.
The ratio of above-mentioned described arginine and zinc sulfate can be from 6.6: 1 to 13.47: 1.
This utility model compared with prior art has following advantage: the quality and quantity that has improved sperm greatly, the activity ratio and the energy of sperm have promptly been improved, reduced rate of teratosperm, side effect is less, arginine medicine and its therapeutic effect of folk prescription zinc sulfate medicine with respect to folk prescription are better, and experimental data is seen experiment 1 in detail.
4, description of drawings:
Below in conjunction with accompanying drawing this utility model is done detailed explanation:
Fig. 1 is the present invention's various dose in toxicity test curve chart corresponding with the laboratory animal mortality rate;
Fig. 2 is that the present invention accurately proofreaies and correct the back various dose curve chart corresponding with probit through FEILLER in toxicity test.
5, the specific embodiment:
A large amount of documents and materials reports, experimental result and clinical proof, zinc is to be present in one of indispensable trace element in the human body.It is low that youth or adult's zinc deficiency will sexual function occur, secondary sex characteristics ateliosis, breast and pubes aplasia.The male can testis occur and weight of prostate alleviates, and sperm quantity reduces, and teratospermia increases, and serum testosterone level obviously descends.Zinc element is the important composition of more than 70 kind of synzyme in the human body, and with the active direct correlation of 200 plurality of enzymes, wide participation male genetic physiological process directly particularly, keep the growth encourage sexual function, improve the quality and quantity of sperm, and participate in the synthetic of testosterone, the activity of can also supplement nutrition spermatogenic epithelium and sperm, and the combination of participant's body protein.
Zinc is very great to the health effect of human body, no matter is baby, child, teenager, adult or old people, all can not zinc deficiency or lose zinc, and especially to the sexual development aspect, more outstanding.And in male sterility, on the problems such as liquefaction of zinc to quantity, vigor, activity ratio, sperm deformity and the seminal fluid of sperm, all play great influence.
Free amino acid participates in the metabolism of sperm in the human refining, is sperm energy supplement source, can promote the activity of sperm.Aminoacid has chelation again, can sperm be shielded in conjunction with deleterious metal ion.Aminoacid also participates in the synthetic of hormone enzyme and other active substances.Influence the synthetic and renewal of above-mentioned substance if aminoacid is not enough, certainly will cause the spermatogenesis obstacle.Studies confirm that recently, arginine has stimulates hypophysis to discharge the effect of promoting sexual gland hormone, oral arginine not only can improve the quantity of sperm, and can improve the quality of sperm, thus can be used as the treatment semen volume not enough and less, the male sterility due to weak, the teratozoospermia.
Preparation method of the present invention is as follows:
1, take by weighing zinc sulfate and starch according to recipe quantity, both grind after mixing, and cross 80 mesh sieves.
2,60 mesh sieves are crossed in arginine, calcium hydrogen phosphate, micropowder silica gel respectively, standby.
3, take by weighing calcium hydrogen phosphate and divide secondary and step (1) obtain the abundant mix homogeneously of material.
4, take by weighing arginine by recipe quantity, and with the abundant mix homogeneously of material that step (3) obtains, add the micropowder silica gel mixing at last, cross 20 mesh sieves.
5, heavily operate at once according to theoretical grain by capsule manufacture technology.
Add the compound recipe that arginine is formed with zinc sulfate, be used for the treatment of the male infertility patient, to improve sperm quantity and sperm quality.After tests such as a series of pharmacology, drug effect, toxicity and clinical practice, prove that curative effect is more satisfactory, and compound recipe is better than folk prescription.Therefore, the capsular prescription of arginine compound has sufficient scientific basis, and being applied to the male sterility treatment has sure curative effect, and toxic and side effects is very little, is a Gospel to the patient, will produce huge economic benefit and social benefit.
Experiment 1:1. arginine, zinc sulfate folk prescription and the former side of smart peace are to the spermatogenesis of normal male mice and CTX model male mice and improve the compound that the former side of the smart ammonia of influence of sperm quality is made up of the zinc sulfate of 220mgD arginine and 20mg.1.1 purpose
The arginine of exploration various dose and zinc sulfate reach the influence to male sexual organ to Spermatogenic action, the raising sperm quality of male mice (mice that comprises the oligospermia model that normal mouse and cyclophosphamide cause) on the former side's of essence peace compound recipe basis, seek the folk prescription optimal dose, provide experimental basis for designing smart peace compound recipe optimum formula.Establish " smart peace capsule " former compound recipe and digital ratio simultaneously, thereby obtain the information whether compound recipe is better than folk prescription.1.2 animal and medicine 1.2.1 laboratory animal
Male NIH kind mice, body weight 18～25g, quality certification 98A030 is provided by Guangdong Province's medical experimental animal center.1.3 test method 1.3.1 experiment contrast group and pathological model
Normal control group: 1% sodium carboxymethyl cellulose group (0.4ml/10g body weight); Positive controls: (make positive control medicine foundation with Methyl testosterone: Methyl testosterone belongs to androgen, can promote that the male genitals grows, and can improve sperm quantity and vigor
[2,14,15], the visible dosage of bibliographical information is that to make the zooperal positive control medicine of Spermatogenic action be successful to 20mg/kg body weight Methyl testosterone
[7]): pathological model: 1.2.2 tests medicine with cyclophosphamide (being called for short CTX)
Arginine (lot number 980701), Kangda Amino-acid Factory of Shanghai's product.Zinc sulfate (lot number 980103-1), the Guangzhou Chemical Reagent Factory product.Smart peace capsule (lot number 980701; Arginase 12 20mg and zinc sulfate 20mg/ capsule; Combination in 11: 1; To call the former side of smart peace in the following text), Zhongshan city three just medical group provides.Methyl testosterone sheet (lot number 980701 first), Shanghai China Xinyi Pharmaceutical Factory product.Face with preceding the medicine grind into powder, be made into required its drug level suspension with 1% sodium carboxymethyl cellulose (preparing with distilled water), it is the 0.4ml/10g body weight that each medicine of above-mentioned filling stomach is irritated gastric capacity mice at every turn.Cyclophosphamide (CTX; Lot number 980607), the product of Hualian Pharmaceutical Co., Ltd., Shanghai.Face the injection that is made into 2mg/ml with water for injection with preceding.Set up pathological model, according to being: discoveries such as Schilky can cause azoospermia, and spermatogenesis reduced during than low dosage when the integral dose of cyclophosphamide reaches 6～10g
[12,13]Attached: the CTX method for establishing model: the result of prerun shows, CTX 20mg/kg ip, and Qd continuous 5 days, can cause mouse sperm to reduce.But after discontinuing medication 21 days, the mouse testis spermatogenesis is existing to be recovered.To compare oligospermia not obvious with normal mouse.For this reason, every day, ipCTX 20mg/kg used 5 days continuously, and later on every 6 days ipCTX 20mg/kg once, successive administration 3 times can successfully be set up mouse sperm and reduce model.
1.3.2 grouping, administration and dissection
180 of male mices, 18～22g divides 18 groups at random.
1.3.2.1 influence to normal male mice
The 1st group～the 9th group is normal mouse, presses the grouping and the dosage ig administration of table 1.1, Qd * 21d, and experiment mice is respectively organized in the dissection in second day after the last administration.
Grouping of table 1.1 normal mouse and dosage
1.3.2.2 influence to the CTX model mice
| Group | Dosage (mg/kg) |
| The 1st group: 1% sodium carboxymethyl cellulose | ????0.4ml/10g |
| The 2nd group: arginine | ????280mg/kg |
| The 3rd group: arginine | ????430mg/kg |
| The 4th group: arginine | ????660mg/kg |
| The 5th group: zinc sulfate | ????46mg/kg |
| The 6th group: zinc sulfate | ????70mg/kg |
| The 7th group: zinc sulfate | ????108mg/kg |
| The 8th group: Methyl testosterone | ????20mg/kg |
| The 9th group: the former side of smart peace | ????240mg/kg |
The 10th group～the 18th group is the model mice (method of setting up model is seen 1.3.1) that CTX causes oligospermia and downgrade, every group of difference ip (CTX) 20mg/kg, Qd * 5d, (each organizes at continuous ipCTX after 5 days, later on once, continuous three times every 6 days ip CTX 20mg/kg.) group and the dosage ig administration that began by table 1.2 in the 6th day of experiment, Qd * 21d after the last administration second day dissects and respectively to organize experiment mice.
Grouping of table 1.2 CTX model mice and dosage
1.3.3 sperm count, sperm motility and motility of sperm and abnormal rate detection method
[13,16～18]:
| Group | Dosage (mg/kg) |
| The 10th group: 1% sodium carboxymethyl cellulose | ????0.4ml/10g |
| The 11st group: arginine | ????280mg/kg |
| The 12nd group: arginine | ????430mg/kg |
| The 13rd group: arginine | ????660mg/kg |
| The 14th group: zinc sulfate | ????46mg/kg |
| The 15th group: zinc sulfate | ????70mg/kg |
| The 16th group: zinc sulfate | ????108mg/kg |
| The 17th group: Methyl testosterone | ????20mg/kg |
| The 18th group: the former side of smart peace | ????240mg/kg |
The male mice execution with experiment in second day after the last administration is got the left side epididymis and is weighed, with reference to people's methods such as Ficsor
[16]Get the left side epididymis in containing the diameter 5cm plate of 2ml phosphate buffer normal saline, shred and make sperm suspension, put in 37 ± 1 ℃ of water-baths and to make it fully free in 15 minutes, then that sperm suspension is one after another drop of on slide, (under 20～25C), high power lens is observed 200 sperm counts down, calculates the sperm motility percentage rate in room temperature.Other gets the 0.1ml seminal fluid and puts into and contain 1.9ml 5% sodium bicarbonate in vitro, shakes up, and counts sperm count on counting chamber.Other gets 1 seminal fluid smear on slide, observes sperm morphology and adds up the sperm deformity percentage rate.1.3.4 data processing method
The weight of animals, sperm count and acropetal coefficient check with t, and sperm motility, motility of sperm, rate of teratosperm are checked with x2.Every index of each group relatively in the experimental result, the computational methods that increase number or reduce the percent of number are that the number that increases or reduce accounts for and is compared several percent.1.4 experimental result
It is all normal that each organizes mice feed, behavior, figure.
The weight of mice of the 1st～9 group (comprising each administration group of matched group and normal mouse) normal (seeing Table 1-A); Hair color is glossy; Testis size normal (testis and epididymis coefficient see Table 1-E); Sperm count, activity ratio, energy and rate of teratosperm (seeing Table 1-C).
The 10th～18 group of (CTX model group) weight of mice situation (seeing Table 1-B), testis size all similar (testis and epididymis coefficient see Table 1-F) to matched group, but hair color lacks gloss, sperm count, activity ratio, energy and rate of teratosperm (seeing Table 1-D).1.4.1 influence to normal group experiment mice sperm and testis and epididymis
(1) the essence former side's group of peace (the 9th group) is compared with matched group (the 1st group), and the mouse sperm number increases by 35.4% (P＜0.05), sperm motility rising 15.0% (P＜0.05), 3 grades of motilities of sperm rising 5.0% (P＞0.05).
(2) mice gives arginase 12 80mg/kg (the 2nd group), 430mg/kg (the 3rd group) and 660mg/kg (the 4th group) respectively, and the mouse sperm number increases by 3.0%, 9.2% and 16.9% than matched group (the 1st group) respectively.But compare with matched group, difference is remarkable (P＞0.05) not.Each organizes sperm motility and 3 grades of motilities of sperm are similar to matched group.Arginine is taken in prompting separately has Spermatogenic action to mice, but its effect is good not as the former side of smart peace.
(3) mice gives zinc sulfate 46mg/kg (the 5th group), 70mg/kg (the 6th group), 108mg/kg (the 7th group) respectively.Compare with matched group (the 1st group), each organizes sperm count increases by 29.2% (P＜0.05), 30.8% (P＜0.05), 32.2% (P＜0.05) respectively.Each organizes sperm motility and 3 grades of motilities of sperm are similar to matched group.Prompting is single with the zinc sulfate generation of dose dependent ground promotion sperm significantly, but its effect is still low than the former general plan of smart peace.
(4) mice gives Methyl testosterone (the 8th group), compares with matched group (the 1st group), and the mouse sperm number increases by 27.7% (P＜0.05), sperm motility rising 10% (P＞0.05), and 3 grades of motilities of sperm are similar to matched group.
(5) give the various dose arginine, zinc sulfate or Methyl testosterone can reduce rate of teratosperm, but because of control group mice abnormal rate very low (1.2 ± 0.6%), deliberately justice is little.
(6) to normal mouse, give the various dose arginine, zinc sulfate and the former side of smart peace, Methyl testosterone all do not have influence (P＞0.05) to mouse testis coefficient and epididymis coefficient.1.4.2 influence to CTX model group damage mouse sperm and testis and epididymis
(1) 1% sodium carboxymethyl cellulose+CTX group (the 10th group) is compared with normal control group (the 1st group), the mouse sperm number reduces by 32.3% (P＜0.05), sperm motility reduces by 30.2% (P＜0.01), 3 grades of sperm motilities reduce by 31.7% (P＜0.01), and rate of teratosperm increases by 66.7% (P＜0.01).
(2) the smart former side group of peace (the 18th group) is compared with CTX group (the 10th group), and sperm count increases by 56.8% (P＜0.01), sperm motility rising 53.5% (P＜0.01), 3 grades of sperm motilities rising 60.0% (P＜0.01).
(3) after mice gives arginase 12 80mg/kg (the 11st group), 430mg/kg (the 12nd group) and 660mg/kg (the 13rd group) respectively, sperm count increases by 36.4% (P＜0.05), 40.9% (P＜0.05), 47.7% (P＜0.05) respectively, sperm motility rising 12.2% (P＞0.05), 15.7% (P＞0.05), 20.5% (P＞0.05), 3 grades of motilities of sperm also increase by 6.4% (P＞0.05), 11.4% (P＞0.05), 29.6% (P＞0.05) respectively.
(4) after mice gives zinc sulfate 46mg/kg (the 14th group), 70mg/kg (the 15th group), 108mg/kg (the 16th group) respectively, sperm count increases by 50.0% (P＜0.05), 52.3% (P＜0.05), 50.0% (P＜0.05) respectively, sperm motility rising 27.2% (P＜0.05), 20.5% (P＞0.05), 15.4% (P＞0.05).
(5) mice gives Methyl testosterone (the 16th group), and sperm count increases by 95.5% (P＜0.01), sperm motility rising 52.3% (P＜0.01).
(6) give various dose arginine, zinc sulfate, the former side of smart peace, Methyl testosterone after, the mouse sperm deformity rate all decreases.Each medication group and CTX model group are the 10th group and compare that rate of teratosperm reduces by 55.0%～80.0% (P＜0.01).
(7) compare two groups of mouse testis coefficient and epididymis coefficients similar (P＞0.05) in mice CTX model group (the 10th～18 group) with matched group (the 1st group).1.5 brief summary
(1) mice respectively ig give arginase 12 80,430 and 660mg/kg and CTX is caused that the oligospermia model mice has more significantly promote the spermatogenesis effect, present dose-effect relationship substantially.Make sperm count increase by 36.4% (P＜0.05), 40.9% (P＜0.05), 47.7% (P＜0.05) respectively.Above-mentioned 3 dosage arginine also have the effect of certain promotion spermatogenesis to normal male mice, but that its effect is nothing like the CTX model mice is good.To normal mouse and CTX model mice Spermatogenic action and raising sperm quality, the effect of arginine 660mg/kg dosage is excellent than other dosage.
(2) after male mice gives zinc sulfate 46,70,108mg/kg respectively, demonstrate Spermatogenic action and improve sperm quality, make the normal mouse sperm count increase by 29.2～32.2%.Making to mouse sperm number behind the CTX increases by 50.0～52.3%, also increases to some extent to normal mouse or to CTX mouse sperm activity ratio.Wherein the heavy dose group is similar than other with improving the sperm quality effect for the Spermatogenic action of zinc sulfate 46mg/kg, and each group difference is remarkable (P＞0.05) not.Therefore, the dosage of zinc sulfate surpasses 46mg/kg, fails further to increase mouse sperm number and motility of sperm.
(3) give smart peace former side 240mg/kg, can make the sperm count of normal mouse and CTX model mice increase by 35.4% (P＜0.05) and 56.8% (P＜0.01) respectively, sperm motility also increases by 15.0% (P＜0.05) and 53.5% (P＜0.01) respectively, and 3 grades of motilities of sperm increase by 5.0% (P＞0.05) and 60.0% (P＜0.01) respectively.It is more excellent that the effect of arginine (280,430,660mg/kg) or zinc sulfate (46,70,108mg/kg) is used in this effect more separately, but optimum efficiency not necessarily.
(4) give Methyl testosterone 20mg/kg body weight after, normal male mice and CTX model mice all there are obvious Spermatogenic action and improve sperm quality, sperm count increases by 27.7% (P＜0.05) and 95.5% (P＜0.01) respectively, sperm motility raise respectively 10.0% (P＞0.05) and 52.3% (P＜0.01).
(5) give arginine, zinc sulfate, the former side of smart peace, Methyl testosterone after, all can reduce CTX and produce mouse sperm deformity rate (P＜0.01).
(6) the former side of smart peace, Methyl testosterone do not have influence to the weight coefficient of testis and the epididymis coefficient of male mice.
This result of study shows that aspect spermatogenesis and raising sperm quality, arginine and zinc sulfate folk prescription have certain effect to male mice, and be more remarkable to CTX model mice spermatogenesis and raising sperm quality.The best effective dose of arginine is 660mg/kg, and the best effective dose of zinc sulfate is 46mg/kg.Arginine or zinc sulfate that various dose is used in the smart peace former side's of compound recipe (combination of arginine and zinc sulfate) Spermatogenic action and the effect of raising sperm quality more separately are strong.Arginine and zinc sulfate folk prescription, the smart former side of peace and Methyl testosterone all do not have influence to the testis and the epididymis coefficient of male mice.
Table 1-A ig arginine, zinc sulfate and the former side of smart peace are to the influence (x ± SD of normal group experiment mice body weight; N=10)
0 1 2 3 41. 1％ 22.3±1.2 31.4±2.4 33.1±2.9 34.3±2.2 35.6±2.32. 280mg／kg 22.2±1.3 31.1±2.2 33.2±2.3 33.9±2.4 34.6±2.4*3. 430mg／kg 21.7±1.6 28.9±1.6 31.1±1.8 34.0±1.9 35.2±2.2*4. 660mg／kg 21.9±1.1 31.1±1.4 31.9±2.5 32.6±3.2 33.7±3.2*5. 46mg／kg 22.2±1.9 29.5±2.2 32.6±2.5 34.0±2.5 33.9±3.3*6. 70mg／kg 22.5±2.1 28.3±3.1 30.9±3.1 33.9±3.1 35.3±1.5*7. 108mg／kg 22.8±1.3 32.2±2.1 34.4±2.7 35.0±2.4 34.7±2.7*8. 20mg／kg 22.1±1.2 32.7±1.5 34.2±1.2 36.0±2.1 36.8±2.7*9. 240mg／kg 22.2±1.2 30.7±2.2 32.8±3.7 31.1±3.5 34.4±3.4**P＞0.05,1-B ig ； Zinc sulfate and the former side of smart peace are on the CD (impact (x ± SD of model group damage Mouse Weight; N=10)
0 1 2 3 410.CTX20mg／kg＋1％ 22.1±1.7 28.7±2.6 30.6±4.6 33.7±3.7 35.2±3.6*11.CTX20mg／kg＋280mg／kg 22.0＝1.2 27.7±1.2 30.2±2.7 32.3±2.4 33.2±2.9*12.CTX20mg／kg-430mg／kg 21.2＝1.0 29.1±2.4 30.5±3.0 32.9±3.0 34.0±3.3*13.CTX20mg／kg＋660g／kg 21.8±1.6 28.6±1.1 21.4±1.4 33.0±1.8 34.5±2.7*14.CTX20mg／kg＋46mg／kg 21.9±1.3 29.0±1.8 31.0±1.8 32.1±3.6 34.0±2.9*15.CTX20mg／kg＋70mg／kg 22.0±1.5 28.7±2.5 31.0±2.3 31.0±2.3 32.9±1.9*16.CTX20mg／kg＋108mg／kg 21.7±1.1 29.4±1.3 33.1±1.4 34.8±1.7 34.6±2.2*17.CTX20mg／kg＋20mg／kg 21.6±2.2 29.0±2.3 31.1±2.1 33.7±2.5 34.2±2.3*18.CTX20mg／kg＋240mg／kg 21.9±1.2 29.3±2.1 31.9±3.2 32.6±2.3 32.6±3.2**P＞0.05,1-11-C ig； Zinc sulfate and the former side of smart peace are on the impact (x ± SD of normal group experiment mice sperm; N=10)
Sperm count activity ratio motility of sperm (%) abnormal rate group
(* 10
6/ml ) ( ％ ) 3 ( ％ ) 2 ( ％ ) 1 ( ％ ) ( ％ ) 1 6.5±2.3 62.2±10.3 32.2±5.1 21.1±6.6 8.9±5.0 1.2±0.62 6.7±1.9 * 60.3±7.6 * 31.7±4.7 * 18.8±4.3 * 9.8±4.4 * 1.1±1.0 *3 7.1±1.9 * 63.4±12.2 * 32.5±6.1 * 20.3±9.9 * 1.0±7.0 * 1.6±3.7 *4 7.6±1.8 * 64.7±11.3 * 30.6±4.2 * 22.2±7.3 * 11.9±6.3 * 0.6±0.5 **5 8.4±1.6 ** 61.5±4.8 * 33.2±3.8 * 18.8±2.9 * 9.5±4.0 * 0.6±0.7 **6 8.5±2.3 * 62.0±11.3 * 33.1±5.2 * 19.3±6.9 * 9.6±4.8 * 0.7±1.2 **7 8.6±2.2 * 63.0±16.8 * 32.4±4.8 * 20.0±10.9 * 10.6±8.0 * 0.9±0.5 **8 8.3±1.2 ** 68.4±12.6 * 32.4±5.1 * 23.2±8.7 * 12.8±6.3 * 0.6±0.6 **9 8.8±2.0 * 71.5±9.1 * 33.7±4.4 * 24.9±4.0 * 12.9±5.2 * 1.5±0.6 *1,*：P＞0.05； *: P＜0.05; 1 group:1% sodium carboxymethyl cellulose; 2 groups:arginase, 12 80mg/kg; 3 groups:arginine 430mg/kg; 4 groups:arginine 660mg/kg; 5 groups:zinc sulfate 46mg/kg; 6 groups:zinc sulfate 70mg/kg; 7 groups:zinc sulfate 108mg/kg; 8 groups:methyltestosterone 20mg/kg:9 group:the former side 240mg/kg table of smart peace 1-D ig arginine, and and and and and zinc sulfate and the former side of smart peace are to the influence of CTX model group damage mouse sperm (sperm count activity ratio motility of sperm (%) the abnormal rate group of x ± SD:n=10)
(* 10
6/ml ) ( ％ ) 3 ( ％ ) 2 ( ％ ) 1 ( ％ ) ( ％ ) 10 4.4±1.7 43.4±11.9 22.0±6.7 14.9±5.5 6.5±3.4 2.0±0.611 6.0±1.4 * 48.7±10.7 * 23.4±6.9 * 16.0±3.6 * 9.3±4.2 * 0.8±0.6 ***12 6.2±1.9 * 50.2±9.7 * 24.5±7.6 * 17.8±1.8 * 7.9±3.4 * 0.8±0.5 ***13 6.5±1.6 * 52.3±11.6 * 28.5±7.0 * 15.4±6.6 * 8.4±5.8 * 0.6±0.6 ***14 6.6±1.6 ** 55.2±9.8 * 28.6±6.3 * 17.1±6.5 * 9.5±3.3 * 0.9±1.0 ***15 6.7±2.3 * 52.3±10.7 * 27.8±5.2 * 16.4±6.1 * 8.1±4.0 * 0.5±0.6 ***16 6.6±2.0 * 50.1±12.3 * 27.8±6.3 * 14.1±6.0 * 8.2±3.8 * 0.7±0.7 ***17 8.6±1.3 *** 66.1±10.4 *** 32.5±2.7 ** 21.8±3.3 *** 11.9±6.7 ** 0.4±0.4 ***18 6.9±2.0 ** 66.6±11.1 *** 35.2±5.6 ** 20.5±5.1 ** 10.9±4.4 ** 0.6±0.8 ***1,*：P＞0.05； *: P＜0.05; *: P＜0.0110 group:CTX 20mg/kg+1% sodium carboxymethyl cellulose:11 groups:CTX 20mg/kg+ arginase, 12 80mg/kg:12 group:CTX 20mg/kg+ arginine 430mg/kg:13 group:CTX 20mg/kg+ arginine 660mg/kg:14 group:CTX 20mg/kg+ zinc sulfate 46mg/kg; 15 groups:CTX 20mg/kg+ zinc sulfate 70mg/kg:16 group:CTX 20mg/kg+ zinc sulfate 108mg/kg; 17 groups:CTX 20mg/kg+ Methyl testosterone 20mg/kg:18 group:the smart peace of CTX 20mg/kg+ compound recipe 240mg/kg table 1-E ig arginine, zinc sulfate and the former side of smart peace are to the influence (x ± SD of normal group experiment mice testis and epididymis coefficient; N=10)
Table l-F ig arginine is compared in the smart peace of group weight coefficient of testis epididymis coefficient 1.1% sodium carboxymethylcellulose 0.00273 ± 0.00038 0.00098 ± 0.000232. arginase 12 80mg/kg, 0.00279 ± 0.00040*, 0.00095 ± 0.00017*3. arginine 430mg/kg, 0.00263 ± 0.00037*, 0.00088 ± 0.00014*4. arginine 660mg/kg, 0.00291 ± 0.00048*, 0.00103 ± 0.00019*5. zinc sulfate 46mg/kg, 0.00279 ± 0.00036*, 0.00097 ± 0.00010*6. zinc sulfate 70mg/kg, 0.00262 ± 0.00029*, 0.00089 ± 0.00011*7. zinc sulfate 108mg/kg, 0.00283 ± 0.00028*, 0.00096 ± 0.00012*8. first a kind of sedge stosterone 20mg/kg, 0.00275 ± 0.000378*, 0.00092 ± 0.00009*9. former side 240mg/kg 0.00273 ± 0.00038* 0.00094 ± 0.00007**P＞0.05 with Normal group, zinc sulfate and the former side of smart peace are on the impact (x ± SD of CTX model group damage mouse testis and attached highland coefficient; N=10)
lO.CTX20mg/kg+1％ 0.00292±0.00064* 0.00097±0.00016*11.CTX20mg/kg+280mg/kg 0.00279±0.00035* 0.00101±0.00011*12.CTX20mg/kg+430mg/kg 0.00270±0.00031* 0.00096±0.00013*13.CTX20mg/kg+660mg/kg 0.00273±0.00078* 0.00098±0.00011*14.CTX20mg/kg+46mg/kg 0.00281±0.00028* 0.00097±0.00022*15.CTX20mg/kg+70mg/kg 0.00273±0.00044* 0.00101±0.00015*16.CTX20mg/kg+108mg/kg 0.00266±0.00025* 0.00092±0.00013*17.CTX20mg/kg+20mg/kg 0.00266±0.00031* 0.00104±0.00017*18.CTX20mg/kg+240mg/kg 0.00273±0.00057* 0.00101±0.00017**P＞0.05,1-52：2．CTX2.1
Basis based on the folk prescription experiment, cause in the male mice of oligospermia and downgrade model normal male mice and CTX, by fixing arginic dosage and change the dosage of zinc sulfate and fixedly zinc sulfate dosage and change arginic dosage, filter out the best proportioning of this two medicines Spermatogenic action and raising sperm quality, to determine Spermatogenic action and to improve the strongest new smart compound recipe prescription of pacifying of sperm quality.2.2 animal and medicine 2.2.1 laboratory animal
Male NIH kind mice, body weight 18～25g, quality certification 98A030 is provided by Guangdong Province's medical experimental animal center.2.2.2 experiment medicine
Arginine (lot number 980701), Kangda Amino-acid Factory of Shanghai's product.Zinc sulfate (lot number 980103-1), the Guangzhou Chemical Reagent Factory product.Smart peace capsule (lot number 980701; Arginase 12 20mg and zinc sulfate 20mg/ capsule; Combination in 11: 1; To call the former side of smart peace in the following text), Zhongshan city three just medical group provides.Methyl testosterone sheet (lot number 980701 first), Shanghai China Xinyi Pharmaceutical Factory product.Face with preceding the medicine grind into powder, be made into required its drug level suspension with 1% sodium carboxymethyl cellulose (preparing with distilled water), it is the 0.4ml/10g body weight that each medicine of above-mentioned filling stomach is irritated gastric capacity mice at every turn.Cyclophosphamide (CTX; Lot number 980607), the product of Hualian Pharmaceutical Co., Ltd., Shanghai.Face the injection that is made into 2 mg/ml with water for injection with preceding.2.3 the foundation of the smart peace of experimental technique 2.3.1 design dosage
The prompting of folk prescription experimental result, respectively ig arginine and the zinc sulfate effect that can under this experiment condition, obtain Spermatogenic action in various degree and improve sperm quality at normal male mice and CTX model mice, but the Spermatogenic action of folk prescription and improve sperm quality and all be weaker than the smart former side of peace.This experiment intend to adopt fixedly arginine dosage (660mg/kg) change zinc sulfate dosage or fixedly zinc sulfate dosage (49mg/kg) change arginine dosage and form a plurality of compound recipes, filter out Spermatogenic action and the strongest best of breed of raising sperm quality.2.3.2 the essence that the arginine of various dose and zinc sulfate are formed peace compound recipe
Based on the basis of folk prescription experiment, arginine and zinc sulfate are combined into 5 smart peace compound dose groups (group is apart from 0.7) mutually, and each is organized dosage and sees Table 2-1, establish the former side's group of smart peace simultaneously and participate in the prescription screening.
The essence peace compound recipe that the arginine of table 2-1 various dose and zinc sulfate are formed
2.3.3 experiment contrast group and model
| Group | Arginine (mg/kg) | Zinc sulfate (mg/kg) | Ratio |
| 5 groups of 4 groups of smart peace compound recipes of 3 groups of smart peace compound recipes of 2 groups of smart peace compound recipes of 1 group of smart peace compound recipe of the smart peace of the former side's group of smart peace compound recipe | ????220.0 ????660.0 ????462.0 ????323.4 ????660.0 ????660.0 | ????20.0 ????70.0 ????70.0 ????70.0 ????49.0 ????34.3 | ??11∶1 ??9.43∶1 ??6.6∶1 ??4.62∶1 ??13.47∶1 ??19.24∶1 |
Normal control group: 1% sodium carboxymethyl cellulose group (0.4ml/10g); Positive controls: Methyl testosterone group (20mg/kg body weight); Pathological model: set up pathological model with CTX.Attached: the male mice oligospermia that CTX causes and the foundation of downgrade pathological model (setting up model) 2.3.4 grouping, administration and dissection by 1.3.1 joint method
192 of NIH kind male mices, 20～25g divides equally 16 groups at random, and 12 every group, each is organized body weight and distributes roughly close.The 1st group～the 8th group is normal mouse, and the 9th～16 group is CTX model mice group, sees Table 2-2.
The 1st group～the 8th group by table 2-1 dosed administration, every day ig once, continuous 21 days.Set up model by 2.3.3 joint method for the 9th group～the 16th group, each organizes ip CTX 20mg/kg, Qd, continuous 5 days, the 6th day of experiment began the dosed administration by table 2-1, every day ig once, continuous 21 days.Every group of each gastric infusion capacity is the 0.4ml/10g body weight.
After the last administration second day put to death and respectively organized mice, takes out mice left testes and epididymis and weighs, and calculates its organ coefficient, and and takes out the left side epididymis and detect each Mus sperm count, sperm motility and energy and teratospermia number.2.3.5 sperm count, sperm motility and energy and teratospermia are counted detection method and are pressed 1.3.3 joint method.
The grouping of table 2-2 normal mouse and CTX model mice
2.3.6 data processing method
| Group number | Normal mouse | Group number | The CTX model mice |
| ????1 ????2 ????3 ????4 ????5 ????6 ????7 ????8 | 5 groups of Methyltestosterone groups of 1 group of smart peace compound of the smart peace of the 1% carboxymethyl cellulose group former side of smart peace compound, 2 groups of smart peace compounds, 4 groups of smart peace compounds of 3 groups of smart peace compounds | ????9 ????10 ????11 ????12 ????13 ????14 ????15 ????16 | 5 groups of Methyltestosterone groups of 1 group of smart peace compound of the smart peace of the 1% carboxymethyl cellulose group former side of smart peace compound, 2 groups of smart peace compounds, 4 groups of smart peace compounds of 3 groups of smart peace compounds |
The weight of animals, sperm count and acropetal coefficient check with t, sperm motility, motility of sperm, rate of teratosperm x
2Check.Every index of each group relatively in the experimental result, the computational methods that increase number or reduce the percent of number are that the number that increases or reduce accounts for and is compared several percent.2.4 experimental result
It is all normal that each organizes experiment mice feed, behavior, figure.
The mice body weight (seeing Table 2-3) of the 1st group～the 8th group (comprising normal control group and each administration group) increases normal; Hair color is glossy; Testis size normal (testis and epididymis coefficient see Table 2-7); Sperm count, activity ratio, energy and rate of teratosperm see Table 2-5.The 9th group～mice body weight (seeing Table 2-4) of the 16th group (CTX model group) increases normal; Hair color lacks gloss; Testis size normal (testis and epididymis coefficient see Table 2-8); Sperm count, activity ratio, energy and rate of teratosperm see Table 2-6.2.4.1 influence to normal group experiment mice sperm
(1) the 1st group of mice is only given 1% sodium carboxymethyl cellulose, as normal control.Sperm count is 7.4 ± 1.0 * 10
6/ ml, sperm motility are 58.0 ± 5.7%, and sperm deformity rate is 0.6 ± 0.3%.
(2) the 2nd groups of (the former side 240mg/kg of smart peace) mices are recommended the former side of smart peace to the pharmaceutical factory, compare with matched group, sperm count increases by 32.4% (P＞0.05) sperm motility and increases by 13.6% (P＞0.05), and the smart peace of prompting former side have Spermatogenic action and improve sperm quality normal mouse.
(3) the 3rd～7 groups (3 groups: arginine 660mg/kg zinc sulfate 70mg/kg; 4 groups: arginine 462mg/kg zinc sulfate 70mg/kg; 5 groups. arginine 305.4mg/kg zinc sulfate 70mg/kg; 6 groups: arginine 660mg/kg zinc sulfate 49mg/kg; 7 groups: arginine 560mg/kg zinc sulfate 34.3mg/kg) the mice arginine that gives the various dose proportioning respectively adds zinc sulfate.Compare with matched group, each organizes the mouse sperm number increases by 18.9～40.5%, and sperm motility raises 1.0～18.3%.The sperm count and the sperm motility of wherein the 6th group (giving arginine 660mg/kg and zinc sulfate 49mg/kg) increase at most, increase by 40.5% (P＜0.05) and 18.3% (P＞0.05) more respectively with matched group, be higher than the former side's group of smart peace, other groups (3,4,5, the 7 groups) arginine that mice gave or the dosage of zinc sulfate all are different from the 6th group, but their spermatogenesis effect all be not so good as the 6th group good.The sperm count of the 6th group of mice is than Methyl testosterone group slightly high (P＞0.05).This shows that to normal mouse, smart peace prescription is equipped with zinc sulfate 49mg/kg with arginine 660mg/kg, promote that spermatogenesis and the effect of raising sperm quality are the strongest, this is the best spermatogenesis prescription of smart peace.
(4) the 8th groups of (Methyl testosterone 20mg/kg) mices are given Methyl testosterone, compare with matched group, sperm count increases by 31.1% (P＜0.01), and sperm motility increases by 13.5% (P＞0.05), and the prompting Methyl testosterone can promote the generation of normal mouse sperm and improve sperm quality.
(5) give the mice abnormal rate of different formulations smart peace compound recipe or Methyl testosterone similar to matched group.2.4.2 influence to CTX model group damage mouse sperm
Irritate and give 1% sodium carboxymethyl cellulose (1) the 9th group of (CTX model) every day.Compare with matched group, this group mouse sperm number reduces by 35.1% (P＜0.05), sperm motility reduces by 32.1% (P＜0.05), and rate of teratosperm increases by 866.7% (P＜0.01), and prompting is made the downtrod model success of male mice testicular spermatogenic function with CTX.
(2) the 10th groups of (CTX models; The former side 240mg/kg of smart peace) the former side of the smart peace of ig, compare with the 9th group (CTX model), sperm count increases by 70.8% (P＜0.01), sperm motility increases by 40.9% (P＜0.05), and rate of teratosperm reduces the effect that 24.1% (P＜0.05) prompting has tangible promotion spermatogenesis and improves sperm quality.After suppressing mouse testis with CTX, former its functional rehabilitation that can make of smart peace is normal.
(3) the 11st～15 groups of (CTX models; 11 groups: CTX 20mg/kg arginine 660mg/kg+ zinc sulfate 70mg/kg; 12 groups: CTX 20mg/kg+ arginine 462mg/kg+ zinc sulfate 70mg/kg; 13 groups: CTX 20mg/kg+ arginine 305.4mg/kg+ zinc sulfate 70mg/kg; 14 groups: CTX 20mg/kg+ arginine 660mg/kg+ acid zinc 49mg/kg; 15 groups: CTX 20mg/kg+ arginine 660mg/kg+ zinc sulfate 34.3mg/kg; ) mice, give the arginine and the zinc sulfate of various dose proportioning respectively.Compare with the 9th group, each organizes the mouse sperm number increases by 31.3～104.2%, sperm motility raises 48.5～69.0%, 3 grades of motilities of sperm increase by 44.2%～62.2% (P＜0.01), rate of teratosperm reduces by 25.9～34.5% (P＜0.05), the sperm count and the sperm motility of wherein the 14th group (giving arginine 660mg/kg and zinc sulfate 49mg/kg) increase at most, increase by 104.2% (P＜0.01) and 69.0% (P＜0.01) respectively, the sperm count of the former side's group of smart peace and sperm motility be high by 19.5% (P＜0.05) and 20.0% (P＜0.01) respectively.Wherein the increase of sperm count in addition than normal control group also high by 32.4% (P＜0.05) than Methyl testosterone group also high by 22.5% (P＜0.05).Other groups (11,12,13, the 15 groups) arginine that mice gave or the dosage of zinc sulfate are different from the 14th group, but their spermatogenesis effect all be not so good as the 14th group good.The reduction (34.5%) of the 14th group of mouse sperm deformity rate is also more remarkable than other groups.This shows that (arginine: Spermatogenic action zinc sulfate=13.47: 1) and raising sperm quality are the strongest, and it is best to reduce the rate of teratosperm effect, and this is the best spermatogenesis prescription of smart peace to be equipped with zinc sulfate 49mg/kg with arginine 660mg/kg in the smart peace prescription.
(4) the 16th groups of (CTX 20mg/kg+ Methyl testosterone 20mg/kg) mices are compared with the 9th group, sperm count increases by 66.7% (P＜0.01), sperm motility increases by 51.8% (P＜0.05), after rate of teratosperm reduced by 25.9% (P＜0.05) prompting CTX inhibition mouse testis, Methyl testosterone just its spermatogenesis recovered normal.2.4.3 influence to normal group and CTX model group experiment mice testis and epididymis
Give essence peace compound recipe or Methyl testosterone that normal mouse and CTX model mice give various prescriptions respectively, each organizes mouse testis and the epididymis coefficient is compared with matched group and indifference (P＞0.05) (seeing Table 2-7 and table 2-8).2.5 brief summary
(1) gives the arginine of various dose proportioning to normal mouse and zinc sulfate is irritated stomach, can increase sperm count 18.9～40.5%.Wherein arginine 660mg/kg adds zinc sulfate 49mg/kg group and makes the mouse sperm number increase by 40.5% (P＜0.05).(arginine: essence peace compound recipe zinc sulfate=13.47: 1) can promote the generation of mouse sperm to point out this prescription.Though other groups also make the mouse sperm number increase to some extent, with matched group difference not significantly (P＞0.05).The smart peace of various prescriptions compound recipe does not have obvious influence (P＞0.05) to normal mouse sperm activity ratio and abnormal rate.
(2) essence of the arginine of various dose proportioning and zinc sulfate peace compound recipe all can recover owing to giving the mouse testis spermatogenesis that CTX suppresses.Wherein arginine 660mg/kg adds the prescription best results of zinc sulfate 49mg/kg.With only compare to CTX group (the 9th group), it makes sperm count increase by 104.2% (P＜0.01), normal matched group high by 32.4% (P＜0.05) is than Methyl testosterone group high by 22.5% (P＜0.05).This group sperm motility also increases by 69.0% (P＜0.01).The Spermatogenic action of other groups and raising sperm quality are not as this group.It is the best prescription that impels CTX oligospermia model mice Spermatogenic action and improve sperm quality that this group rate of teratosperm also reduce at most (34.5%, P＜0.05) prompting arginine 660mg/kg to add zinc sulfate 49mg/kg.
(3) Methyl testosterone 20mg/kg/ angel normal mouse sperm count increases by 31.1% (P＜0.01), sperm motility rising 13.5% (P＞0.05).Methyl testosterone also can recover owing to giving the mouse testis spermatogenesis that CTX suppresses, with only compare to CTX group, it makes sperm count increase by 66.7% (P＜0.01), and sperm motility increases by 51.8% (P＜0.05), and rate of teratosperm also reduces by 25.9% (P＜0.05).
(4) the different essence of various dosage ratios peace compound recipe and Methyl testosterone all do not have influence to the testis and the epididymis coefficient of normal mouse and CTX model mice.
(5) smart each compound recipe of peace does not have influence to mice body weight and genitals's coefficient.
This result of study shows, the male mice of the oligospermia model of making to normal male mice and of CTX, by fixing arginic dosage and change zinc sulfate dosage and fixedly zinc sulfate dosage and change arginic dosage, determine that butyrin peace compound recipe Spermatogenic action and the strongest novel composing prescription of raising sperm quality are: arginine 660mg/kg and zinc sulfate 49mg/kg.The optimum dose proportion of two medicine Spermatogenic actions and raising sperm quality is 13.47: 1.This novel composing prescription is better than the former side of smart peace.The arginine of table 2-3.ig various dose proportioning, zinc sulfate are to the influence (x ± SD of normal group mice body weight (g); N=12)
0 1 2 3 41. 27.8±2.2 29.7±2.4 30.9±2.1 30.4±3.0 30.4±3.42. ( 240mg/kg ) 28.4±1.3 30.2±1.3 30.5±0.9 31.0±1.4 30.9±2.3*3.660mg/kg70mg/kg 27.6±2.3 31.1±2.1 32.3±1.9 33.6±2.7 33.8±3.5*4.462mg/kg70mg/kg 28.0±1.8 31.1±1.8 31.4±1.8 32.3±2.4 32.7±2.9*5.305.4mg/kg70mg/kg 27.3±2.0 29.2±3.7 29.4±4.2 28.7±4.6 27.8±4.4*6.660mg/kg49mg/kg 27.6±2.8 29.6±2.6 29.9±4.1 31.6±4.5 32.3±3.6*7.660mg/kg34.3mg/kg 26.5±2.4 29.1±2.3 29.7±3.2 31.2±4.1 32.2±4.0*8.20mg/kg 27.9±1.6 30.7±2.0 31.5±2.4 33.6±2.4 34.0±2.6**P＞0.05,
The arginine of table 2-4. ig various dose proportioning, zinc sulfate are to the influence (x ± SD of CTX model group mice body weight (g); N=12)
0 1 2 3 49.CTX20mg/kg 27.2±2.3 27.8±2.9 28.6±4.0 31.0±4.7 29.3±3.6*10.CTX20mg/kg240mg/kg 27.1±1.9 29.0±2.2 29.8±2.2 30.9±2.9 30.2±2.8*11.CTX20mg/kg660mg/kg70mg/kg 27.8±1.6 28.9±2.5 29.1±3.1 31.6±3.2 31.8±2.9*12.CTX20mg/kg462mg/kg70mg/kg 26.8±2.4 26.4±2.3 25.7±3.3 27.4±3.9 28.9±3.1*13.CTX20mg/kg305.4mg/kg70mg/kg 27.4±1.1 28.3±1.9 28.5±1.6 30.7±2.0 30.3±2.5*14 CTX20mg/kg660mg/kg49mg/kg 27.5±1.2 27.1±1.3 29.4±1.6 28.9±2.5 27.6±2.4*15.CTX20mg/kg660mg/kg34.3mg/kg 27.0±1.8 26.2±1.4 27.7±1.3 29.0±2.0 29.2±2.8*16.CTX20mg/kg20mg/kg 28.0±2.3 28.6±2.0 30.8±2.7 32.7±2.4 32.6±3.3**P＞0.05,
The arginine of table 2-5. ig various dose proportioning, zinc sulfate are to the influence (x ± SD of normal group mouse sperm; N=12) group motility of sperm
Sperm count (* 10
6/ ml) activity ratio (%) rate of teratosperm (%) is other
3 2 11 7.4±1.0 58.0±5.7 33.8±8.8 17.4±5.5 6.9±4.0 0.6±0.32 9.8±1.3 ** 65.9±5.4 * 36.9±5.9 * 20.1±4.3 8.9±2.8 0.6±0.2 *3 9.6±1.3 ** 58.4±11.8 * 31.8±6.5 * 18.6±3.8 8.0±3.7 0.6±0.1 *4 9.8±1.8 ** 61.4±9.1 * 33.0±9.4 * 21.5±5.7 6.9±3.2 1.0±1.3 *5 8.8±1.7 * 62.5±6.9 * 34.4±5.3 * 19.8±2.8 8.3±2.3 0.8±0.5 *6 10.4±2.6*** 68.6±9.1 *** 37.4±5.9 ** 21.4±3.6 9.8±3.7 0.5±0.1 *7 9.6±1.6 * 59.4±10.8 * 32.3±6.9 * 18.1±3.2 9.1±2.7 0.6±0.2 *8 9.7±1.8 *** 65.8±6.9 ** 37.9±5.2 ** 20.0±3.1 7.9±3.0 0.7±0.2 *1，*：P＞0.05：**：P＜0.05； *: P＜0.01:1 group:1% sodium carboxymethyl cellulose 0.5ml/10g; 2 groups:smart peace former side (240mg/kg); 3 groups:arginine 660mg/kg zinc sulfate 70mg/kg; 4 groups:arginine 462mg/kg zinc sulfate 70mg/kg; 5 groups. arginine 305.4mg/kg zinc sulfate 70mg/kg; 6 groups:arginine 660mg/kg zinc sulfate 49mg/kg; 7 groups:arginine 660mg/kg zinc sulfate 34.3mg/kg; 8 groups:Methyl testosterone 20mg/kg.
The arginine of table 2-6. ig various dose proportioning, zinc sulfate are to the influence (x ± SD of CTX model group mouse sperm; N=12) the other sperm count (* 10 of group motility of sperm
6/ ml) activity ratio (%)
3 grades of 2 grades 1 grade rate of teratosperms (%) 9 4.8 ± 2.3 39.4 ± 8.3 21.7 ± 5.3 12.1 ± 4.0 5.7 ± 3.2 5.8 ± 1.710 8.2 ± 2.0
* *55.5 ± 7.8
*29.3 ± 5.9
*16.9 ± 3.2 9.0 ± 4.0 4.4 ± 1.2
*11 7.3 ± 1.2
* *62.6 ± 6.8
*32.0 ± 4.7
*20.5 ± 3.0 10.1 ± 3.0 4.0 ± 1.7
*12 7.4 ± 2.2
*63.5 ± 7.1
*33.5 ± 5.7
*18.8 ± 2.9 11.5 ± 3.5 4.2 ± 1.9
*13 6.3 ± 1.3
*62.0 ± 7.8
*35.2 ± 3.0
*19.5 ± 4.3 9.0 ± 3.1 4.3 ± 1.7
*14 9.8 ± 1.5
* *66.6 ± 7.2
* *5.2 ± 3.0
*21.0 ± 3.0 10.4 ± 4.5 3.8 ± 1.7
*15 8.5 ± 2.9
* *58.5 ± 9.5
*31.3 ± 6.7
*18.2 ± 4.4 9.0 ± 3.9 4.1 ± 2.1
*16 8.0 ± 2.2
* *59.8 ± 5.0
*33.8 ± 5.4
*19.7 ± 3.5 6.3 ± 3.8 4.3 ± 1.1
*With 1 group of comparison, *: P＞0.05; *: P＜0.05; * *: P＜0.01; With 9 groups of comparisons, *: P＞0.05; *: P＜0.05; * *: P＜0.019 group: CTX 20mg/kg+1% sodium carboxymethyl cellulose; 10 groups: the former side 240mg/kg of the smart peace of CTX 20mg/kg+; 11 groups: CTX 20mg/kg arginine 660mg/kg zinc sulfate 70mg/kg; 12 groups: CTX 20mg/kg arginine 462mg/kg zinc sulfate 70mg/kg; 13 groups: CTX 20mg/kg arginine 305.4mg/kg zinc sulfate 70mg/kg; 14 groups: CTX 20mg/kg arginine 660mg/kg zinc sulfate 49mg/kg; 15 groups: CTX 20mg/kg arginine 660mg/kg zinc sulfate 34.3mg/kg; 16 groups: CTX20mg/kg Methyl testosterone 20mg/kg
The arginine of table 2-7 ig various dose proportioning and zinc sulfate are to the influence (x ± SD of normal group mouse testis, epididymis coefficient; N=12)
The smart peace of group weight coefficient of testis epididymis coefficient 1.1% sodium carboxymethylcellulose 0.00313 ± 0.00052 0.00111 ± 0.000212. former side (240mg/kg) 0.00302 ± 0.00028* 0.00099 ± 0.00011*3. arginine 660mg/kg zinc sulfate 70mg/kg 0.00299 ± 0.00044* 0.00093 ± 0.00014*4. arginine 462mg/kg zinc sulfate 70mg/kg 0.00306 ± 0.00039* 0.00102 ± 0.00012*5. arginine 305.4mg/kg zinc sulfate 70mg/kg 0.00324 ± 0.00065* 0.00100 ± 0.00019*6. arginine 660mg/kg zinc sulfate 49mg/kg 0.00296 ± 0.00038* 0.00102 ± 0.00012*7. arginine 660mg/kg zinc sulfate 34.3mg/kg 0.00290 ± 0.00026* 0.00101 ± 0.00014*8. Methyltestosterone 20mg/kg 0.00293 ± 0.00034* 0.00102 ± 0.00008**P＞0.05 is compared with Normal group
The arginine of table 2-8ig various dose proportioning and zinc sulfate are to the influence (x ± SD of CTX model group mouse testis, epididymis coefficient; N=12)
9.CTX20mg/kg1％ 0.00298±0.00049* 0.00110±0.00020*10.CTX20mg/kg240mg/kg 0.00287±0.00026* 0.00100±0.00017*11.CTX20mg/kg660mg/kg70mg/kg 0.00277±0.00029* 0.00101±0.00013*12.CTX20mg/kg462mg/kg70mg/kg 0.0031±0.00035* 0.00116±0.00022*13CTX20mg/kg305.4mg/kg70mg/kg 0.00272±0.00024* 0.00106±0.00011*14CTX20mg/kg660mg/kg49mg/kg 0.00319±0.00050* 0.00112±0.00015*15.CTX20mg/kg660mg/kg34.3mg/kg 0.00308±0.00063* 0.00106±0.00016*16.CTX20mg/kg20mg/kg———— 0.00279±0.00030* 0.00099±0.00010**P＞0.05,3：
14.1 summary: smart peace capsule is by arginine and zinc sulfate (ZnSO
2.7H
2O) forming, is white powder, water-soluble.This experiment is done acute toxicity test with the NIH its mouse oral, as a result its LD
50Be 6268.9mg/kg, the 95% credible 5758.2～6825.0mg/kg that is limited to.
14.2 test objective: observe NIH its mouse oral filling stomach and give arginine and zinc sulfate (ZnSO
2.7H
2O) producing property of raw material powder toxic reaction and death condition are for further toxicity test provides foundation.
14.3 tried thing:
14.3.1 title: smart peace capsule raw material powder is by arginine and zinc sulfate (ZnSO
2.7H
2O) by be made at 13.47: 1 by Zhongshan city three just medical group company provide.Zinc sulfate: lot number: 990601 pharmaceutical grade, the Taishan, Guangdong Xinning pharmaceutical factory product arginine: lot number: 991011, pharmaceutical grade, Kangda Amino-acid Factory of Shanghai produces
14.4 animal:
14.4.1 the source, kind: the NIH mice that provides by Guangdong Medical Lab Animal Center (quality certification number: 99A030)
14.4.2 body weight: 18～22 grams
14.4.3 sex: male and female half and half
14.4.4 number of animals: 50
14.4.5 fasting time: fasting is 12 hours before the experiment
14.5 dosage:
14.5.1 group distance: 0.85
14.5.2 dosage design: according to the trial test result, design 8479.6mg/kg, 7207.5mg/kg, 6126.6mg/kg, 5207.6mg/kg, 4426.4mg/kg tests for 5 dosage groups of test.
14.5.3 approach: per os is irritated stomach
14.6 experimental technique:
Take by weighing arginine and zinc sulfate (ZnSO in (13.47: 1) ratio
2.7H
2O) mix,, be made into desired concn, once irritate stomach by body weight 0.4ml/10g per os and give with 37 ℃ of warm water dissolvings.
14.7 observation index:
14.7.1 the observation period: 14 days
14.7.2 toxic reaction: the NIH its mouse oral gives arginine and zinc sulfate (ZnSO
2.7H
2O) behind the solution, 8479.6mg/kg, 7207.5mg/kg, the animal appearance activity of three dosage groups of 6126.6mg/kg is slowed down, and occurs lying prostrate Bradypneic symptom, most of animal dead after 4～5 hours after 1 hour.5207.6mg/kg two dosage groups of 4426.4mg/kg per os gives arginine and zinc sulfate (ZnSO
4.7H
2O) behind the solution, activity is slowed down a little, and is dead in part animal 48 hours after administration, and all surviving animals major parts were recovered normal diet and activity on the 3rd day after administration, see individual animal death (seeing Table 3) in the observation period, and the no abnormal symptom of animal occurs.
The smart peace of table 3 capsule raw material powder mice LD50 experimental animal death condition
Group number of animals dead animal every day number (only) animal dead sum
(mg/kg) (only) 123 6-14 (my god) (only)
8476.6????10????????9???????????????????????????????9
7207.5????10????????8????1??????????????????????????9
6126.6????10????????4????1??????????????????????????5
5207.6????10????????1???????????????????????????????1
4426.4????10????????0
14.8 result:
Irritate stomach by per os and give smart ammonia capsule raw material powder calculating, LD
50Be 6268.9mg/kg, the 95% credible 5758.2～6825.0mg/kg that is limited to, according to the acute toxicity grading criteria of chemical substance, the nontoxic level in true border.(seeing ∠ D50 report calculated)
LD
50Report calculated 01-3-20 trial drug: smart peace capsule administering mode: irritate stomach dosage group number: 5 algorithms: Bliss method experimental animal: NIH mice dosage unit: mg/kg natural mortality rate: 0.0 precision: 0.001
LD
50Calculations list (the 3rd takes turns)
The moving expection of experiment inflexibly expection work returns and divides the administration logarithm thing dead probability weight of the probability probability probability recurrence group dosage dosage of dying to count unit of digit rate unit and lead the unit coefficient unit error
X??????????????????????????????????????????Y????????????
??????E1???8476.6????3.928??10??9??0.900??6.282??6.844???0.967??5.918??0.168???6.844??0.000362???7207.5????3.858??10??9??0.900??6.282??5.856???0.804??6.203??0.486???5.855??0.000443???6126.5????3.787??10??5??0.500??5.000??4.865???0.446??5.001??0.632???4.865??0.000514???5207.6????3.717??10??1??0.100??3.718??3.874???0.130??3.732??0.396???3.874??0.000595???4426.4????3.646??10??0??0.000???-?????2.883???0.017??2.480??0.107???2.883??0.00066
As scheme shown in the l, with the corresponding curve chart of depicting as of various dose with the laboratory animal mortality rate.The weighted regression equation: the 1st takes turns
(r=0.962 Emax=0.173) the 2nd takes turns=13.933X-47.896
(r=0.945 Emax=0.0201) the 3rd takes turns=14.037X-48.298
=14.038X-48.302 (r=0.943, Emax=0.000663) the heterogeneous check of data: X
2=2.401
P=0.494 (p＞0.05, heterogeneous nonsignificance) regression coefficient standard error: S
b=3.203 return significance coefficient: g=0.200 (the accurate gauged result through Feiller should be adopted in g＞0.10)
LD95％ ( mg/kg ) Feiller Feller％ X Sx LD 95％CL X Sx LD 95％CL1 3.631 0.041 4277.2 3552.7-5149.4 3.590 0.051 3890.8 3097.3-4887.65 3.680 0.031 4783.0 4152.0-5509.9 3.651 0.038 4474.3 3768.5-5312.310 3.706 0.027 5076.7 4503.3-5723.1 3.683 0.032 4820.3 4175.2-5565.150 3.797 0.017 6264.3 5805.7-6759.0 3.797 0.019 6268.9 5758.2-6825.090 3.888 0.027 7729.6 6842.3-8732.0 3.911 0.032 8152.9 7042.7-9438.195 3.914 0.032 8204.2 7105.8-9472.5 3.944 0.039 8783.4 7376.3-10458.899 3.963 0.042 9174.5 7601.8-11072.5 4.004 0.051 10100.5 8015.7-12727.72,FEILLER。 [LD50CALC2.0 version] page 1 (totally 2 pages)
Claims (8)
1, a kind of compound that is used for the treatment of male sterility is characterized in that it mainly is made up of arginine and zinc sulfate, and the ratio of arginine and zinc sulfate is from 4.62: 1 to 19.24: 1.
2, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is from 4.62: 1 to 11: 1.
3, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 13.47: 1 to 19.24: 1.
4, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 11: 1 to 19.24: 1.
5, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 6.6: 1 to 19.24: 1.
6, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 9.44: 1 to 19.24: 1.
7, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 4.62: 1 to 13.47: 1.
8, a kind of compound that is used for the treatment of male sterility according to claim 1, the ratio that it is characterized in that arginine and zinc sulfate is 6.6: 1 to 13.47: 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02115233 CN1197563C (en) | 2002-05-12 | 2002-05-12 | Compound composition for curing male's sterility |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02115233 CN1197563C (en) | 2002-05-12 | 2002-05-12 | Compound composition for curing male's sterility |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1457799A true CN1457799A (en) | 2003-11-26 |
| CN1197563C CN1197563C (en) | 2005-04-20 |
Family
ID=29426478
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02115233 Expired - Lifetime CN1197563C (en) | 2002-05-12 | 2002-05-12 | Compound composition for curing male's sterility |
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| Country | Link |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013163928A1 (en) * | 2012-05-02 | 2013-11-07 | Zhang Fengfan | Complex amino-acid composition and preparation for treating male infertility, impotence and premature ejaculation |
| CN106074598A (en) * | 2016-06-14 | 2016-11-09 | 奥格生物技术（六安）有限公司 | A kind of organic chelated microelement composition of compound recipe treating unusual semen and male infertility disease and preparation method thereof |
| CN106309501A (en) * | 2016-10-18 | 2017-01-11 | 三才石岐制药股份有限公司 | Product for preventing and treating male infertility |
-
2002
- 2002-05-12 CN CN 02115233 patent/CN1197563C/en not_active Expired - Lifetime
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013163928A1 (en) * | 2012-05-02 | 2013-11-07 | Zhang Fengfan | Complex amino-acid composition and preparation for treating male infertility, impotence and premature ejaculation |
| CN106074598A (en) * | 2016-06-14 | 2016-11-09 | 奥格生物技术（六安）有限公司 | A kind of organic chelated microelement composition of compound recipe treating unusual semen and male infertility disease and preparation method thereof |
| CN106309501A (en) * | 2016-10-18 | 2017-01-11 | 三才石岐制药股份有限公司 | Product for preventing and treating male infertility |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1197563C (en) | 2005-04-20 |
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Address after: 528400, Guangdong, Zhongshan Torch Development Zone, Zhongshan port, national health industrial base, 1 Pine Road Patentee after: Guangdong Succhi Pharmaceutical (Group) Ltd. Address before: 528400, No. two, No. 41, Zhongshan Road, Shiqi District, Guangdong, Zhongshan Patentee before: Sancai Medicine Group Co., Ltd., Zhongshan City |
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