CN112972551A - Method for improving mating ability of male Wistar rats - Google Patents
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Abstract
一种提高雄性Wistar大鼠交配能力的方法，属于实验动物生产技术领域，通过采用肉苁蓉、锁阳、淫羊藿和枸杞子四种中药材提取物分别按照0.5mg/mL、1.0 mg/mL、0.6 mg/mL、1.4 mg/mL的浓度配比结合的方式，每日灌胃给雄性Wistar大鼠，使其在短期内交配能力得到明显增强，有效地提高了大鼠捕捉和爬跨次数，显著降低捕捉和爬跨潜伏期，大幅缩短繁殖周期，在实验室鼠类繁育、建立繁殖体系等方面具有成本低、耗时短、操作简单等应用价值，并在濒危哺乳动物繁殖保护、雄性动物勃起功能障碍治疗及开发新药物等方面具有实际的参考依据和广阔的应用前景。
A method for improving the mating ability of male Wistar rats belongs to the technical field of experimental animal production. The combination of 0.6 mg/mL and 1.4 mg/mL concentrations was given to male Wistar rats by gavage every day, so that the mating ability was significantly enhanced in a short period of time, and the number of rats catching and climbing was effectively increased. Significantly reduces the incubation period of capture and climbing, and greatly shortens the breeding cycle. It has low cost, short time-consuming, and simple operation in laboratory mouse breeding and establishment of breeding systems. It has practical reference basis and broad application prospects in the treatment of dysfunction and the development of new drugs.
Description
Technical Field
The invention belongs to the technical field of experimental animal production, relates to a method for improving the mating ability of male rats, and particularly relates to a traditional Chinese medicine extract formula for improving the mating ability of rats.
Background
Wistar rats are white confined group rats, grown in 1907 by the american Wistar (Wistar) institute, and are now distributed throughout the world's laboratories. The main characteristics are as follows: the head is wider, the ears are longer, and the length of the tail is shorter than the body length. Secondly, the sexual cycle is stable, the reproductive capacity is strong, the number of born baby is large, the number of born baby is about 10 per fetus on average, and the growth and development are fast. And thirdly, the resistance to infectious diseases is stronger. Fourthly, the spontaneous tumor incidence is low. The requirement on humidity is very high, and the patient is susceptible to the syndrome of the cricoid when the relative humidity of air is lower than 40 percent.
Wistar rats are the most commonly used species in animal experimental rats and the species used in biomedical research with the longest history. The animals in the population are sensitive to various nutrient substances and medicines, have high absorption rate, sensitive physiological response, obvious behavior change and high stress response speed, and are commonly used in various physiological endocrinology, pharmacology, nutriology and ethology experiments. Although Wistar rats have stable sexual cycle and strong fecundity, the developed method for improving the mating ability of rats and shortening the reproductive cycle has very wide application prospect because the number of rats required by various experiments is large, the price is expensive, the male and female pairing reproductive cycle is long, and the rats for experiments are often in shortage.
According to the existing research, Cistanche deserticola (Cistanches Deserticola) is also called as desert ginseng, is one of traditional Chinese medicinal materials with extremely high medicinal value, has very strong effects of tonifying kidney and strengthening yang, can effectively enhance the activity of sperms and achieves the effects of supplementing marrow and replenishing essence; cynomorium songaricum (Cynomorium songaricum) is commonly called as 'an' medicine, also has high medicinal value, can tonify kidney, replenish vital essence, treat impotence and has certain effect on improving sexual function; epimedium (Epimedium brevicornum) belongs to parasitic herbaceous plants, and leaves of the parasitic herbaceous plants have high medicinal value, and the existing research shows that icariin is a main substance for enhancing the penile erection function, has obvious relaxation effect on the cavernous smooth muscle of the penis of the rabbit, and improves the cGMP level; fructus Lycii is mature fruit of fructus Lycii (Lycium bararum), has obvious effects in resisting fatigue and aging, regulating immunity, resisting tumor, reducing blood lipid and blood sugar, replenishing vital essence, strengthening tendons and bones, improving reproductive capacity, and protecting reproductive system.
According to related reports, the traditional Chinese medicines such as ginseng, pilose antler, desertliving cistanche, cynomorium songaricum, epimedium herb, barbary wolfberry fruit, acanthopanax, common curculigo rhizome, medicinal indianmulberry root and the like can be used as a yang-strengthening medicine for tonifying kidney and replenishing vital essence, so that the sexual desire capability of animals is effectively improved, the conception rate and the litter size of female animals are improved, and the quality of offspring can be improved. However, the traditional Chinese medicinal materials such as ginseng, pilose antler and the like are high in price and many in counterfeit, and are not easy to apply to the production aspect of experimental animals, so that the screening of natural plant traditional Chinese medicinal materials with low price and good drug effect has very important value and significance.
Disclosure of Invention
The invention aims to provide a method for improving mating ability of male Wistar rats according to the physiological characteristics of the Wistar rats and the characteristic that the Wistar rats are sensitive to drugs, and by adopting a mode of combining extracts of four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar according to concentration ratios of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL respectively, stomach irrigation is performed on the male Wistar rats every day, so that the mating ability of the male Wistar rats in a short period is obviously enhanced, the breeding cycle of the rats is shortened, and the method has high laboratory application value.
The technical scheme of the invention is as follows: a method for improving the mating ability of male Wistar rats, which comprises the following steps:
(1) drying four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar, and then crushing the dried Chinese medicinal materials into 150 meshes by using a crusher;
(2) weighing a certain amount of traditional Chinese medicine powder according to the weight ratio of 1: 10, soaking in 70-80% of medical ethanol in a mass ratio, sealing and storing for 24 hours, then performing suction filtration, continuously soaking filter residues in 75% of medical ethanol, repeatedly performing suction filtration for 3 times, and combining filtrates;
(3) concentrating the filtrate under reduced pressure by using a rotary evaporator, setting the temperature at 60-65 ℃, rotating at 60rpm, concentrating to paste, and putting the paste into an oven to evaporate residual moisture;
(4) preparing the extracts of four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar into formula solutions according to the concentration of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL respectively by using distilled water, and feeding the formula solutions to male Wistar rats by intragastric administration every day;
(5) and putting the male rat into a behavior observation box to adapt for 10min, putting the female rat in estrus, observing for 15min, and recording capture times, latency, climbing times and latency.
The concentration of the medical ethanol for soaking in the step (2) is 75%, and the temperature for sealing and storing is 23 +/-2 ℃.
The rotary evaporation temperature in the step (3) is 60-65 ℃; wherein the rotary evaporation temperature of the epimedium and the medlar is 60 ℃; the rotary evaporation temperature of the cynomorium songaricum and the cistanche deserticola is 65 ℃.
The male Wistar rat in the step (4) is 8 weeks old and has the weight of 250-280 g.
The daily gavage dosage in the step (4) is 10 mL/kg.
And (5) observing the behavior in the environment with quiet and dark light.
The invention has the beneficial effects that: the invention provides a method for improving mating ability of male Wistar rats, which is characterized in that extracts of four traditional Chinese medicinal materials including cistanche, cynomorium songaricum, epimedium and wolfberry are respectively combined according to concentration ratios of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL, and the extracts are fed to the male Wistar rats every day, so that the mating ability of the male Wistar rats is obviously enhanced in a short period, the times of catching and climbing of the rats are effectively improved, the catching and climbing latency is obviously reduced, the breeding period is greatly shortened, the method has application values of low cost, short time consumption, simplicity in operation and the like in aspects of laboratory rat breeding, breeding system establishment and the like, and has practical reference basis and wide application prospect in aspects of endangered mammal breeding protection, male erectile dysfunction treatment, new medicine development and the like.
Drawings
FIG. 1 is a graph showing the number of times of capturing male rats in the present invention.
FIG. 2 is a graph showing the capturing latency of male rats in the present invention.
FIG. 3 is a graph showing the number of climbing steps of a male rat in the present invention.
FIG. 4 is a graph showing the climbing latency of male rats in the present invention.
Detailed Description
The invention will be further described with reference to the accompanying drawings in which:
drying four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar, and then crushing the dried Chinese medicinal materials into 150 meshes by using a crusher; weighing a certain amount of traditional Chinese medicine powder according to the weight ratio of 1: 10, soaking in 75% medical ethanol, sealing at 23 +/-2 ℃ for 24 hours, then performing suction filtration, continuously soaking filter residues in 75% medical ethanol, repeatedly performing suction filtration for 3 times, and combining filtrates; concentrating the filtrate under reduced pressure with a rotary evaporator, setting the temperature of Cistanchis herba and herba Cynomorii at 65 deg.C, setting the temperature of herba Epimedii and fructus Lycii at 60 deg.C, rotating at 60rpm, concentrating to extract, and placing in oven to evaporate residual water; preparing the extracts of four Chinese medicinal materials including herba cistanches, herba cynomorii, herba epimedii and fructus lycii into formula solutions according to the concentration of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL respectively by using distilled water; selecting 8 healthy male rats aged 8 weeks and having the weight of 250-280 g, and intragastrically administering the extract formula every day; additionally, a control group (n is 8) is provided, and the normal saline is perfused every day, and the perfused dose is 10 mL/kg. Under a quiet and dark environment, the two groups of male rats are placed into a behavior observation box to adapt for 10min, then placed into a female rat in heat, observed for 15min, and recorded capture times, latency, climbing times and latency.
The cream yield of the four Chinese medicinal material extracts is shown in table 1.
TABLE 1 extraction rate and process of four Chinese medicinal materials
Note: the extract yield is equal to the mass of the extract-like extract/mass of the powdery raw material × 100%.
The number of traps and incubation periods for male rats are shown in figures 1 and 2: compared with a control group, the capturing frequency of male rats in the formula group is obviously increased (p is less than 0.05), and the capturing latency is obviously shortened (p is less than 0.05);
the number of climbs and latency periods for male rats are shown in figures 3 and 4: the climbing times of male rats in the formula group are obviously different from those in the control group (p is less than 0.05), and the climbing latency is greatly shortened (p is less than 0.05);
experiments show that the method for improving the mating ability of male Wistar rats provided by the invention adopts a mode of combining extracts of four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar according to concentration ratios of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL respectively, and the extracts are fed to male rats by intragastric administration every day, so that the mating ability of the male rats in a short period is obviously enhanced; the extract formula adopted is safe, scientific and reasonable, the cost is low, the capture and span times of rats can be effectively improved, the capture and span latency is remarkably reduced, the breeding period is greatly shortened, the application values of low cost, short time consumption, simple operation and the like in the aspects of laboratory rat breeding, breeding system establishment and the like are realized, and the extract has practical reference basis and wide application prospect in the aspects of endangered mammal breeding protection, male animal erectile dysfunction treatment, new medicine development and the like.
Claims (6)
1. A method for improving the mating ability of male Wistar rats, which comprises the following steps:
(1) drying four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar, and then crushing the dried Chinese medicinal materials into 150 meshes by using a crusher;
(2) weighing a certain amount of traditional Chinese medicine powder according to the weight ratio of 1: 10, soaking in 70-80% of medical ethanol in a mass ratio, sealing and storing for 24 hours, then performing suction filtration, continuously soaking filter residues in 75% of medical ethanol, repeatedly performing suction filtration for 3 times, and combining filtrates;
(3) concentrating the filtrate under reduced pressure by using a rotary evaporator, setting the temperature at 60-65 ℃, rotating at 60rpm, concentrating to paste, and putting the paste into an oven to evaporate residual moisture;
(4) preparing the extracts of four Chinese medicinal materials of cistanche, cynomorium songaricum, epimedium and medlar into formula solutions according to the concentration of 0.5mg/mL, 1.0mg/mL, 0.6mg/mL and 1.4mg/mL respectively by using distilled water, and feeding the formula solutions to male Wistar rats by intragastric administration every day;
(5) and putting the male rat into a behavior observation box to adapt for 10min, putting the female rat in estrus, observing for 15min, and recording capture times, latency, climbing times and latency.
2. The method of claim 1, wherein the method comprises the steps of: the concentration of the medical ethanol for soaking in the step (2) is 75%, and the temperature for sealing and storing is 23 +/-2 ℃.
3. The method of claim 1, wherein the method comprises the steps of: the rotary evaporation temperature in the step (3) is 60-65 ℃; wherein the rotary evaporation temperature of the epimedium and the medlar is 60 ℃; the rotary evaporation temperature of the cynomorium songaricum and the cistanche deserticola is 65 ℃.
4. The method of claim 1, wherein the method comprises the steps of: the male Wistar rat in the step (4) is 8 weeks old and has the weight of 250-280 g.
5. The method of claim 1, wherein the method comprises the steps of: the daily gavage dosage in the step (4) is 10 mL/kg.
6. The method of claim 1, wherein the method comprises the steps of: and (5) observing the behavior in the environment with quiet and dark light.
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| CN114191569A (en) * | 2021-12-19 | 2022-03-18 | 扬州大学 | A kind of preparation method of nanometer medicine based on hydroxypropyl-β-cyclodextrin |
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| CN1113445A (en) * | 1995-02-06 | 1995-12-20 | 李忠 | Health-care medicine composition and its production |
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| CN114191569A (en) * | 2021-12-19 | 2022-03-18 | 扬州大学 | A kind of preparation method of nanometer medicine based on hydroxypropyl-β-cyclodextrin |
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