CN102845365A - Modeling method of uyghur medicine animal model, model, and disproof and comprehensive evaluation method - Google Patents
Modeling method of uyghur medicine animal model, model, and disproof and comprehensive evaluation method Download PDFInfo
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Abstract
The invention discloses a modeling method of an uyghur medicine abnormal balgam syndrome and disease animal model, the model, and a disproof and comprehensive evaluation method, and belongs to the field of uyghur medicine andrology. The modeling method of the abnormal balgam syndrome and animal model is characterized in that after the model is formed, regrouping, disproof and comprehensive evaluation are performed. The method comprises the following steps: dividing experimental animals into a control group and a modeling group; breeding the control group with conventional diet in a conventional environment; placing the modeling group into an artificial climate box (temperature is set to 6 DEG C and humidity is set to 85%-95%), and breeding the modeling group with wet and cold feed, while both the control group and the modeling groups drink water randomly; and dynamically observing biological representation, penis erection function and sexology change of each group in the whole course. The modeling method of the uyghur medicine animal model, the model, and the disproof and comprehensive evaluation method provided by the invention has reliability, feasibility and favorable repetitiveness and scientificity, and can be used for biological fundamental research and pharmaceutical intervention research of uyghur medicine abnormal balgam syndrome and abnormal balgam impotence disease.
Description
Technical field
The invention belongs to Uygur's medical science andrological diseases research field, be specifically related to a kind of modeling method of the unusual mucus cross-examination marquis animal model of doctor, a kind of unusual mucus cross-examination marquis model of doctor, a kind of counterevidence and integrated evaluating method of tieing up the unusual lymphatic temperament impotence disease animal model preparation of doctor tieed up tieed up.
Background technology
Impotence refers to man's penis erectile problem or act and not hardly is the disease of principal character, is called Erectile Dysfunction (erectile dysfunction, ED) in modern medicine, and visible sexual desire lowers and the card shapes such as premature ejaculation.The society male sex accelerates because of rhythm of life, and the average length of one's sleep, few, dietary structure and environmental evolution more caused the incidence of disease of impotence to rise year by year because of mental health and other kinds paathogenic factor.The modern medicine study data shows and worldwide to reach 1.52 hundred million people in nineteen ninety-five ED patient, in the male sex in 40-70 year, approximately has 52% the male sex that in various degree ED is arranged, and expects 2025 and will rise to 3.22 hundred million people; China's city male sicken rate reaches that total prevalence rate is 40.2% more than 26.1%, 40 years old.At present for ED medicine and method, although there are the several different methods such as V-type Pimobendane, hormone, AGEs inhibitor available, especially the generation of phosphodiester bond inhibitor has brought revolutionary variation to the treatment of ED, yet still have larger gap with people's expected value, the wholistic therapy effect still has some deficits.
Uygur's medical science plays an important role in the health care system of Xinjiang as the important component part of motherland's traditional medicine.It has distinctive feature and significant curative effect with theoretical system independently and unique methods for the treatment of in the treatment of andrological diseases dimension medical science, is dimension doctor's a large advantage characteristic to the treatment of impotence, premature ejaculation class disease especially.But start late because of dimension doctor experimental zoology, the disease animal model of different syndrome type impotence is not yet set up, and causes the pathogenetic research of its Basic of Biology and modern medicine still to belong to blank, thereby is restricting the standardization and modernization of the diagnosis and treatment of dimension doctor andrology.In this simultaneously, the work such as the experimental study on disease animal model basis and new drug development, evaluating drug effect can't be carried out, and therefore set up the unusual lymphatic temperament impotence disease animal model of dimension doctor.
Summary of the invention
For above-mentioned the deficiencies in the prior art, one of purpose of the present invention provides a kind of modeling method of tieing up the unusual mucus cross-examination marquis animal model of doctor, and the method can be used and set up the unusual mucus cross-examination marquis model of dimension doctor.
Technical scheme is: a kind of modeling method of tieing up the unusual mucus cross-examination marquis animal model of doctor, and the method is as follows: laboratory animal is divided into control group and modeling group; Control group is raised in conventional environment, every group every day 20:00-22:00 to the 300g normal diet, 500 ml water are changed bedding and padding, the bedding and padding amount is 80 g; The modeling group places climatic cabinate to raise, and every group of every day, 20:00-22:00 gave raw feed 300g, and 500 ml water are changed bedding and padding, and the bedding and padding amount is 80 g; Control group and modeling group be random diet water all; Dynamically observation.
As preferably, described laboratory animal is rat.
As preferably, described rat is male SD rat.
Two of purpose of the present invention provides a kind of dimension and cures unusual mucus cross-examination marquis model, and this model can be applicable to tie up counterevidence and the integrated evaluating method of the unusual lymphatic temperament impotence disease animal model preparation of doctor.
Technical scheme is: a kind of dimension is cured unusual mucus cross-examination marquis model, and the method adopts the described dimension of the arbitrary claim of the claims 1-3 to cure the modeling method of unusual mucus cross-examination marquis animal model.
Three of purpose of the present invention provides a kind of counterevidence and integrated evaluating method of tieing up the unusual lymphatic temperament impotence disease animal model preparation of doctor, and the method can be applicable in the clinical testing of impotence disease.
Technical scheme is: a kind of counterevidence and integrated evaluating method of tieing up the unusual lymphatic temperament impotence disease animal model preparation of doctor, and the method is as follows: set up dimension as claimed in claim 4 and cure unusual mucus cross-examination marquis model; Select the impotence model of a syndrome by APO telotism experiment and associativity behaviouristics testing sieve; The impotence model of a syndrome is divided into Syndrome model group A1, natural counterevidence group A2 and medicine counterevidence group A3 at random; Do not become the card marquis model of impotence to be divided at random card marquis model group B1, natural counterevidence group B2 and medicine counterevidence group B3, the counterevidence time was 2 weeks; When counterevidence finishes, carry out again APO telotism experiment and sexology and detect; Naturally counterevidence contrast: nature counterevidence group A2 and B2 were raised for 2 weeks under conventional environment, regular diet, by Biological Characterization observation and with Normal group relatively after, determine the natural recovery situation of model; Medicine counterevidence contrast: medicine counterevidence group A3 and B3 were raised for 2 weeks under conventional environment, and regular diet intervened for 2 weeks with verification medicine Yimusake table simultaneously, and by Biological Characterization observation and with Normal group relatively after, verify the result for the treatment of of medicine; Sexology detects: indoor keeping quite, light only is dimmed to and enough observes, male rat is placed on conforms 10 minutes in the glass box, drop into female mouse by 2:1, observe and record riding latent period, ride number of times, insert latent period, inserting number of times, ejaculation latency, ejaculation frequency of male rat in 30 minutes; Described regular diet be every group every day 20:00-22:00 to the 300g normal diet, 500 ml water.
As preferably, described medicine counterevidence group A3 and B3 every day are with 250mg Yimusake table/1kg distilled water gavage, and natural counterevidence group A2, natural counterevidence group B2, Syndrome model group A1 and card marquis model group B1 every day are with 1kg distilled water gavage.
As preferably, described APO telotism is placed on male rat first and conforms 10 minutes in the inspection box, light dims, only enough the observation gets final product, indoor keeping quite, then every rat is at neck cutis laxa place injection apomorphine (APO) 85 μ g/kg (APO is dissolved in the vitamin C and physiological saline of 0.5mg/kg, and the adjustment volume is 5ml/kg), every rat was observed 30 minutes after injection at once, and observation index comprises erect latent period and erection number of times.Glans penis hyperemia and terminal phallosome telotism occurs being designated as once.
As preferably, described Biological Characterization is observed qualitative index observation and quantitative target is observed.
As preferably, described qualitative index is observed skin and hair and observes under daylight, emotional reactions each rat in the cage reacts to each other as subjectivity and examines 30 minutes, the activity of behavior state in during this observed 30 minutes under main, quiet environment, reaction when excitement degree is tested take the folder tail is as main, the tongue picture tongue fur is observed, records and is taken pictures under daylight, the diet water state is organized the same time and is fed to raise and observe the diet water state with each; when urine was just urinated take each group rat the same day color as main, the stool state during take the same day positive bowel movement the stool form as main;
Described quantitative target is for avoiding circadian impact, be fixed in 20:00-22:00 every day and collect every data, body weight wherein: weigh in required time and record every day, with every cage TBW divided by this cage rat number of elements, draw average weight, and draw model group each cage body weight ratio and record, the initial body weight of the body weight ratio=model group average weight-model group/initial body weight of normal group average weight-normal group by following formula; Dietary amount: every day, 20:00 added feed 300g, and next day, 20:00 claimed the food surplus, drew the corresponding dietary amount of 100g body weight and record according to following formula; Amount of drinking water: 20:00 feedwater every day 500ml, next day, 20:00 amount surplus water calculated the corresponding amount of drinking water of 100g body weight and record; The urine amount: the modeling first day record to dried dunnage weight, second day claims to contain weight and the record of the wet bedding and padding of urine and stool in required time, then again weighs after 100 ℃ of oven dry in lower 2 hours of constant temperature blast drying oven, calculates the urine amount of respectively organizing; Stool amount: calculate corresponding stool amount and the record of 100g body weight.
Find by experiment of the present invention, can recover to give counterevidence and verify the understanding characteristics that the unusual mucus cross-examination marquis that sets up medicine counterevidence aspect and impotence disease rat model meet the unusual mucus cross-examination of Uygur's medical science marquis from the simulation cause of disease, card shape performance, chronic process and nature, the general state change that simulates meets the unusual mucilagenous card Hou Tezheng of Uygur's medical science, the ED model of screening, also more stable.We think, unusual mucus cross-examination marquis and the impotence disease animal model set up in this research are reliable, feasible, and have good repeatability and science.
The animal model of the unusual lymphatic temperament impotence of the present invention disease can be used for unusual mucus cross-examination and unusually Basic of Biology research and the pharmaceutical intervention research of lymphatic temperament impotence disease.
Beneficial effect of the present invention is: the present invention is by taking raw environment and raw feed to raise the unusual lymphatic temperament impotence disease animal model that obtains.Data shows, the modeling group has occurred curling up sleeping few moving, and happiness flocks together, and is slow in action, and hair is disorderly matt, and dark tongue quality, white and greasy fur, oral cavity, nasal secretion such as increase at the phenomenon.Simultaneously visible modeling group body weight gain is slow, dietary amount increases, amount of drinking water reduces, the stool and urine amount significantly increases, and urine is clear, and look light, the situation such as half congealed when soft during stool, above-mentioned Biological Characterization meets the clinical card Hou Tedian of dimension doctor, thus from unusual mucus cross-examination marquis rat model successfully screening impotence disease rat model, and this model has good science, reliability and stability.
Embodiment
Be that the present invention is described further below in conjunction with subordinate list.
The preparation of the unusual lymphatic temperament impotence disease animal model of dimension doctor:
(1) laboratory animal SD rat, average weight 203-237g, cleaning level SPF level, Xinjiang Medicine University's Experimental Animal Center provides.
(2) main agents, medicine and experimental instrument: apomorphine, U.S. Sigma company; Progesterone, Zhejiang Province XianJu Pharmacy stock Co., Ltd; Estradiol, Shanghai General Pharmaceutical Co., ltd.; RQH-350 type climatic cabinate, the grand experimental facilities of upper Nereid Co., Ltd product; Yimusake table, medicine company Co., Ltd of hotan Uygur.
(3) experimental technique: 100 male SD rats with normal sexual function, all in conventional environment, raised in 1-3 days before the experiment.Therefrom get at random 10 for Normal group, in conventional environment, raise by (20 ℃ ± 2 ℃ of temperature, humidity 55%-65%, 12h/12h artificial illumination), every day the 300g normal diet, 500ml water, 80 bedding and padding; All the other 90 are the modeling group, place climatic cabinate to raise (6 ℃ of design temperatures, humidity 85%-95%), 9:00-19:00 puts into, the raw feed of 300g every day, this raw feed is made by adding real each 150g of coriandri,fructus and spinach in every 1kg conventional feed, and water is identical with normal group with bedding and padding.Fixedly every day, 20:00-22:00 gave feed, drinking-water, changed bedding and padding, the variation of observation Biological Characterization.
The directed index observing of table one, the unusual mucilaginous substance impotence disease animal model of dimension doctor.
The body weight of unusual lymphatic temperament impotence disease animal model and body weight ratio change.
The variation of table two, body weight average (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 222.97±8.42 | 377.97±8.66 | 479.24±6.11 | 553.79±7.00 | 631.66±2.89 | 662.57±11.51 |
| The modeling group | 229.20±7.81 | 333.48±14.83 ﹡ | 403.13±16.77 ﹡ | 465.59±18.39 ﹡ | 509.49±18.77 ﹡ | 528.70±14.78 ﹡ |
Normal group and modeling group body weight average comparison no difference of science of statistics before the modeling (
P0.05); The 4th thoughtful the 20th week of modeling, modeling group body weight average be lower than normal group (
P＜0.05).
The variation of table three, body weight ratio (
)
| Grouping | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 0.98±0.02 | 1.00±0.01 | 1.01±0.01 | 1.00±0.00 | 1.02±0.02 |
| The modeling group | 0.68±0.10 ﹡ | 0.68±0.01 ﹡ | 0.72±0.05 ﹡ | 0.69±0.04 ﹡ | 0.72±0.04 ﹡ |
Modeling group body weight ratio also significantly less than normal group (
P＜0.05).
The urine amount of unusual lymphatic temperament impotence disease animal model and stool quantitative change:
The dietary amount and the amount of drinking water that are converted into 100g body weight rat through formula compare.
The variation of table four, modeling stage 100g body weight dietary amount (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 17.81±2.69 | 8.69±0.51 | 7.39±0.56 | 7.80±0.58 | 7.61±0.41 | 7.71±0.42 |
| The modeling group | 16.65±1.75 | 11.85±0.97 | 10.41±0.97 | 10.63±1.12 | 10.68±0.15 | 10.25±0.57 |
The dietary amount difference not statistically significant of normal group and modeling group before the modeling (
P0.05); Modeling stage modeling group dietary amount all greater than normal group (
P＜0.05).
The variation of table five, modeling stage 100g body weight amount of drinking water (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 16.19±2.11 | 11.94±0.58 | 11.12±1.11 | 11.45±1.33 | 10.59±0.63 | 10.40±0.29 |
| The modeling group | 16.04±2.71 | 11.31±1.98 | 10.22±1.55 ﹡ | 9.89±1.26 ﹡ | 8.83±0.76 ﹡ | 8.91±0.70 ﹡ |
Amount of drinking water difference not statistically significant between modeling front 8 all modeling groups and normal group (
P0.05); The the 8th the thoughtful the 20th all modeling group amounts of drinking water all significantly be less than normal group (
P＜0.05).
The urine amount of unusual lymphatic temperament impotence disease animal model and stool quantitative change:
The urine amount and the stool amount that are converted into 100g body weight rat through formula compare.
The variation of table six, modeling stage 100g body weight urine amount (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 3.18±0.44 | 2.81±0.43 | 3.68±0.57 | 3.75±0.47 | 4.18±0.41 | 4.37±0.30 |
| The modeling group | 3.66±0.63 | 6.94±1.51 ﹡ | 7.57±1.19 ﹡ | 8.01±1.46 ﹡ | 10.95±0.58 ﹡ | 10.12±0.54 ﹡ |
Modeling group and normal group urine amount no difference of science of statistics before the modeling (
P0.05); Modeling stage modeling group urine amount more than normal group (
P＜0.05).
The variation of table seven, modeling stage 100g body weight stool amount (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 3.24±0.62 | 2.82±0.36 | 2.13±0.17 | 2.49±0.22 | 2.58±0.40 | 2.43±0.20 |
| The modeling group | 2.75±0.52 | 4.77±0.90 ﹡ | 4.18±0.47 ﹡ | 4.14±0.62 ﹡ | 3.93±0.84 ﹡ | 3.33±0.64 ﹡ |
Modeling group and normal group stool amount no difference of science of statistics before the modeling (
P0.05); Modeling stage modeling group stool amount more than normal group (
P＜0.05).
The APO of unusual lymphatic temperament impotence disease animal model erects and tests:
After raising for 20 weeks, first male rat is placed on and conforms 10 minutes in the inspection box, light dims, only enough the observation gets final product, indoor keeping quite, then every rat (APO is dissolved in the vitamin C and physiological saline of 0.5mg/kg at neck cutis laxa place injection apomorphine 85 μ g/kg, the adjustment volume is 5ml/kg), every rat was observed 30 minutes after injection at once, observed erect latent period and erection number of times (glans penis hyperemia and terminal phallosome telotism occurs being designated as once).
Table eight, modeling stage erect latent period relatively (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 465.40±264.57 | 432.40±204.03 | 549.00±201.56 | 501.30±125.78 | 507.00±153.18 | 479.30±143.41 |
| Model group | 440.43±240.69 | 604.43±365.43 | 1107.50±502.03 * | 1306.85±553.37 * | 1361.38±562.31 * | 1381.43±536.81 * |
Table nine, modeling stage erection number of times relatively (
)
| Grouping | Before the modeling | The 4th week | The 8th week | The 12nd week | The 16th week | The 20th week |
| Normal group | 1.60±0.69 | 1.80±0.91 | 2.10±1.19 | 2.10±0.87 | 1.80±0.92 | 1.90±0.74 |
| Model group | 1.45±0.56 | 1.51±0.62 | 0.86±0.65 * | 0.51±0.59 * | 0.46±0.63 * | 0.49±0.67 * |
Difference that there are no significant between modeling group rat erection latent period and erection number of times and normal group before the modeling (
P0.05); Front 8 all modeling group rat erection latent period of modeling stages and erection number of times and normal group difference no difference of science of statistics (
P0.05), the 8th thoughtful the 20th week is with the prolongation of modeling time, modeling group erection number of times compared with normal group reduces, erect gradually to prolong latent period, and group difference all have statistical significance (
P＜0.05), reaches 56.7% to the complete impotence rate of the 20th all modeling groups.
The counterevidence of unusual lymphatic temperament impotence disease animal model:
After the experiment of erecing through APO, filter out the impotence model of a syndrome, again it is divided at random Syndrome model group A1, natural counterevidence group A2 and medicine counterevidence group A3; Do not become the card marquis model of impotence to be divided at random again card marquis model group B1, natural counterevidence group B2, medicine counterevidence group B3; The counterevidence time was 2 weeks.
Naturally counterevidence: divide into groups complete after, the nature counterevidence organized under normal condition raised for 2 weeks, regular diet, and by Biological Characterization observation and with Normal group relatively after, determine the natural recovery situation of model.
Medicine counterevidence: divide into groups complete after, the medicine counterevidence organized under normal condition raise, regular diet, and intervened for 2 weeks with verification medicine Yimusake table, namely give Yimusake table 250mg/kg gavage, and by Biological Characterization observation and with Normal group relatively after, the result for the treatment of of verification medicine.
(1) observation of directed index of counterevidence stage.
(2) the biological variation that characterizes of counterevidence stage.
Give counterevidence the 1st all comparative results (
)
| Grouping | The body weight average | Body weight ratio | 100g body weight dietary amount | 100g body weight amount of drinking water | 100g body weight urine amount | 100g body weight stool amount |
| N | 671.10±12.25 | 1.07±0.13 | 8.03±0.19 | 10.47±0.27 | 4.79±0.41 | 2.47±0.26 |
| A 1 | 546.40±15.95 | 0.18±0.20 | 9.68±0.51 | 8.75±0.52 | 10.18±0.31 | 3.00±0.29 |
| A 2 | 541.29±1.83 | 0.32±0.15 | 9.23±0.53 | 8.46±0.46 | 9.31±0.51 | 3.19±0.45 |
| A 3 | 528.81±4.70 | 0.42±0.24 | 9.76±0.47 | 9.02±0.37 | 9.73±0.62 | 2.77±0.32 |
| B 1 | 533.81±2.79 | 0.20±0.12 | 10.13±0.44 | 9.32±0.45 | 9.84±0.69 | 3.85±0.60 |
| B 2 | 528.43±7.08 | 0.28±0.23 | 10.24±0.59 | 8.72±0.57 | 9.51±0.48 | 3.80±0.62 |
| B 3 | 525.81±7.77 | 0.89±0.21 | 10.06±0.55 | 9.32±0.67 | 10.12±0.61 | 3.80±0.33 |
Body weight and body weight ratio, each model group body weight average of counterevidence stage all significantly be lower than normal group (
P＜0.05), B
1, B
2, B
3The body weight average all is lower than A
1And A
2, group difference all have significant difference (
P＜0.05); Body weight ratio result shows, give counterevidence the 1st week each model group body weight gain all be less than normal group (
P＜0.05);
Dietary amount and amount of drinking water, give counterevidence the 1st the week each model group 100g body weight dietary amount all significantly more than normal group (
P＜0.05), A wherein
2Group is greater than A
1Group (
P＜0.05), B
1, B
2, B
3The difference not statistically significant (
P0.05);
Urine amount and stool amount, each model group urine amount of counterevidence stage all significantly more than normal group (
P＜0.05), the 1st all A
2, A
3The urine amount significantly is less than A
1Group (
P＜0.05), B
1, B
2, B
3The difference not statistically significant (
P0.05).
Gave counterevidence for the 2nd weekComparative result (
)
| Grouping | The body weight average | Body weight ratio | 100g body weight dietary amount | 100g body weight amount of drinking water | 100g body weight urine amount | 100g body weight stool amount |
| N | 678.59±12.92 | 1.08±0.11 | 7.66±0.30 | 10.45±0.25 | 5.09±0.19 | 2.85±0.23 |
| A 1 | 549.11±15.04 | 0.25±0.10 | 9.99±0.28 | 8.44±0.40 | 10.02±0.32 | 3.19±0.19 |
| A 2 | 550.82±5.06 | 0.68±0.25 | 9.37±0.38 | 8.74±0.38 | 9.05±0.46 | 3.01±0.36 |
| A 3 | 540.45±6.58 | 0.86±0.31 | 9.13±0.60 | 9.25±0.40 | 8.40±0.37 | 2.56±0.30 |
| B 1 | 539.20±3.21 | 0.40±0.12 | 10.30±0.26 | 8.78±0.56 | 9.74±0.37 | 3.40±0.45 |
| B 2 | 539.53±9.34 | 0.74±0.27 | 9.19±0.30 | 9.89±0.65 | 8.77±0.62 | 2.65±0.30 |
| B 3 | 537.73±11.00 | 1.17±0.28 | 8.91±0.38 | 10.00±0.32 | 8.60±0.52 | 2.66±0.32 |
Body weight and body weight ratio, the 2nd all B give counterevidence
3Weight gain and normal group difference not statistically significant (
P0.05), remaining each the group still be less than normal group (
P＜0.05);
Dietary amount and amount of drinking water, give counterevidence the 2nd the week each model group amount of drinking water still significantly be less than normal group (
P＜0.05), A
3Than A
1, A
2The group significantly increase (
P＜0.05), B
2, B
3Group is also significantly more than B
1Group (
P＜0.05);
Urine amount and stool amount the 2nd all A that give counterevidence
1, B
1Organize the stool amount all more than normal group, A
3Group less and normal group, difference all have statistical significance (
P＜0.05), B
2, B
3Group stool amount all is less than B
1Group (
P＜0.05).
(3) counterevidence stage A PO experiment.
The counterevidence before and after respectively organize the APO Comparison of experiment results (
)
After counterevidence finishes, A
1, A
2Exist between latent period, erection number of times and the normal group of erecing significant difference (
P＜0.05), remaining each model group aspect erect latent period and erection number of times with the normal equal not statistically significant of group difference (
P0.05), A
1And A
2Between also there was no significant difference (
P0.05), A
3With A
1, A
2Group is more then erected and is significantly shortened latent period, the erection number of times significantly increase (
P＜0.05), B
1, B
2, B
3Between erect latent period and erection number of times the equal not statistically significant of difference (
P0.05).
When counterevidence finished, the sexology of going again detected.
The preparation of heat mouse: female rats is anaesthetized with 1% yellow Jackets (50 mg/ kg), and the abdomen position is fixed.Sterilization skin carries out Bilateral oophorectomy.Postoperative intramuscular injection penicillin 20,000 U/kg-1.d-1 inject 7d altogether.Rested for 2 weeks, test the female alcohol of front 48 h hypodermic injection benzoic acid (20ug/ is only), test front 24 h and again inject the female alcohol of same dose benzoic acid, 4h hypodermic injection progesterone before the experiment, 500ug/, the female alcohol of benzoic acid, progesterone are dissolved in the sesame oil.
Sexology detects: indoor keeping quite, light dims, only enough the observation gets final product, male rat is placed on 10 min that conform in the glass box, drop into female mouse by 2:1, observation is also recorded riding latent period, riding number of times of male rat in 30 minutes, inserts latent period, inserts number of times, ejaculation latency, ejaculation frequency etc.
Table ten, modeling stage mating test result relatively (
)
With normal group than * P＜0.05
Give counterevidence after 2 weeks, ride latent period, insert latent period and ejaculation latency A1, A2, B1 group all significantly are longer than normal group (P＜0.05), ride, insertion, ejaculation frequency also significantly be less than normal group (P＜0.05).A3 group than A1, A2 group insert latent period, ejaculation latency significantly shortens (P＜0.05), inserts number of times, ejaculation frequency significantly increases (P＜0.05); Relatively, insert and significantly shorten with ejaculation latency latent period between B3 and B1 group, ride number of times, insertion number of times, ejaculation frequency and significantly increase (P＜0.05).Simultaneously as seen, ride on number of times and the ejaculation frequency that B2 organizes and there is significant difference (P＜0.05) in B1 between organizing.The equal not statistically significant of difference on indices of 2 groups of A1 and A, B2 and B3 group (P〉0.05).
Table ten one, counterevidence respectively organize after 2 weeks the mating test result relatively (
)
| Grouping | Ride latent period | Ride number of times | Insert latent period | Insert number of times | Ejaculation latency | Ejaculation frequency |
| N | 426.60±143.59 | 29.20±9.55 | 512.60±181.28 | 21.90±7.49 | 559.20±164.11 | 11.70±4.42 |
| A 1 | 1101.08±770.89 * | 6.50±7.40 * | 1800.00±0.00 * | 0.00±0.00 * | 1800.00±0.00 * | 0.00±0.00 * |
| A 2 | 955.25±555.17 * | 10.83±9.31 * | 1491.75±489.38 * | 4.08±7.17 * | 1511.83±468.88 * | 2.42±4.60 * |
| A 3 | 890.58±737.89 | 13.75±16.00 * | 1087.50±715.86 *# | 9.75±12.65 *# | 1098.17±703.53 *#◆ | 5.92±7.90 *# |
| B 1 | 910.55±608.40 * | 9.73±11.55 * | 1116.27±670.31 *# | 6.55±9.07 * | 1149.64±645.62 *# | 3.73±5.64 * |
| B 2 | 465.27±222.49 #◆ | 20.82±10.99 #▽ | 732.27±579.27 #◆ | 14.18±10.53 #◆ | 787.91±554.41 #◆▲ | 10.45±8.51 #▽ |
| B 3 | 480.00±231.79 #◆ | 24.80±10.57 #◆▲▽ | 589.60±234.21 #◆▲▽ | 18.50±9.76 #◆▲▽ | 600.70±236.32 #◆▲▽ | 12.60±6.60 #◆▽ |
With N than * P＜0.05; With A1 than #P＜0.05; With the A2 ratio ◆ P＜0.05; With A3 ratio ▲ P＜0.05; With B1 than ▽ P＜0.05
Result of study shows, can recover to give counterevidence and verify the understanding characteristics that the unusual mucus cross-examination marquis that sets up medicine counterevidence aspect and impotence disease rat model meet the unusual mucus cross-examination of Uygur's medical science marquis from the simulation cause of disease, card shape performance, chronic process and nature, the general state change that simulates meets the unusual mucilagenous card Hou Tezheng of Uygur's medical science, the ED model of screening, also more stable.Unusual mucus cross-examination marquis and the impotence disease animal model set up in this research are reliable, feasible, and have good repeatability with scientific; This can be the Basic of Biology research of unusual mucus cross-examination and unusual mucilaginous substance impotence disease and pharmaceutical intervention research good animal model is provided.
Claims (9)
1. tie up the modeling method of curing unusual mucus cross-examination marquis animal model for one kind, the method is as follows:
Laboratory animal is divided into control group and modeling group;
Control group is raised in conventional environment, every group every day 20:00-22:00 to the 300g normal diet, 500 ml water are changed bedding and padding, the bedding and padding amount is 80 g; The modeling group places climatic cabinate to raise, 6 ℃ of design temperatures, and humidity 85%～95%, every group of every day, 20:00～22:00 gave raw feed 300g, and 500 ml water are changed bedding and padding, and the bedding and padding amount is 80 g;
Control group and modeling group be random diet water all;
Dynamically observation.
2. dimension according to claim 1 is cured the modeling method of unusual mucus cross-examination marquis animal model, it is characterized in that: described laboratory animal is rat.
3. dimension according to claim 2 is cured the preparation method of unusual lymphatic temperament impotence disease animal model, it is characterized in that: described rat is male SD rat.
4. a dimension is cured unusual mucus cross-examination marquis model, it is characterized in that: the method adopts the described dimension of the arbitrary claim of the claims 1-3 to cure the modeling method of unusual mucus cross-examination marquis animal model.
5. tie up counterevidence and the integrated evaluating method that the unusual lymphatic temperament impotence disease animal model of doctor prepares for one kind, the method is as follows:
Set up dimension as claimed in claim 4 and cure unusual mucus cross-examination marquis model;
Filter out the impotence model of a syndrome by APO telotism experiment and the detection of associativity behaviouristics;
The impotence model of a syndrome is divided into Syndrome model group A1, disease nature counterevidence group A2 and disease medicine counterevidence group A3 at random; Do not become the card marquis model of impotence to be divided at random card marquis model group B1, card Hou Ziran counterevidence group B2 and card marquis medicine counterevidence group B3, the counterevidence time was 2 weeks;
When counterevidence finishes, carry out again APO telotism experiment and sexology and detect;
Naturally counterevidence contrast: nature counterevidence group A2 and B2 were raised for 2 weeks under conventional environment, regular diet, by Biological Characterization observation and with Normal group relatively after, determine the natural recovery situation of model;
Medicine counterevidence contrast: medicine counterevidence group A3 and B3 were raised for 2 weeks under conventional environment, regular diet, simultaneously intervened for 2 weeks with verification medicine Yimusake table, and by Biological Characterization observation and with Normal group relatively after, determine the result for the treatment of of verification medicine;
Sexology detects: indoor keeping quite, light only is dimmed to and enough observes, male rat is placed on conforms 10 minutes in the glass box, drop into female mouse by 2:1, observe and record riding latent period, ride number of times, insert latent period, inserting number of times, ejaculation latency, ejaculation frequency of male rat in 30 minutes;
Described regular diet be every group every day 20:00-22:00 to the 300g normal diet, 500 ml water.
6. a dimension as claimed in claim 5 is cured counterevidence and the integrated evaluating method that unusual lymphatic temperament impotence disease animal model prepares, it is characterized in that: described medicine counterevidence group A3 and B3 are with 250mg Yimusake table/1kg distilled water gavage every days, and natural counterevidence group A2, natural counterevidence group B2, Syndrome model group A1 and card marquis model group B1 every day are with 1kg distilled water gavage.
7. a dimension as claimed in claim 5 is cured counterevidence and the integrated evaluating method that unusual lymphatic temperament impotence disease animal model prepares, it is characterized in that: described APO telotism is placed on male rat first and conforms 10 minutes in the inspection box, light dims, only enough the observation gets final product, indoor keeping quite, then every rat (APO is dissolved in the vitamin C and physiological saline of 0.5mg/kg at neck cutis laxa place injection apomorphine (APO) 85 μ g/kg, the adjustment volume is 5ml/kg), every rat was observed 30 minutes after injection at once, and observation index comprises erect latent period and erection number of times; Glans penis hyperemia and terminal phallosome telotism occurs being designated as once.
8. a dimension as claimed in claim 5 is cured counterevidence and the integrated evaluating method that unusual lymphatic temperament impotence disease animal model prepares, and it is characterized in that: described Biological Characterization is observed qualitative index observation and quantitative target observation.
9. a dimension as claimed in claim 7 is cured counterevidence and the integrated evaluating method that unusual lymphatic temperament impotence disease animal model prepares, and it is characterized in that:
Described qualitative index is observed skin and hair and observes under daylight, emotional reactions each rat in the cage reacts to each other as subjectivity and examines 30 minutes, the activity of behavior state in during this observed 30 minutes under main, quiet environment, reaction when excitement degree is tested take the folder tail is as main, the tongue picture tongue fur is observed, records and is taken pictures under daylight, the diet water state is organized the same time and is fed to raise and observe the diet water state with each; when urine was just urinated take each group rat the same day color as main, the stool state during take the same day positive bowel movement the stool form as main;
Described quantitative target is for avoiding circadian impact, be fixed in 20:00-22:00 every day and collect every data, body weight wherein: weigh in required time and record every day, with every cage TBW divided by this cage rat number of elements, draw average weight, and draw model group each cage body weight ratio and record, the initial body weight of the body weight ratio=model group average weight-model group/initial body weight of normal group average weight-normal group by following formula; Dietary amount: every day, 20:00 added feed 300g, and next day, 20:00 claimed the food surplus, drew the corresponding dietary amount of 100g body weight and record according to following formula; Amount of drinking water: 20:00 feedwater every day 500ml, next day, 20:00 amount surplus water calculated the corresponding amount of drinking water of 100g body weight and record; The urine amount: the modeling first day record to dried dunnage weight, second day claims to contain weight and the record of the wet bedding and padding of urine and stool in required time, then again weighs after 100 ℃ of oven dry in lower 2 hours of constant temperature blast drying oven, calculates the urine amount of respectively organizing; Stool amount: calculate corresponding stool amount and the record of 100g body weight.
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| CN106645748A (en) * | 2016-12-16 | 2017-05-10 | 新疆医科大学 | Mammalian differential serum protein and differentially expressed protein system of penile smooth muscle tissue of abnormal and phlegmatic type of impotence syndrome and screening method and application |
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| CN106546754A (en) * | 2016-12-09 | 2017-03-29 | 新疆医科大学 | Yimusake table acts on abnormal mucus cross-examination with sexual impotence Syndrome model target point protein and its screening technique |
| CN106645748A (en) * | 2016-12-16 | 2017-05-10 | 新疆医科大学 | Mammalian differential serum protein and differentially expressed protein system of penile smooth muscle tissue of abnormal and phlegmatic type of impotence syndrome and screening method and application |
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