CN103125439A - Method for building animal model with hyperuricemia-combined diabetes - Google Patents
Method for building animal model with hyperuricemia-combined diabetes Download PDFInfo
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Abstract
The invention discloses a method for building an animal model with hyperuricemia-combined diabetes. The method comprises the following steps: 1) feeding an target animal with fodder containing 10% (by mass) of yeast powder; subjecting the target animal to a lavage with a 4% (by mass) adenine water solution according to the following dosage of 100 milligram of adenine per day per kilogram of body weight until the blood uric acid level of the target animal starts falling; 2) halving the dosage of the adenine water solution in step 1), subjecting the target animal to a subcutaneous injection with an oteracil potassium emulsion twice a day according to the following dosage of 100 milligram of oteracil potassium emulsion per day per kilogram of body weight until the target animal shows clinical symptoms of diabetes, and then obtaining the animal model with the hyperuricemia-combined diabetes. The method builds the animal model with the hyperuricemia-combined diabetes for the first time, the animal model has typical disease symptoms, the quantity index has statistical significance, and studies such as glucose clamp test and immunohistochemistry also prove that the modeling is successful. Therefore, the method for building the animal model with the hyperuricemia-combined diabetes is a reliable and practical technique.
Description
Technical field
The present invention relates to a kind of method that builds the animal model of hyperuricemia complication with diabetes.
Background technology
In recent years, along with growth in the living standard, the adjustment of dietary structure, the incidence of disease of high lithemia complication with diabetes rises year by year.Studies show that in a large number, the diabetic is occurred together the ratio of hyperuricemia apparently higher than the non-diabetic person, the people of 20%-50% complication with diabetes simultaneously all in Patients with Hyperuricemia, and the incidence of hyperuricemia is about 35% in Old Type-2 Diabetic Patients.Clinical studies show, independent risk factor as atherosclerotic and coronary heart disease, hyperuricemia also has significant impact to chronic complicating diseases of diabetes: the reports such as BoS, hyperuricemia can be accelerated generation and the development of diabetes B patient diabetic nephropathy, Lehto S etc. research finds in the diabetes B patient, and hyperuricemia is the hazards of fatal and non-fatal cerebrovas-cularaccident especially.The diabetic who merges hyperuricemia, the incidence of the complication such as coronary heart disease, cerebral apoplexy, diabetic nephropathy is significantly higher than does not accompany hyperuricemia person.Therefore, the harm of high lithemia complication with diabetes is serious, directly affects people's health, becomes the major disease of needing solution badly.
There is at present report to utilize high fructose diet to bring out rat triglycerides, uric acid, glycometabolism disorderly alternately.Material therefor is the SD rat, and modeling method is: model group contains the modeling feed of fructose.Wherein, modeling needs 60% fructose, and the model cost is high, and uric acid, blood sugar raise very unobvious, model group in the time of the 57th day (UA, 71.69 μ mol/L; GLU, 2.93mmol/L).Therefore, can not be as the pathogenetic typical animal model of research high lithemia complication with diabetes.
The report diabetes animal model is arranged at present, and modeling method is: male Wistar rat, and fasting 12 hours, tail vein injection STZ55mg/kg(is 5.5ml/kg).STZ can bring out many animals and produce diabetes the selective destruction of the beta Cell of islet of certain kind animal, generally adopts rat and mouse to make animal model.The one-tenth mould rate that has the scholar to report abroad to select the male rat modeling is apparently higher than female rats.Although this model blood sugar obviously raises, uric acid is not high, and in addition, although this model clinical manifestation is similar to diabetes B, the mechanism that causes blood sugar to raise is obviously different from diabetes B, therefore can not be as the pathogenetic model of research diabetes B.
In sum, be badly in need of at present a kind of animal model of hyperuricemia complication with diabetes, the pathogenesis of bringing out diabetes B to be used for the research hyperuricemia.
Summary of the invention
An object of the present invention is to provide a kind of method that builds the animal model of hyperuricemia complication with diabetes.
The method of the animal model of structure hyperuricemia complication with diabetes provided by the present invention specifically can comprise the steps:
(1) phase I modeling: with the purpose animal formula forage feed purpose animal of adding 10% dusty yeast; With the 4% adenine aqueous solution, described purpose animal is carried out gavage simultaneously, the consumption of the described 4% adenine aqueous solution is the described purpose the weight of animals of 100mg adenine/sky/kg; Monitor weekly the serum uric acid level of described purpose animal, until the serum uric acid level of described purpose animal when beginning remarkable decline (comparing significant difference with a front monitoring result), carries out following steps (2);
(2) second stage modeling: with the described purpose animal of purpose animal formula forage feed of described interpolation 10% dusty yeast; With the described 4% adenine aqueous solution, described purpose animal is carried out gavage, the consumption of the described 4% adenine aqueous solution is the described purpose the weight of animals of 50mg adenine/sky/kg; Use simultaneously the described purpose animal of Oteracil Potassium emulsion suspension liquid abdominal part hypodermic, the consumption of described Oteracil Potassium emulsion suspension liquid is the described purpose the weight of animals of 100mg Oteracil Potassium/sky/kg, 2 injections of every natural gift (early 8 and late 8 points); Until after described purpose animal shows diabetes clinical symptoms (the plateau serum uric acid level when simultaneously serum uric acid level is with the phase I modeling is compared without marked change), obtain the animal model of hyperuricemia complication with diabetes;
Above each percentage composition is the quality percentage composition.
Another object of the present invention is to provide a kind of cover medicine of the animal model for building the hyperuricemia complication with diabetes.
The cover medicine of the animal model for building the hyperuricemia complication with diabetes provided by the present invention specifically can comprise cover medicine for described phase I modeling, be used for cover medicine and the medication instruction book of described second stage modeling:
Described cover medicine for the phase I modeling specifically is grouped into by following one-tenth:
(a1) feed: for being added with the purpose animal formula feed of dusty yeast, described dusty yeast accounts for and adds 10% of rear feed gross weight;
(a2) gavage medicine: concentration is 4% the adenine aqueous solution, according to the 100mg adenine/described purpose animal body of sky/kg restatement;
Described cover medicine for the second stage modeling is grouped into by following one-tenth:
(b1) feed: for being added with the purpose animal formula feed of dusty yeast, described dusty yeast accounts for and adds 10% of rear feed gross weight;
(b2) gavage medicine: concentration is the adenine aqueous solution of quality percentage composition 4%, according to the 50mg adenine/described purpose animal body of sky/kg restatement;
(b3) injecting drug use: the Oteracil Potassium emulsion suspension liquid, according to the 100mg Oteracil Potassium/described purpose animal body of sky/kg restatement;
Described medication instruction book: use described cover medicine for the phase I modeling for described purpose animal, until the serum uric acid level of described purpose animal is when beginning to descend, then use described cover medicine for the second stage modeling to described purpose animal, discontinue medication after described purpose animal shows the diabetes clinical symptoms.
In the present invention, described diabetes clinical symptoms is mainly manifested in following several respects: 1) diuresis, drink more, become thin; 2) fasting blood-glucose raises, and FPI reduces; 3) OGTT2h blood sugar raises, and OGTT2h insulin reduces; 4) GIR reduces; 5) pancreas islet out-of-shape, insulin cell obviously reduces, glandular tube cellular swelling on every side, the interval diminishes.Wherein, described many, the described rising step 1)-5), described reduction, described minimizing or described diminishing are all with without the corresponding index of the described purpose animal of modeling as a reference.
In the present invention, described Oteracil Potassium emulsion suspension liquid is specifically mixed by each component of following proportioning: sodium carboxymethylcellulose 8g, Oteracil Potassium 25g, physiological saline 1L.
More concrete, in one embodiment of the invention, described Oteracil Potassium emulsion suspension liquid is prepared as follows and obtains: take the sodium carboxymethylcellulose pulvis, be made into physiological saline the emulsion that concentration is 8g/L; Take again Oteracil Potassium, be made into the sodium carboxymethylcellulose of 8g/L the physiological saline emulsion suspension liquid that the Oteracil Potassium final concentration is 25g/L.
Above all described purpose animal formula feeds are daily normal diet, commercially available acquisition.
In the present invention, described purpose animal can be rat.
In the present invention, described purpose animal is specially the wistar rat, more concrete is 12 age in week the wistar rat.
In the present invention, when the described until serum uric acid level of described purpose animal (wistar rat) begins to descend, during finishing in the 7th week being specially when the described phase I, modeling began.
In the present invention, after described until described purpose animal (wistar rat) shows the diabetes clinical symptoms, after finishing in the 10th week (the 9th week after hyperuricemia modeling success) being specially when the described phase I, modeling began; Further, after finishing the 12nd week being preferably when the described phase I, modeling began (hyperuricemia modeling success after the 11st week), as from as described in after finishing in the 13rd week (the 12nd week after hyperuricemia modeling success) when the phase I, modeling began.
Described cover medicine is also belonging to protection scope of the present invention for the preparation of the application in the product of the animal model that builds the hyperuricemia complication with diabetes.
Even so, the method for the animal model of structure hyperuricemia complication with diabetes provided by the present invention also can be described as the animal model that structure has following proterties simultaneously:
Compare 1 with the described purpose animal without modeling) the blood uric acid rising; 2) diuresis, drink more, become thin; 3) fasting blood-glucose raises, and FPI reduces; 4) OGTT2h blood sugar raises, and OGTT2h insulin reduces; 5) GIR reduces; 6) pancreas islet out-of-shape, insulin cell obviously reduces, glandular tube cellular swelling on every side, the interval diminishes.
The present invention has set up the Wistar rat animal model of hyperuricemia complication with diabetes first.This model has typical disease and characterizes, and quantitative index has statistical significance, and the researchs such as anatomic tissue, glucose clamp experiment, SABC also confirm the modeling success.The method can be set up the stable hyperuricemia complication with diabetes rat model that continues, and is a kind of reliable, practical technology.
Description of drawings
Fig. 1 is rat serum uric acid level changing trend diagram.Wherein, * represents P＜0.05 (with the contrast of A group), has significant difference; ☆ represents P＜0.05 (comparing with the D group), has significant difference." which week " of abscissa is for counting when beginning modeling.
Fig. 2 is rat fasting blood-glucose value changing trend diagram.Wherein, * represents P＜0.05 (with the contrast of A group), has significant difference; ☆ represents P＜0.05 (comparing with the D group), has significant difference." which week " of abscissa is for counting when the hyperuricemia modeling is successful.
Fig. 3 is rat limosis insulin value changing trend diagram.Wherein, * represents P＜0.05 (with the contrast of A group), has significant difference." which week " of abscissa is for counting when the hyperuricemia modeling is successful.
Fig. 4 is rat OGTT2h blood glucose value changing trend diagram.Wherein, * represents P＜0.05 (with the contrast of A group), has significant difference; ☆ represents P＜0.05 (comparing with the D group), has significant difference." which week " of abscissa is for counting when the hyperuricemia modeling is successful.
Fig. 5 is rat OGTT2h insulin value changing trend diagram.Wherein, * represents P＜0.05 (with the contrast of A group), has significant difference." which week " of abscissa is for counting when the hyperuricemia modeling is successful.
Fig. 6 is the HE coloration result (amplifying 200 times) of rat (after the modeling success) pancreas during the 10th week.Wherein, A is control group; B is model group.
Fig. 7 is the ImmunohistochemistryResults Results of rat (after the modeling success) pancreas during the 10th week.Wherein, A is the direct collection of illustrative plates (200 *) that SABC obtains, and A-1 is control group, and A-2 is model group; B is to respectively organizing the statistics of the hypocellular severity of rat insulin in A.
Fig. 8 is the ImmunohistochemistryResults Results of rat (after the modeling success) kidney during the 10th week.Wherein, A is the direct collection of illustrative plates (400 *) that SABC obtains, and A-1 is model group, and A-2 is control group; B is to respectively organizing the statistics of the severity of Renal Glomeruli In Rats form in A.
Embodiment
The experimental technique that uses in following embodiment is conventional method if no special instructions.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
1, laboratory animal and feed
Wistar rat (12 week age): Qingdao of Shandong province medicine inspecting institute provides.
The rat pellet: magnificent Fukang, Beijing biotech inc product, catalog number is 1022.
Yeast feed preparation: dusty yeast (UNIPATH company) is evenly admixed in the rat pellet of pulverizing pelletizing of raw material again, and controlling the quality percentage composition of dusty yeast in feed is 10%.
2, various solution
Glucose solution: mass fraction is 30% D/W.
4% adenine solution: mass fraction is 4% the adenine aqueous solution.
Oteracil Potassium solution: electronic balance accurately takes sodium carboxymethylcellulose pulvis (Chemical Reagent Co., Ltd., Sinopharm Group, catalog number 30036328), is made into physiological saline the emulsion that concentration is 8g/L; Take again Oteracil Potassium (a kind of uricase inhibitor), be made into the physiological saline emulsion suspension liquid of Oteracil Potassium final concentration 25g/L with the 8g/L sodium carboxymethylcellulose, put 4 ℃ of refrigerators standby.
3, kit and antibody
(1) insulin radioimmunoassay kit: Beijing North biotechnology research institute.
(2) SABC related reagent:
DAB colour reagent box: company of China fir Golden Bridge in Beijing.
Primary antibodie (rabbit the Chinese People's Anti-Japanese Military and Political College mouse insulin antibody): Cell signal company, 1:400 is made into working solution by volume.
Two anti-(goat anti-rabbit igg, PV-6001): company of China fir Golden Bridge in Beijing.
10% normal goats serum: company of China fir Golden Bridge in Beijing.
4, instrument and equipment
The special-purpose metabolic cage of rat glucose clamp, BIOSEN C-LINE glucose/lactic acid analyzer, WATSON-MARLOW imported with original packaging peristaltic pump and peristaltic pump pump line, pipe before Harvard micro-injection pump and micro pump, blood glucose meter, constant water bath box, low temperature refrigerator, ultra low temperature freezer, vortex mixer, electronic weighing instrument, electronic timer, shadowless lamp, low temperature desk centrifuge, Sysmex CHEMIX2180 type automatic clinical chemistry analyzer.
Experimental data in following embodiment adopts mean ± standard error, and (methods such as t-test are relatively selected in mean ± sem) expression in twos between group, use SPSS13.0 software and carry out statistical analysis.P＜0.05 o'clock is significant difference, P〉0.05 be without significant difference.
The structure of the Wistar rat animal model of embodiment 1, hyperuricemia complication with diabetes
One, the experiment grouping is with diet program
70 wistar rats (12 age in week) are divided into 2 groups at random:
A group: control group (30)
B group: model group (40)
model group (B group) rat all raises to contain the yeast feed of 10% dusty yeast, and with the consumption gavage of 4% adenine solution according to 100mg adenine/(kg body weight d), when monitoring rat serum uric acid level begins to descend (when the 7th week finished from the beginning modeling), improve modeling method, model group (B group) raises to contain the yeast feed of 10% dusty yeast, and with the consumption gavage of 4% adenine solution according to 50mg adenine/(kg body weight d), use simultaneously the Oteracil Potassium emulsion suspension liquid to carry out abdominal part hypodermic according to the consumption of 100mg Oteracil Potassium/(kg body weight d), inject at twice (early and late 8 points) every day at 8, to keep rat hyperuricemia state, control group (A group) is all raised with common rat pellet, and gives with volume distilled water gavage.
In the 12nd week from the beginning modeling (the 11st week after hyperuricemia modeling success), every physiological and biochemical index of model group rat changes after high purine diet and hypodermic injection uric acid enzyme inhibitor for observation stops, B is organized rat is divided into again 2 groups, obtain following three groups:
C group: control group (10)---derive from the A group;
D group: model recovery group (7)---derive from the B group;
E group: continue model group (8)---derive from the B group;
Continue model group (E group) and continue to give high purine diet (diet style is organized with above-mentioned B) and Oteracil Potassium emulsion suspension liquid abdominal part hypodermic; Model recovery group (D group) and control group (C group) are all raised with common rat pellet, and give with volume distilled water gavage.
During this time, foot couple hyperuricemia and the diabetes index of correlation of respectively organizing rat detects weekly, to determine whether model successfully constructs.
Two, the detection of hyperuricemia and diabetes index of correlation
1, the detection of hyperuricemia related biochemical indicator
Before measuring index of correlation, each organizes Rat Septal curfew food 14 hours, then angular vein blood sampling, isolate serum, measure blood uric acid (UA), creatinine (CRE), urea nitrogen (BUN), triglycerides (TG), T-CHOL (TC) on Sysmex CHEMIX2180 type automatic clinical chemistry analyzer.
The measurement result of blood uric acid (UA) as shown in Figure 1, as can be seen from the figure, the model group rat (namely counted for 2 weeks rear) at high purine diet after 2 weeks when beginning modeling serum uric acid level obviously raises, and comparing difference with control group all has significance (p＜0.05), judges hyperuricemia modeling success.Count in the time of will certainly beginning modeling and be designated as rear the 1st week of hyperuricemia modeling success the 2nd week.Until when counting the 7th week (the 6th week after hyperuricemia modeling success) when certainly beginning modeling, the serum uric acid level of model group rat begins remarkable decline (count the 6th all monitoring results when certainly beginning modeling and compare significant difference), this may be relevant with uricase in hyperuricemia activates the rat body, therefore in above step 1, in order to continue to keep the hyperuricemia state of rat, the diet program of model group improves.After improvement, the model group hyperuricemia can be kept and surpass 14 weeks (seeing the lasting model group shown in the E group in Fig. 1), and before and after improving, model group rat serum uric acid does not have significant change, and this shows that hyperuricemia model is stable.And model recovery group (D group) is from recovering full diet (counting for the 12nd week when certainly beginning modeling), and its serum uric acid level begins remarkable decline.
The measurement result of creatinine (CRE), urea nitrogen (BUN), triglycerides (TG) and T-CHOL (TC) as shown in Table 1 and Table 2.Hyperuricemia modeling success is during rear the 1st week, and monitoring model group rat TC, BUN, CRE and control group comparing difference have statistical significance, the TG no significant difference; Again monitor These parameters after hyperuricemia modeling success during the 8th week, difference is identical during still with the 1st week.
The 1st all blood biochemistry of rats index determining results after table 1 hyperuricemia modeling success
*: P＜0.05 (comparing with control group) has significant difference.
The 8th all blood biochemistry of rats index determining results after table 2 hyperuricemia modeling success
*: P＜0.05 (comparing with control group) has significant difference.
2, the detection of diabetes relevant clinical and biochemical indicator
(1) observation of clinical symptoms
With the carrying out of experiment, the 8th all pattern drawing groups (or lasting model group) rat is compared with control group after hyperuricemia modeling success, shows obvious typical diabetes clinical symptoms: diuresis, drink more, become thin.
(2) mensuration of fasting blood-glucose and insulin
Before measuring index of correlation, each organizes Rat Septal curfew food 14 hours.Then, on the one hand, to cut tail method blood sampling 5-10 μ l, blood glucose meter is measured fasting blood sugar.On the other hand, the angular vein blood sampling after separation of serum, is measured the FPI value with the insulin radioimmunoassay kit, and concrete operations are carried out referring to the kit specification.Choose at random 10 rats for every group and test, result is got the mean value of 10 rats.
The measurement result of fasting blood-glucose as shown in Figure 2, as can be seen from the figure, carrying out with experiment, the 9th all pattern drawing groups (or lasting model group) rat fasting blood-glucose is compared with control group after hyperuricemia modeling success, begin to show difference, statistical significance (p＜0.05) is arranged, and model group (or lasting model group) rat fasting blood-glucose value totally is rising situation, until the 14th week after the modeling that last detects success.And model recovery group (D group) (the 12nd week from beginning modeling, i.e. the 11st week after hyperuricemia modeling success) from the recovery full diet, its fasting blood glucose level begins remarkable decline.
The measurement result of FPI as shown in Figure 3, as can be seen from the figure, test early stage model group rat limosis insulin i.e. variant with control group (p＜0.05), this may affect insulin secretion with hyperuricemia, and causing in a short time, insulin level reduces relevant.
(3) mensuration of OGTT2h blood sugar and OGTT2h insulin
Oral glucose tolerance test (OGTT): according to the consumption of 2g/kg body weight, each is organized rat oral gavage with 30% glucose solution.After glucose load 2 hours.On the one hand, to cut tail method blood sampling 5-10 μ l, blood glucose meter is measured the OGTT2h blood glucose value.On the other hand, the angular vein blood sampling after separation of serum, is measured OGTT2h insulin value with the insulin radioimmunoassay kit, and concrete operations are carried out referring to the kit specification.Choose at random 10 rats for every group and test, result is got the mean value of 10 rats.
The measurement result of OGTT2h blood sugar as shown in Figure 4, as can be seen from the figure, carrying out with experiment, the 8th all pattern drawing groups (or lasting model group) rat OGTT2h blood sugar is compared with control group after hyperuricemia modeling success, begin to show difference, statistical significance (p＜0.05) is arranged, and model group (or lasting model group) rat OGTT2h blood glucose value totally is rising situation, until the 14th week after hyperuricemia modeling success that last detects.And model recovery group (D group) (the 12nd week from beginning modeling, i.e. the 11st week after hyperuricemia modeling success) from the recovery full diet, its OGTT2h blood sugar level begins remarkable decline.
The measurement result of OGTT2h insulin as shown in Figure 5, as can be seen from the figure, test early stage model group rat OGTT2h insulin i.e. variant with control group (p＜0.05), this may affect insulin secretion with hyperuricemia, and causing in a short time, insulin level reduces relevant.
(4) GIR (GIR) is measured
Respectively at from the beginning modeling the 2nd, 6,10,13 weeks 5 rats of random choose from every group carry out hyperinsulinism-positive glucose clamp experiment, calculate GIR (GIR), rat insulin susceptibility is respectively organized in representative.Hyperinsulinism-positive glucose clamp experiment is specifically carried out as follows: make a 1cm left and right otch after rat anesthesia on the neck median line, separate jugular vein and arteria carotis, insert respectively silicone tube and fixing, skin closure; Approximately after 5-7 days, under the empty stomach state through jugular vein and arteria carotis respectively after external silicone tube, input respectively insulin and glucose, the infusion of insulin constant rate, adjust glucose infusion speed, to make glycemic control 5mmol/L (about 4.5～5.5mmol/L), after the glucose input begins 60min, when continuous 6 blood glucose values are arranged all in above-mentioned scope, namely think to reach stable state.Then according to glucose infusion amount and the time of stable state time period, calculate glucose input rate (GIR), as the index of weighing animal insulin susceptibility.
Result is as shown in table 3, from the 2nd week the beginning modeling, model group rat GIR is compared with control group namely obvious significant difference (P＜0.05), and the insulin sensitivity of model group is significantly lower than control group, As time goes on, this species diversity sustainable existence (P＜0.05).In addition, when the 13rd week from the beginning modeling, model recovery group (D group) is compared with lasting model group (E group), and its insulin sensitivity shows rise situation.
Table 3 is respectively organized GIR (GIR) the measurement result unit of rat: (mg/kg.min)
*: P＜0.05 (with the contrast of A group) has significant difference;
☆: P＜0.05 (with the contrast of C group) has significant difference.
3, respectively organize the tissue specimen analysis of rat kidney and pancreas
Experimental result based on above each several part detection, be chosen in the 10th week (model group blood sugar level rising after hyperuricemia modeling success, compared notable difference with control group) time, to put to death after the part rat fasting 12h in each group, get its pancreas and kidney, with 10% neutral formalin fixed preparation, paraffin embedding.All samples carry out HE dyeing and SABC (the SABC related reagent is seen the above part of embodiment embodiment).
The HE coloration result of pancreas as shown in Figure 6, as can be seen from the figure, the model group rat is compared the pancreas islet shape and becomes irregular with control group during the 10th week after hyperuricemia modeling success, glandular tube cellular swelling on every side, the interval diminishes.The ImmunohistochemistryResults Results of pancreas as shown in Figure 7, as can be seen from the figure, the model group rat is compared the pancreas islet shape and becomes irregular with control group when hyperuricemia modeling success the rear the 10th is all, insulin cell obviously reduces, compare with Normal group, model group rat insulin cell dyeing area obviously reduces, glandular tube cellular swelling on every side, and the interval diminishes.All these pathological characteristics are consistent with clinical diabetes.
The ImmunohistochemistryResults Results of kidney as shown in Figure 8, as can be seen from the figure, the atrophy of model group glomerulus, quantity reduces (shown in long arrow); The renal tubule cloudy swelling, between renal tubule reaches, there is more urate crystal deposition (shown in short arrow) at the matter position.It is consistent that all these pathological characteristics and clinical hyperuricemia are brought out gouty nephropathy.
Comprehensive above experimental result, the method for summing up the wistar rat model that builds the hyperuricemia complication with diabetes is as follows:
(1) phase I modeling: with adding 10%(quality percentage composition) the forage feed wistar rat of dusty yeast; Use simultaneously 4%(quality percentage composition) the adenine aqueous solution carries out gavage to described wistar rat, and the consumption of the described 4% adenine aqueous solution is the described wistar rat body weight of 100mg adenine/sky/kg; Monitor weekly the serum uric acid level of described wistar rat, until the serum uric acid level of described purpose animal when beginning remarkable decline (the 7th week finished from the beginning modeling), carries out following steps (2);
(2) second stage modeling: with the described wistar rat of the forage feed of described interpolation 10% dusty yeast; With the described 4% adenine aqueous solution, described wistar rat is carried out gavage, the consumption of the described 4% adenine aqueous solution is the described wistar rat body weight of 50mg adenine/sky/kg; Use simultaneously the described wistar rat of Oteracil Potassium emulsion suspension liquid abdominal part hypodermic, the consumption of described Oteracil Potassium emulsion suspension liquid is the described wistar rat body weight of 100mg Oteracil Potassium/sky/kg, 2 injections of every natural gift; Until described wistar rat shows diabetes clinical symptoms (the 9th week after hyperuricemia modeling success, preferably from rear the 11st week of hyperuricemia modeling success, as the 12nd week after the hyperuricemia modeling is successful) after, the animal model of acquisition hyperuricemia complication with diabetes.
Claims (9)
1. a method that builds the animal model of hyperuricemia complication with diabetes, comprise the steps:
(1) phase I modeling: with the purpose animal formula forage feed purpose animal of adding 10% dusty yeast; With the 4% adenine aqueous solution, described purpose animal is carried out gavage simultaneously, the consumption of the described 4% adenine aqueous solution is the described purpose the weight of animals of 100mg adenine/sky/kg; Until the serum uric acid level of described purpose animal when beginning to descend, carries out following steps (2);
(2) second stage modeling: with the described purpose animal of purpose animal formula forage feed of described interpolation 10% dusty yeast; With the described 4% adenine aqueous solution, described purpose animal is carried out gavage, the consumption of the described 4% adenine aqueous solution is the described purpose the weight of animals of 50mg adenine/sky/kg; Use simultaneously the described purpose animal of Oteracil Potassium emulsion suspension liquid hypodermic injection, the consumption of described Oteracil Potassium emulsion suspension liquid is the described purpose the weight of animals of 100mg Oteracil Potassium/sky/kg, 2 injections of every natural gift; Until after described purpose animal shows the diabetes clinical symptoms, obtain the animal model of hyperuricemia complication with diabetes;
Above each percentage composition is the quality percentage composition.
2. be used for to build the cover medicine of the animal model of hyperuricemia complication with diabetes, comprise cover medicine for the phase I modeling, be used for cover medicine and the medication instruction book of second stage modeling:
Described cover medicine for the phase I modeling is grouped into by following one-tenth:
(a1) feed: for being added with the purpose animal formula feed of dusty yeast, described dusty yeast accounts for and adds 10% of rear feed gross weight;
(a2) gavage medicine: concentration is the adenine aqueous solution of quality percentage composition 4%, according to the 100mg adenine/described purpose animal body of sky/kg restatement;
Described cover medicine for the second stage modeling is grouped into by following one-tenth:
(b1) feed: for being added with the purpose animal formula feed of dusty yeast, described dusty yeast accounts for and adds 10% of rear feed gross weight;
(b2) gavage medicine: concentration is 4% the adenine aqueous solution, according to the 50mg adenine/described purpose animal body of sky/kg restatement;
(b3) injecting drug use: the Oteracil Potassium emulsion suspension liquid, according to the 100mg Oteracil Potassium/described purpose animal body of sky/kg restatement;
Described medication instruction book: use described cover medicine for the phase I modeling for described purpose animal, until the serum uric acid level of described purpose animal is when beginning to descend, then use described cover medicine for the second stage modeling to described purpose animal, discontinue medication after described purpose animal shows the diabetes clinical symptoms.
3. method according to claim 1, or cover medicine claimed in claim 2 is characterized in that: described Oteracil Potassium emulsion suspension liquid is mixed by each component of following proportioning: sodium carboxymethylcellulose 8g, Oteracil Potassium 25g, physiological saline 1L.
4. arbitrary described method or cover medicine according to claim 1-3, it is characterized in that: described purpose animal is rat.
Method according to claim 4 or the cover medicine, it is characterized in that: described rat is the wistar rat.
6. method according to claim 5 or cover medicine is characterized in that: when the described until serum uric acid level of described purpose animal begins to descend, during for finishing in the 7th week when the described phase I, modeling began.
7. according to claim 5 or 6 described methods or cover medicine is characterized in that: after described until described purpose animal shows the diabetes clinical symptoms, for after finishing in the 10th week when the described phase I, modeling began.
8. method according to claim 7 or cover medicine is characterized in that: after described until described purpose animal shows the diabetes clinical symptoms, for after finishing in the 12nd week when the described phase I, modeling began.
In claim 2-8 arbitrary described cover medicine for the preparation of the application in the product of the animal model that builds the hyperuricemia complication with diabetes.
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| CN105850869A (en) * | 2016-03-31 | 2016-08-17 | 青岛大学附属医院 | Method and device for constructing animal model suffering from diabetes induced by high uric acid content |
| CN106900652A (en) * | 2017-03-16 | 2017-06-30 | 青岛大学 | A kind of construction method of hyperuricemia model |
| CN108524553A (en) * | 2018-07-09 | 2018-09-14 | 武汉轻工大学 | A kind of construction method of subacute hyperuricemia renal damage mouse model |
| CN110338139A (en) * | 2019-07-03 | 2019-10-18 | 安徽省立医院 | A kind of construction method of gout animal model and application |
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| CN111939161A (en) * | 2013-09-26 | 2020-11-17 | 华安医学股份有限公司 | AMPK-activating compounds and uses thereof |
| CN113040093A (en) * | 2021-03-11 | 2021-06-29 | 四川大学华西医院 | Application of oteracil potassium and adenine in inducing rat hyperuricemia renal injury model |
| CN115039737A (en) * | 2021-08-09 | 2022-09-13 | 北京中医药大学 | Method for establishing urate nephrosis animal model |
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| CN111939161A (en) * | 2013-09-26 | 2020-11-17 | 华安医学股份有限公司 | AMPK-activating compounds and uses thereof |
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| WO2015170694A1 (en) * | 2014-05-08 | 2015-11-12 | 株式会社フェニックスバイオ | Hyperuricemia model |
| CN105850869A (en) * | 2016-03-31 | 2016-08-17 | 青岛大学附属医院 | Method and device for constructing animal model suffering from diabetes induced by high uric acid content |
| CN106900652A (en) * | 2017-03-16 | 2017-06-30 | 青岛大学 | A kind of construction method of hyperuricemia model |
| CN108524553A (en) * | 2018-07-09 | 2018-09-14 | 武汉轻工大学 | A kind of construction method of subacute hyperuricemia renal damage mouse model |
| CN110338139A (en) * | 2019-07-03 | 2019-10-18 | 安徽省立医院 | A kind of construction method of gout animal model and application |
| CN110338139B (en) * | 2019-07-03 | 2021-10-26 | 安徽省立医院 | Construction method and application of gout animal model |
| CN111887202A (en) * | 2020-08-26 | 2020-11-06 | 安发（福建）生物科技有限公司 | Method for constructing mouse model of acute hyperuricemia |
| CN111772050A (en) * | 2020-08-28 | 2020-10-16 | 常州鼠一鼠二生物科技有限公司 | Rat hyperuricemia model feed and preparation method and application thereof |
| CN113040093A (en) * | 2021-03-11 | 2021-06-29 | 四川大学华西医院 | Application of oteracil potassium and adenine in inducing rat hyperuricemia renal injury model |
| CN115039737A (en) * | 2021-08-09 | 2022-09-13 | 北京中医药大学 | Method for establishing urate nephrosis animal model |
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