CN111557273A - Method for inducing type 2 diabetes animal model by low temperature and diet rhythm regulation - Google Patents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A—HUMAN NECESSITIES
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- A01K2267/0362—Animal model for lipid/glucose metabolism, e.g. obesity, type-2 diabetes
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Abstract
The invention discloses a method for regulating and inducing a type 2 diabetes animal model by low temperature and diet rhythm, which induces animals to form the type 2 diabetes animal model with weight gain, hyperlipidemia, hyperglycemia, hyperinsulinemia and insulin resistance as main marks through the regulation and control of low temperature environment in combination with the change of the diet rhythm of feed or independently through the change of the diet rhythm of feed. The experimental research results show that the low-temperature environment and the night time-limited diet can induce SD rats to form the main signs of type 2 diabetes such as weight gain, hyperlipidemia, hyperglycemia, hyperinsulinemia, insulin resistance and the like, can obviously reduce the diversity of intestinal microorganisms, is close to the pathogenesis mode of human type 2 diabetes, and the constructed diabetes animal model can be used for researching the pathogenesis of type 2 diabetes, searching relevant targets for early diagnosis of the type 2 diabetes animal model, and researching and developing therapeutic drugs and prevention and treatment methods.
Description
Technical Field
The invention relates to a construction method of a type 2 diabetes animal model, in particular to a method for quickly inducing the type 2 diabetes animal model through low-temperature environment and circadian rhythm regulation, belonging to the technical field of biological medicine.
Background
Diabetes has become one of the most serious public health problems worldwide in the 21 st century, most of which are type 2 diabetes. Type 2 diabetes, also known as non-insulin dependent diabetes mellitus, is an endocrine-metabolic disease with hyperglycemia as a main marker, which is caused by relatively insufficient insulin secretion or reduced sensitivity of target cells to insulin, resulting in metabolic disorders of sugar, protein, fat and secondary water and electrolyte. The clinical manifestations are "three more and one less", "three more" refers to polyuria, polydipsia and polyphagia, and "one less" refers to emaciation with symptoms of fatigue, polydipsia and the like. Researches show that the high incidence of the type 2 diabetes in the society at present is closely related to the improvement of the living standard of human beings and the change of life style and behaviors (particularly diet in day and night), so that the analysis of the incidence reason of the type 2 diabetes has important significance for the early diagnosis and the prevention and the treatment of the type 2 diabetes.
The current type 2 diabetes model is mainly composed of an animal model of spontaneous type 2 diabetes and an animal model of experimental type 2 diabetes, wherein the application and popularization of the spontaneous type 2 diabetes in scientific research are limited due to the defects of relatively few sources, high requirements on feeding and breeding conditions, expensive animals and the like of the spontaneous type 2 diabetes. The experimental type 2 diabetes animal model is characterized in that the pancreas or islet beta cells are damaged by methods such as high-fat diet, Streptozocin (STZ), alloxan and the like to cause insulin deficiency, or various antagonists are used for resisting the action of insulin to cause experimental diabetes or experimental hyperglycemia. In addition, the early pathophysiological basis of both spontaneous hypertensive rats and type 2 diabetic patients is insulin resistance, and therefore, the early pathophysiological basis plays an important role in the research of diabetes mechanism and the evaluation of drug efficacy. However, the methods for making these animal models are significantly different from the onset of type 2 diabetes in humans, and in particular, the role of lifestyle and behavioral changes in the course of type 2 diabetes cannot be revealed.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for inducing an animal model of type 2 diabetes by regulating and controlling low temperature and diet rhythm, researching the pathogenesis of the animal model of type 2 diabetes by simulating life style and behavior change, and providing a new way for early diagnosis and prevention of type 2 diabetes.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a method for regulating and inducing animal model of type 2 diabetes by low temperature and diet rhythm includes such steps as regulating and controlling low temperature environment, feeding feed, and changing the diet rhythm of feed or only feeding feed to induce animal to become animal model of type 2 diabetes with the symptoms of weight increase, hyperlipemia, hyperglycemia, hyperinsulinemia and insulin resistance.
The method for regulating and inducing the animal model of type 2 diabetes by the low temperature and the dietary rhythm is characterized in that the low temperature environment is lower than the environment temperature suitable for animals, and the time period and the time duration of the time-limited feeding are changed after the dietary rhythm is changed to the time-limited feeding for a period of time.
As a preferred scheme, the method for regulating and inducing the animal model of type 2 diabetes at low temperature and dietary rhythm is used, and the low-temperature environment is 16-25 ℃.
The method for inducing the animal model of type 2 diabetes by low temperature and diet rhythm regulation is preferably implemented by feeding a high-fat feed or a common feed.
In the method for inducing the animal model of type 2 diabetes by low temperature and regulation of food rhythm, the animal is preferably rat, mouse, nude mouse, rabbit, guinea pig, monkey or chimpanzee.
As a preferable scheme, the method for inducing the type 2 diabetes animal model by low temperature and diet rhythm regulation has the induction time of 10-20 days.
Application of low temperature and diet rhythm regulation and control method for inducing type 2 diabetes animal model in diabetes treatment.
The application can be used for developing the reagent and the method for early diagnosis of diabetes by using the diabetes animal model constructed by the method.
The application can also be used for discovering new treatment targets and developing diabetes treatment medicines by using the diabetes animal model constructed by the method.
The application can also use the diabetes animal model constructed by the method to develop other treatment methods for diabetes.
The invention has the beneficial effects that: in experimental research, the applicant finds that the change of low-temperature environment and day-night dietary rhythm can induce the main signs of type 2 diabetes such as rat weight gain, hyperlipidemia, hyperglycemia, hyperinsulinemia and insulin resistance in about 10 days. According to the experiment of the influence of the environmental temperature and the circadian rhythm on the weight and the glycolipid metabolic components of the rat and the experiment of the influence of the low-temperature environment and the circadian rhythm on the intestinal microorganism diversity of the rat, the invention proves that the change of the low-temperature environment and the circadian rhythm can induce the glycolipid metabolic disorder of the rat and reduce the intestinal microorganism abundance of the rat, so that the early symptoms of the type 2 diabetes mellitus are met, and a new way and a new thought are provided for researching the pathogenesis of the type 2 diabetes mellitus, searching the related target spot of early diagnosis and formulating a prevention and treatment method.
The invention has the advantages that: (1) the invention can induce early symptoms of type 2 diabetes in a short time (10 days). (2) The construction method of the invention is simple, and is easy to operate only by the change of low-temperature environment and day-night dietary rhythm. (3) The 2-type diabetes animal model constructed by the invention is more in line with the current pathogenesis mode of human 2-type diabetes, and can be used for researching the pathogenesis of the 2-type diabetes, searching relevant targets for early diagnosis, and researching and developing a treatment medicament and a prevention and treatment method for the diabetes.
Drawings
FIG. 1 is a schematic diagram showing the effects of the low temperature environment and the circadian rhythm of the present invention on the body weight and glycolipid metabolic components of rats;
FIG. 2 is a schematic diagram showing the effect of the low temperature environment and the circadian rhythm of the present invention on rat insulin and insulin resistance coefficient (IR);
FIG. 3 is a schematic illustration of the effect of the low temperature environment and circadian rhythm of the invention on the abundance of intestinal microbes in rats;
FIG. 4 is a schematic diagram showing the effect of the low temperature environment and the circadian rhythm of the present invention on rat intestinal portal microorganisms.
The present invention will be further described with reference to the following detailed description and accompanying drawings.
Detailed Description
Example 1: the effects of changes in ambient temperature and circadian rhythm on rat body weight and glycolipid metabolism.
1.2 Induction of animal models of diabetes mellitus
150g-200g male SD rats with 4 weeks of age are selected. The natural light is irradiated for 12 hours every day, and the adaptive feeding is carried out for 1 week. Through different temperatures (30 ℃ at high temperature, 25 ℃ at proper temperature and 16 ℃ at low temperature), time-limited diet (day and night) and high-fat diet influence the diet rhythm and structure of rats, a high-temperature normal feed day-time diet control group, a high-temperature normal feed night diet control group, a proper-temperature normal feed day-time diet control group, a proper-temperature normal feed night diet control group, a low-temperature normal feed day-time diet control group, a low-temperature normal feed night diet control group, a high-fat diet day-time diet control group and a high-fat diet night diet control group are arranged for 8 groups, the change of physiological indexes of rats such as food intake, weight and the like is observed, and a full-automatic biochemical analyzer is adopted to detect the indexes such as glucose, triglyceride, total cholesterol and the like in serum.
The results show that the food intake of the rats tends to be stable in the nighttime and daytime under the condition of the moderate temperature (see figure 1, P is more than 0.01 and less than or equal to 0.05, and P is more than or equal to 0.001 and less than or equal to 0.01), the food intake at night under the high-temperature and low-temperature feeding environments is obviously increased, the weight of the rats in the high-temperature daytime feeding limit group is obviously lower than that of the moderate temperature group, and the weight of the rats in the low-temperature night feeding limit group is obviously higher than that of the.
2. Analysis of physiological index and glycolipid metabolic component
The influence of diet circadian rhythm change and environmental temperature on blood sugar and blood fat is analyzed (see figure 2, P is more than 0.01 and less than or equal to 0.05, P is more than 0.001 and less than or equal to 0.01), and the blood sugar of the rats in the night feeding-restricted group is obviously increased along with the reduction of the environmental temperature (P is less than or equal to 0.01); the total cholesterol and the high-density lipoprotein in the serum of the rats in the day-time feeding control group and the night-time feeding control group are increased along with the environmental temperature; the triglyceride of the rat fed with the low-temperature night time-limited diet is obviously higher than that of other groups (P is less than or equal to 0.05), but has no obvious correlation with the environmental temperature; low density lipoprotein in rat serum has some correlation with ambient temperature, but the results are not significant. It is suggested that the normal metabolism of sugar in the body is easily caused by eating at night, and the disturbance of glycolipid metabolism is easily caused by the change of low-temperature environment.
Example 2: the influence of low temperature environment and circadian rhythm on the intestinal microbial diversity of rats.
Analyzing the influence of low-temperature environment and day-night diet rhythm on the diversity of rat intestinal microorganisms by using a microorganism diversity sequencing technology, and displaying that the low-temperature environment can obviously reduce the abundance of rat intestinal microorganisms, wherein the low-temperature night feeding-limited group rat microorganisms have the lowest abundance (see figure 3, 0.01< P < 0.05 and 0.001< P < 0.01), the night feeding-limited diet can obviously reduce bacteroides (bacteroides) and actinomycetes (actinomycetes) in the intestinal microorganisms, the Firmicutes (Firmicutes) and Proteobacteria (Proteobacteria) are increased, the abundance of the Firmicutes in the low-temperature night feeding-limited group is far higher than that of other groups (see figure 4, the Y axis represents species name under a certain taxonomic level, the X axis represents average relative abundance in different groups of species, columns of different numbers represent different groups, the rightmost side represents a P value, 0.01< P < 0.05 and P < 0.01), suggesting that low temperature environments may promote colonization by firmicutes microorganisms.
As a result: the low-temperature environment and the night time-limited diet can induce SD rats to form main signs of type 2 diabetes such as weight gain, hyperlipidemia, hyperglycemia and the like, and the low-temperature environment and the night diet can obviously reduce the diversity of intestinal microorganisms and are close to the morbidity mode of the type 2 diabetes of human beings.
The embodiments of the present invention are not limited to the above-described examples, and various changes made without departing from the spirit of the present invention are within the scope of the present invention.
Claims (10)
1. A method for inducing a type 2 diabetes animal model by low temperature and diet rhythm regulation is characterized in that: the method combines the regulation and control of low temperature environment with the change of the feeding rhythm of the feed, or independently induces animals to form a type 2 diabetes animal model with weight gain, hyperlipemia, hyperglycemia, hyperinsulinemia and insulin resistance as main markers.
2. The method of claim 1 for inducing an animal model of type 2 diabetes by low temperature and regulation of dietary rhythm, wherein: the low temperature environment is lower than the environment temperature suitable for animals, and the time period and the time length of the time-limited feeding are changed after the food rhythm is changed to the time-limited feeding for a period of time.
3. The method of claim 2 for inducing an animal model of type 2 diabetes by low temperature and regulation of dietary rhythm, wherein: the low-temperature environment is 16-25 ℃.
4. The method of claim 1 for inducing an animal model of type 2 diabetes by low temperature and regulation of dietary rhythm, wherein: the feed for feeding adopts high-fat feed or common feed.
5. The method of claim 1 for inducing an animal model of type 2 diabetes by low temperature and regulation of dietary rhythm, wherein: the animal is a rat, mouse, nude mouse, rabbit, guinea pig, monkey or chimpanzee.
6. The method of claim 1 for inducing an animal model of type 2 diabetes by low temperature and regulation of dietary rhythm, wherein: the induction time is 10-20 days.
7. Use of the method of inducing an animal model of type 2 diabetes mellitus at low temperatures and with regulation of dietary rhythm according to any one of claims 1 to 6 in the treatment of diabetes.
8. Use according to claim 7, characterized in that: the diabetes animal model constructed by the method is used for developing early diagnosis reagents and methods for diabetes.
9. Use according to claim 7, characterized in that: the diabetes animal model constructed by the method is used for discovering new treatment targets and developing diabetes treatment medicines.
10. Use according to claim 7, characterized in that: the diabetes animal model constructed by the method is used for developing other treatment methods aiming at diabetes.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112088837A (en) * | 2020-09-22 | 2020-12-18 | 西北民族大学 | Construction method of non-alcoholic fatty liver mouse model |
| CN114225012A (en) * | 2021-12-16 | 2022-03-25 | 浙江湃肽生物有限公司 | Method for measuring bioactivity of somaglutide |
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| CN112088837A (en) * | 2020-09-22 | 2020-12-18 | 西北民族大学 | Construction method of non-alcoholic fatty liver mouse model |
| CN114225012A (en) * | 2021-12-16 | 2022-03-25 | 浙江湃肽生物有限公司 | Method for measuring bioactivity of somaglutide |
| CN114225012B (en) * | 2021-12-16 | 2022-10-25 | 浙江湃肽生物股份有限公司 | Method for measuring bioactivity of somaglutide |
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