CN101619304A - Constructing method of ovary insulin resistant model - Google Patents
Constructing method of ovary insulin resistant model Download PDFInfo
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- CN101619304A CN101619304A CN200910162298A CN200910162298A CN101619304A CN 101619304 A CN101619304 A CN 101619304A CN 200910162298 A CN200910162298 A CN 200910162298A CN 200910162298 A CN200910162298 A CN 200910162298A CN 101619304 A CN101619304 A CN 101619304A
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Abstract
The invention discloses a construction method of an ovary insulin resistant model, which belongs to the field of construction of an animal pathologic tissue model. The construction method comprises the following step: after a small mouse is killed, taking out the ovary and culturing the ovary by adopting an in vitro culture medium in a CO2 culture box for in vitro culture, wherein during the in vitro culture, hexadecadrol is added into the in vitro culture medium for induction. The ovary insulin resistant model is constructed by adopting the hexadecadrol to induce and culturing the ovary of the small mouse in vitro, and the change condition of the functions of the ovary in the insulin resistant model is evaluated by detecting the content of glucose in culture liquid. Shown by an experimental result, the effect of inducing ovary insulin to resist by the hexadecadrol has time dose dependency, and after the ovary is treated by the hexadecadrol, the intake of the ovary to the glucose is reduced. Under the stimulation of the insulin, the intake of the ovary treated by the hexadecadrol to the glucose is obviously reduced, thereby indicating that the ovary insulin resistant model is ideal.
Description
Technical field
The present invention relates to a kind of construction process of animal model, relate in particular to a kind of construction process of ovary insulin resistant model, belong to the structure field of animal pathological model.
Background technology
Polycystic ovarian syndrome (PCOS) is the common cause of infertility of reproductive age, serves as main performance unusually with ovarian function, and relevant with the compensatory hyperinsulinemia with insulin resistant.But current research only limits to classical target tissue-skeletal muscle, fat and the liver etc. of insulin action.The acceptor of Regular Insulin also is present in the ovary tissue simultaneously, and this shows that ovary is another important target organ of insulin action.Explain that with the insulin resistant outside the ovary tissue and the hyperinsulinemia of periphery the dysfunction of ovary itself is inapt obviously.Therefore, to study insulin resistant be necessary to the influence of ovarian function to the model that makes up a kind of ovary insulin resistant.
Summary of the invention
The construction process that the purpose of this invention is to provide a kind of new ovary insulin resistant model;
The objective of the invention is to be achieved through the following technical solutions:
A kind of construction process of ovary insulin resistant model may further comprise the steps: mouse is put to death the back and takes out ovary employing in-vitro culture medium at CO
2Carry out vitro culture in the incubator; Wherein, adding dexamethasone when carrying out vitro culture in in-vitro culture medium induces;
Wherein, described in-vitro culture medium is preferably the F-12 substratum, and its moiety comprises: 5%BSA, 0.1%DMSO, 10nmol/l Regular Insulin; Described CO
2CO in the incubator
2Content be 5% (V/V), relative air humidity is 95%;
In order to reach better effect, adding concentration when carrying out vitro culture in in-vitro culture medium is the external evoked ovary 48h-72h of 100-300nmol/L dexamethasone; Preferred, adding concentration when carrying out vitro culture in in-vitro culture medium is the external evoked ovary 48h of dexamethasone of 300nmol/L;
The present invention is by the vitro culture mouse ovarian, and induced by dexamethasone is set up ovary insulin resistant model, by detecting the changing conditions of nutrient solution glucose content assessment insulin resistant model ovarian function.Experimental result shows the effect lifetime dose-dependently of induced by dexamethasone ovary insulin resistant, after Dex handled, ovary descended to the intake of glucose, under insulin stimulating, the ovary that Dex handles is to the more obvious decline of intake of glucose, ovary insulin resistant model ideal.
The present invention adopts different concentration dexamethasone to come the inducing mouse ovary with different induction times, and experimental result is found, uses 300nmol/L induced by dexamethasone 48h, and constructed ovary insulin resistant model is ideal.
The breviary term that arrives involved in the present invention:
Dex: dexamethasone
RT-PCR: reverse transcription polymerase chain reaction
BSA: bovine serum albumin
DMSO: the inferior maple of dimethyl
PCOS: polycystic ovarian syndrome
Description of drawings
The different induced concentration of Fig. 1 dexamethasone and of the influence of different induction times to ovary insulin resistant model; A: base set ovary glucose uptake amount; B: ovary glucose uptake amount behind the insulin stimulating;
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Embodiment
Laboratory animal is provided by Heilongjiang University of Chinese Medicine's Experimental Animal Center, chooses female Kunming white mouse (body weight for (18 ± 1) g/ only), and the mouse that the transvaginal smear is determined cycle stability and is in dioestrus is an experimental subjects.Mouse is got ovary and is used for vitro culture after putting to death.Mouse ovarian takes out the back and peels off surrounding tissue at microscopically, puts into the culture dish of the 50mm that contains physiological saline, uses the mixed solution flushing ovary that is equipped with good DMEM and 5%BSA more than three times.Put into four mouse ovarians at random for every group and carry out vitro culture with the F-12 substratum, the Regular Insulin of nutrient solution composition: 5%BSA, 0.1%DMSO and 10nmol/l, four groups of dexamethasone that add respective concentration wherein are at atmospheric moisture 95%, CO
2Cultivate in the incubator of content 5%.Use the dexamethasone external evoked mouse ovarian 48h of difference and the 72h of different concns (100nmol/L, 300nmol/L), determine the best use of concentration and the time of dexamethasone.Specifically be grouped into: do not use dexamethasone and cultivate ovary 48h group (A1), 100nmol/l induced by dexamethasone 48h group (A2), 300nmol/l induced by dexamethasone 48h group (A3), do not use dexamethasone and cultivate ovary 72h group (B1), 100nmol/l induced by dexamethasone 72h group (B2), 300nmol/l induced by dexamethasone 72h and organize (B3).
The mensuration of nutrient solution glucose content: the mensuration of nutrient solution glucose content is divided into base set and insulin stimulating group.Base set is after application DEX cultivates the corresponding time, to get six groups of nutrient solutions, measures glucose content.The insulin stimulating group is that every group of insulin stimulating 2h that adds 200nmol/L measures the nutrient solution sugar degree again after application DEX cultivates the corresponding time and changes nutrient solution.In base set (Figure 1A), the ovary glucose uptake amount of the induced by dexamethasone of two kinds of concentration all is lower than the identical incubation time group of not using dexamethasone, A3 group nutrient solution glucose uptake amount is starkly lower than A1 group (9.9 ± 0.78vs.36.18 ± 1.7, p＜0.05).Figure 1B is depicted as the variation of respectively organizing ovary glucose uptake amount behind the insulin stimulating.Cultivate 48h and organize the decline that presents the dexamethasone dose-dependently, A3 group glucose uptake amount is starkly lower than A1 group (7.74 ± 0.3vs.38.34 ± 1.2, p＜0.01), prolongs incubation time to 72h, and glucose uptake is not further to descend.These presentation of results are used 300nmol/L induced by dexamethasone 48h, and constructed ovary insulin resistant model is ideal.
Claims (5)
1, a kind of construction process of ovary insulin resistant model may further comprise the steps: mouse is put to death the back and takes out ovary employing in-vitro culture medium at CO
2Carry out vitro culture in the incubator; Wherein, adding dexamethasone when carrying out vitro culture in in-vitro culture medium induces.
2, according to the described construction process of claim 1, it is characterized in that: adding concentration when carrying out vitro culture in in-vitro culture medium is the external evoked mouse ovarian 48h-72h of 100-300nmol/L dexamethasone.
3, according to the described construction process of claim 2, it is characterized in that: adding concentration when carrying out vitro culture in in-vitro culture medium is the external evoked mouse ovarian 48h of dexamethasone of 300nmol/L.
4, according to the described construction process of claim 1, it is characterized in that: described in-vitro culture medium is the F-12 substratum, and its moiety comprises: 5%BSA, 0.1%DMSO, 10nmol/l Regular Insulin.
5, according to the described construction process of claim 1, it is characterized in that: described CO
2CO in the incubator
2Content be 5%, relative air humidity is 95%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2009101622985A CN101619304B (en) | 2009-08-03 | 2009-08-03 | Constructing method of ovary insulin resistant model |
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| CN2009101622985A CN101619304B (en) | 2009-08-03 | 2009-08-03 | Constructing method of ovary insulin resistant model |
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| CN101619304A true CN101619304A (en) | 2010-01-06 |
| CN101619304B CN101619304B (en) | 2011-05-18 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068439A (en) * | 2011-01-14 | 2011-05-25 | 吴效科 | Use of dexamethasone as tool medicament for constructing animal model with kidney-deficiency infertility |
| CN104705258A (en) * | 2015-02-10 | 2015-06-17 | 陕西师范大学 | Construction method and application of diet-induced insulin resistance model |
| CN108795846A (en) * | 2018-07-05 | 2018-11-13 | 江西科技师范大学 | For evaluating the cell model and evaluation method that improve insulin resistance product efficacy |
-
2009
- 2009-08-03 CN CN2009101622985A patent/CN101619304B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068439A (en) * | 2011-01-14 | 2011-05-25 | 吴效科 | Use of dexamethasone as tool medicament for constructing animal model with kidney-deficiency infertility |
| CN104705258A (en) * | 2015-02-10 | 2015-06-17 | 陕西师范大学 | Construction method and application of diet-induced insulin resistance model |
| CN104705258B (en) * | 2015-02-10 | 2018-01-05 | 陕西师范大学 | A kind of construction method of diet induced insulin resistant model and application |
| CN108795846A (en) * | 2018-07-05 | 2018-11-13 | 江西科技师范大学 | For evaluating the cell model and evaluation method that improve insulin resistance product efficacy |
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| CN101619304B (en) | 2011-05-18 |
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Inventor after: Wu Xiaoke Inventor after: Li Wei Inventor before: Wu Xiaoke |
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