CN114451358B - Animal model for preeclampsia diseases and construction method and application thereof - Google Patents
Animal model for preeclampsia diseases and construction method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
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- A01K67/0276—Knock-out vertebrates
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01K2267/03—Animal model, e.g. for test or diseases
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- A01K2267/03—Animal model, e.g. for test or diseases
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Abstract
The invention discloses an animal model for preeclampsia diseases, which uses Sirt1 +/‑ The pregnant mouse heterozygote is knocked out systemically and is an animal model of preeclampsia diseases. The invention also discloses a construction method and application thereof. The preeclampsia disease animal model does not need external medicine and operation intervention, does not need special feed and special feeding environment for reproduction, can completely reduce preeclampsia diseases, can be used for helping to clarify the pathogenesis and mechanism of the preeclampsia diseases and can also be used for helping to screen and evaluate the efficacy of the medicines for preventing and treating the preeclampsia diseases.
Description
Technical Field
The invention relates to the technical field of animal model construction, in particular to an animal model for preeclampsia diseases and a construction method and application thereof.
Background
Preeclampsia (PE) is a specific multisystemic progressive disease during pregnancy, mainly manifested as late gestational or postpartum onset of new hypertension and proteinuria; or new onset hypertension and significant end organ dysfunction, with or without urinary protein. The method is characterized in that the method often causes poor pregnancy outcome of the mother and the child and increases the occurrence risk of long-term cardiovascular diseases, the current cause is unknown, and no effective treatment means exists. However, since clinical symptoms subside after delivery, recent guidelines are attributed to placental and maternal vascular dysfunction.
An ideal animal model should present the clinical features of the disease. PE, which is specific for the middle and late gestation, manifests as insufficient trophoblast infiltration, recanalization of spiral arteries, concomitant hypertension, diffuse vascular endothelial dysfunction, and further causes kidney damage, urinary protein, and other end organ dysfunction. Because the pathogenic factors are unknown and the overall disease symptoms are wide, the currently known models cannot completely present all symptoms of PE. At present, common rat and mouse models comprise an inflammation model induced by lipopolysaccharide, a vascular contraction model induced by RUPP (vascular angioplasty-placental perfusion pressure) and an anti-angiogenesis model induced by soluble fms-like tyrosine kinase 1 and the like, and the common rat and mouse models focus on blood pressure and urine protein signs, cannot well reduce the course of diseases, cannot give consideration to trophoblast invasion disorder and terminal organ dysfunction such as kidney specific to preeclampsia, and cannot reduce the clinical characteristics of the diseases. And the molding cost is higher, the operation is more complicated, and the individual difference of experiment operators exists no matter the medicine dosage or the operation mode, thereby influencing the molding success rate.
Sirt1 is a Nicotinamide Adenine Dinucleotide (NAD) + ) The dependent protein deacetylase is a homolog of Sir2 protein in mammals, and belongs to the family of mammalian Sirtuins proteins. Current studies suggest that Sirt1 is distributed in both the nucleus and cytoplasm. Sirt1 exerts anti-inflammatory, antioxidant stress, anti-aging, T cell immunity activation and even mitochondrial generation by deacetylating lysine residues of various proteins.
Aiming at a series of technical problems of high molding cost, complicated molding process and pathophysiology process which can not be completely recovered of model animals, the invention provides a novel animal model, sirt1 +/- The heterozygote state is knocked out to restore the pathophysiological process of the disease, and the pregnancy state is relieved or the treatment with agonist can be recovered. In addition, external medicines and surgical intervention are not needed, and special feed and special feeding environment are not needed for propagation, so that a brand-new animal model is provided for the research of the preeclampsia morbidity process and mechanism.
Disclosure of Invention
The invention aims to provide an animal model for preeclampsia diseases and a construction method and application thereof, so as to solve the defects of the prior art.
The invention adopts the following technical scheme:
the first aspect of the invention provides the use of the Sirt1 gene in the construction of an animal model of preeclampsia disease.
In a second aspect, the invention provides an animal model of preeclampsia diseases, using Sirt1 +/- The pregnant mouse with heterozygote knocked out systemically is an animal model with preeclampsia diseases.
Further, the Sirt1 +/- The whole-body knockout heterozygote pregnant mouse is obtained by the following steps: mixing Sirt1 flox/flox The gene editing mice were propagated and crossed with Dppa3-IRES-Cre mice and bred via decrepit.
The third aspect of the invention provides a method for constructing an animal model of preeclampsia diseases, which comprises the following steps: sirt1 is flox/flox After the gene editing mouse is propagated, the gene editing mouse is hybridized with a Dppa3-IRES-Cre mouse, and after Cre removal breeding, sirt1 is obtained +/- And (3) knocking out the heterozygous pregnant mouse in the whole body to obtain the constructed preeclampsia disease animal model.
The fourth aspect of the invention provides the application of the preeclampsia disease animal model in researching the pathogenesis and mechanism of preeclampsia diseases.
The fifth aspect of the invention provides application of the preeclampsia disease animal model in screening of drugs for preventing and treating preeclampsia diseases and evaluation of drug effects.
The sixth aspect of the invention provides application of the Sirt1 gene in preparing a medicament for preventing and treating preeclampsia diseases.
The invention has the beneficial effects that:
the invention provides a novel preeclampsia disease animal model which does not need external medicine and operation intervention, does not need special feed and special feeding environment for propagation, and can completely reduce preeclampsia diseases.
According to the method, the Sirt1 gene expression of the target animal is partially knocked out, so that the target animal shows more typical preeclampsia characteristics, the rise of blood pressure in the late gestation period is spontaneously formed, the target animal recovers after delivery, and fetal rat genes and the Sirt1 gene are found +/- The pregnant mouse has relativity to the blood pressure difference value in the early and late gestation period; in addition, sirt1 is found in the late gestation +/- The proportion of the placenta functional layer of pregnant mouse is reduced, indicating that the placenta is nourishingDysfunction such as cell invasion and spiral artery recasting, and proteinuria and nephropathy appear; under the action of the gene agonist, relevant pathological manifestations can be restored, the successful construction of a disease model is prompted, and the fact that the gene is one of the pathogenic genes in preeclampsia is demonstrated.
The preeclampsia disease animal model can be used for helping to clarify the pathogenesis and mechanism of preeclampsia diseases and can also be used for helping to screen and evaluate the drug effect of the preeclampsia diseases.
Drawings
FIG. 1A is Sirt1 +/- Schematic diagram of construction of a whole-body knockout heterozygote pregnant mouse.
FIG. 1B shows PCR agarose gel electrophoresis analysis of Sirt1 +/- The genotype of the offspring of the heterozygous pregnant mouse (fetal mouse, E18.5: 18.5 days of pregnancy) is knocked out in a whole body. In the figure, M is a DNA indicator band, WT is Sirt1 flox/flox Pregnant mouse tail control, KO knock-out gene band indicated size: 500bp, HO: homozygote, HE: heterozygote, WT wild gene band indicates size: 900bp. B33-1, B33-2 and B33-4 are heterozygote Sirt1 +/- B33-3 and B33-5 are homozygote Sirt1 -/- 。
FIG. 1C is Sirt1 +/- Group sum Sirt1 flox/flox Immunofluorescence of Sirt1 in group placenta (E18.5) tissue. CK7 marks trophoblasts and DAPI marks the nucleus. Sirt1 can be seen +/- The expression of the trophoblast Sirt1 in the placenta is obviously reduced, and the knockout efficiency is verified.
FIG. 1D is Sirt1 +/- Group sum Sirt1 flox/flox Appearance of mice (E18.5) and placenta (E18.5). Sirt1 can be seen +/- Both fetal rat and placenta of group were less than Sirt1 flox/flox And (4) grouping.
FIG. 1E is Sirt1 +/- Group sum Sirt1 flox/flox Group embryo uptake rate. Sirt1 +/- Groups had an ascending trend but no statistical difference.
FIG. 1F is Sirt1 +/- Group sum Sirt1 flox/flox Group placenta (E18.5) weight. Sirt1 +/- Groups showed no statistical difference, although they tended to decline.
FIG. 1G shows Sirt1 +/- Group sum Sirt1 flox/flox Group fetal rat (E18.5) weight. Sirt1 +/- Group comparison Sirt1 flox/flox The groups decreased significantly.
FIG. 1H shows Sirt1 +/- Group sum Sirt1 flox/flox The gene ratios of the mice in groups of E18.5 and P28. E18.5: day 18.5 of gestation, P28: postnatal 28 days, HO: homozygote, HE: heterozygote, WT: and (4) a wild type. After 28 days of birth, the survival rate of the homozygote is extremely low, and the lethal phenomenon of the homozygote in the perinatal period is verified.
FIG. 1I is Sirt1 +/- Group sum Sirt1 flox/flox Blood pressure of each pregnancy period of the pregnant mice. Basic condition: basal status (non-pregnant period), earlyPG: early pregnancy, midPG: middle trimester, latex g: in the late gestation period. Sirt1 can be seen +/- Blood pressure of pregnant mice in the late pregnancy is obviously increased.
FIG. 1J shows Sirt1 +/- Group sum Sirt1 flox/flox Blood pressure difference of early and late gestation of the pregnant rats. Sirt1 can be seen +/- The blood pressure in the late pregnancy of the group shows a remarkably rising trend compared with the blood pressure in the early pregnancy.
FIG. 1K is Sirt1 +/- And (3) analyzing the correlation between the blood pressure difference of the pregnant mouse in the early and late gestation period and the ratio of a knockout gene of a fetal mouse (E18.5), namely the Sirt1 gene. Therefore, the proportion of the knockout gene in fetal mice and the blood pressure difference of pregnant mice in the early and late pregnancy have certain linear relation, and the statistical significance is achieved.
FIG. 1L is Sirt1 +/- Group sum Sirt1 flox/flox Masson staining and PAS staining of kidney tissue of gestating mice (E18.5). Sirt1 can be seen +/- The endothelial cells of glomerulus capillaries in the later gestation period of the pregnant mice are subjected to diffuse hyperplasia and swelling, and the capillary lumen is narrow, so that the kidney lesion in the preeclampsia is more typical.
FIG. 1M is Sirt1 +/- Group sum Sirt1 flox/flox Urine protein concentration of pregnant mice (E17.5-E18.5). Sirt1 can be seen +/- The urine protein of pregnant mice in the late gestation period is obviously increased.
FIG. 1N is Sirt1 +/- Group sum Sirt1 flox/flox Placental longitudinal sections of pregnancies rats (E18.5). Labyrinth layer: is composed of maternal blood sinus and villus, mediates the exchange of nutrition, gas and metabolic waste between the maternal and the fetus, and is the function of the placenta of pregnant miceEnergy layer, connecting region: i.e., the sponge layer, has primarily endocrine function, often indicating the placenta active functional area as the labyrinth/connective area.
Fig. 1O is a statistical plot of the placental labyrinth layer/junction area described above. Sirt1 can be seen +/- The active functional area of the pregnant mouse placenta is more than Sirt1 flox/flox Pregnant mice were significantly narrower.
FIG. 1P is Sirt1 +/- Group sum Sirt1 flox/flox Blood pressure of pregnant mice in late gestation period and postpartum blood pressure. E17/18: blood pressure at day 17.5 of gestation, P2/3: 2.5 days after delivery. Sirt1 can be seen +/- Pregnant mice recover from elevated blood pressure after delivery.
FIG. 1Q is Sirt1 +/- Group sum Sirt1 flox/flox The postpartum urine protein (P2.5-P3.5) of the pregnant mice. Sirt1 can be seen +/- Pregnant mice recover from elevated urine protein after delivery.
Figure 2A is a dosing regimen. After embolus appears in 0.5 day of pregnancy, sirt1 is added +/- Pregnant mice were randomly divided into two groups, SRT2104 group and Vehicle group, from E1.5 to E17.5 (day 1.5 to day 17.5 of pregnancy), SRT2104 group was intraperitoneally injected with SRT2104 200mg/kg every other day, and Vehicle group was injected with an equal amount of solvent.
Fig. 2B shows the appearance of fetal rats (E18.5) and placenta (E18.5) in the Vehicle group and SRT2104 group. The SRT2104 group of fetal mice significantly increased over the Vehicle group.
FIG. 2C shows the embryo uptake rates for the Vehicle group and the SRT2104 group. There was a trend of decreasing uptake of SRT2104 group embryos, but there was no statistical difference.
FIG. 2D shows placenta (E18.5) weights of the Vehicle group and the SRT2104 group. The placenta of SRT2104 group was heavier than the Vehicle group.
FIG. 2E shows the weights of fetal mice (E18.5) from the Vehicle group and the SRT2104 group. The SRT2104 group of fetal mice was heavier than the Vehicle group.
FIG. 2F shows the blood pressure of pregnant mice in both the Vehicle group and the SRT2104 group during pregnancy. Basic condition: basal status (non-pregnant period), earlyPG: early pregnancy, midPG: middle trimester, latex g: in the late gestation period. Showing Sirt1 after administration of SRT2104 +/- The blood pressure of pregnant mice in the late gestation period due to gene increase is corrected.
FIG. 2G shows pregnancy of early and late pregnant Vehicle group and SRT2104 groupBlood pressure difference. It can be seen that after administration of SRT2104, sirt1 +/- The blood pressure of pregnant mice in the late gestation period due to gene increase is corrected.
FIG. 2H shows the urinary protein levels of pregnant mice in the Vehicle group and SRT2104 group (E17.5-E18.5). Showing Sirt1 after administration of SRT2104 +/- The urinary protein of pregnant mice in the late gestation period due to gene increase is corrected.
FIG. 2I shows Masson staining and PAS staining of kidney tissue of pregnant mice (E18.5) in the Vehicle group and SRT2104 group. It can be seen that after administration of SRT2104, sirt1 +/- The kidney lesion caused by the gene in the late gestation period of the pregnant mouse is corrected.
FIG. 2J shows placental longitudinal sections of pregnant mice (E18.5) from both the Vehicle group and the SRT2104 group.
Fig. 2K is a statistical map of the placental labyrinth layer/junction area described above. Suggesting that SRT2104 post-dose Sirt1 +/- The labyrinthic layer of pregnant mice in the late gestation period caused by gene constriction is corrected.
Detailed Description
The invention is explained in more detail below with reference to exemplary embodiments and the accompanying drawings. The following examples are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
1 preeclampsia disease animal model: sirt1 +/- Construction of whole-body knockout heterozygote pregnant mouse
As required by subsequent experiments, the promoter-driven Cre-loxP system is adopted in the invention. Sirt1 was introduced from JAX (The Jackson Laboratory) flox/flox Gene editing mice (B6.129-Sirt 1) tm3Fwa /DsinJ). And order Dppa3-IRES-Cre mice from Nanmo Biometrics (Shanghai, square model Biotechnology, inc.), because Dppa3-Cre can effectively exert Cre recombinase activity in early embryo and germ cell lines, the mice are effective commercialized tool mice for constructing a whole-body knockout model. Commissioned to Nanmo Biometrics for crossing and propagation via De-Cre to yield Sirt1 +/- Heterozygous pregnant mice were knocked out systemically and the mating strategy is shown in figure 1A:
1) Introduction of Sirt1 from JAX flox/flox Gene editing mice (B6.129-Sirt 1) tm3Fwa DsinJ, numbering: 029603, also known as Sirt1 flox/flox Mouse).
2) Order commercially available Dppa3-IRES-Cre mice from south mous biology.
3) Let Sirt1 flox/flox Mouse and wild type Sirt1 +/+ Hybridizing and propagating the mice to obtain Sirt1 flox/+ A mouse.
4) Sirt1 is flox/+ The mice were crossed with Dppa3-IRES-Cre mice to obtain Sirt1 flox/+ DPPA3-CRE + A mouse.
5) Sirt1 is flox/+ DPPA3-CRE + Mouse and wild type Sirt1 +/+ Hybridizing the mice, removing DPPA3-CRE gene, further expanding propagation to obtain Sirt1 +/- Heterozygote mice were knockout systemically.
6) Sirt1 is +/- Knockout heterozygote mice to get Sirt1 +/- Whole-body knockout heterozygote pregnant mouse (hereinafter Sirt 1) +/- Pregnant mice) i.e. the preeclampsia disease animal model.
7).Sirt1 +/- Offspring can be obtained by breeding pregnant mice, and the genotype is represented as follows: 1/2 heterozygote: sirt1 +/- 1/4 wild type: sirt1 flox/flox And 1/4 homozygote: sirt1 -/- (less survival in perinatal period). Mixing Sirt1 flox/flox Mice are caged to obtain Sirt1 flox/flox And (5) pregnant mice.
8) Verifying fetal mouse genotype through PCR agarose gel electrophoresis, and verifying Sirt1 protein expression down-regulation in placenta tissues through immunofluorescence technology, as shown in figure 1B and figure 1C; and confirmed literature reports of homozygote fetal rat perinatal lethality, as shown in FIG. 1H.
2 the pre-eclampsia disease animal model constructed by the method has pre-eclampsia-like symptoms
The invention further carries out systematic evaluation on preeclampsia related performance of the constructed preeclampsia disease animal model, and comprises the steps of collecting 24h urine of a pregnant mouse, observing basic conditions of pregnancy blood pressure, fetal mouse size, fetal mouse weight, embryo absorption rate of the pregnant mouse and the like, carrying out cervical dislocation and sacrifice after 18.5 days of pregnancy, and taking kidney and placenta tissues for further morphological observation. Randomly selecting Sirt1 in the same batch grouping +/- The pregnant mouse is remained until parturition, and the blood pressure recovery and urine protein change are observedGood condition and the like. The heterozygote group, sirt1 +/- The pregnant mice of the group were Sirt1 +/- Pregnant mice, control group Sirt1 flox/flox The pregnant mice of the group were Sirt1 flox/flox And (5) pregnant mice.
We have found Sirt1 +/- Embryo uptake rates were almost normal in pregnant mice (fig. 1E). The weight difference between the two placenta groups was not statistically significant (FIG. 1F, sirt1 +/- Sirt1, group vs flox/flox Groups). Since preeclampsia can cause Fetal Growth Restriction (FGR), we analyzed Sirt1 flox/flox Group sum Sirt1 +/- The live carcasses of the group were heavy and Sirt1 was found +/- The live carcass weight of the group was significantly reduced (FIG. 1G, sirt1 +/- Sirt1, group vs flox/flox Group (2): 0.7803 ± 0.1651vs.0.8559 ± 0.1585 g). Representative images of groups of fetal mice and placenta are shown in FIG. 1D, along with Sirt1 flox/flox Group comparison, sirt1 +/- The group born mice were small. Subsequently, we analyzed the change in Systolic Blood Pressure (SBP). Interestingly, sirt1 +/- The level of SBP in the late gestation of the pregnant mice is obviously increased (figure 1I, sirt1) +/- Sirt1 group vs flox/flox Group (2): 119.6 +/-9.952vs.108 +/-6.340 mmHg). To understand the level of blood pressure rise, we calculated the difference Δ BP between the late-gestation systolic pressure and the early-gestation systolic pressure. We have found that Δ BP is at Sirt1 +/- Higher levels were present in the group (FIG. 1J, sirt1 +/- Sirt1, group vs flox/flox Group (2): 12.45 + -9.186 vs. -1.562 + -10.47 mmHg). Interestingly, Δ BP levels were positively correlated with Sirt1 gene values in fetal mice (fig. 1K). Sirt1 +/- After delivery, blood pressure and urine protein levels of pregnant mice were recovered, as shown in fig. 1P and 1Q.
Renal injury is another characteristic of preeclampsia. Typical manifestations are urine protein and renal pathologies, including glomerular enlargement and reduced blood supply, due to endothelial cell swelling, with associated membrane cells and capillary lumen occlusion. In this study, we examined Sirt1 +/- Concentration of urine protein in early and late gestation of pregnant mice, sirt1 +/- The concentration of urine protein in the late gestation period of pregnant mice is obviously increased (figure 1M, sirt1) +/- Sirt1, group vs flox/flox Group (2): 2.316 ± 0.05245vs.2.189 ± 0.05252 ug/ml). We also used MassonStaining and PAS staining the pathological changes of the kidneys of each group were observed. And Sirt1 flox/flox Group comparison, sirt1 +/- The gestating mice exhibited typical preeclampsia-associated glomerular injury (FIG. 1L). In addition, placental abnormalities are a substantial change in patients with preeclampsia. In our mouse model, we observed Sirt1 in Masson staining results +/- Labyrinth layer is compared with Sirt1 flox/flox Narrow (fig. 1N). We measured the labyrinth/junction ratio of each placenta group and found Sirt1 +/- Group and Sirt1 flox/flox The labyrinthine layer/junction area ratio decreased significantly compared to the group (fig. 1O). The labyrinth layer is a dense structure formed by trophoblasts and associated fetal blood vessels undergoing extensive villous branching. The labyrinthine layer is the functional layer of the placenta, and narrowing of the labyrinthine layer means dysfunction of the placenta. However, as we previously mentioned, there was no significant difference in the weight of the two groups of placentas (fig. 1F), which is likely due to the difference in handling caused by the residual decidua of part of the placentas at the time of primary exfoliation.
Sirt1 +/- Typical preeclampsia-like symptoms appear in the pregnancy of pregnant mice, and are shown as hypertension, urine protein, low fetal weight, kidney injury and reduction of proportion of placenta labyrinth layer, so that successful construction of a preeclampsia-like mouse model is prompted.
The 3 Sirt1 agonist SRT2104 significantly improved Sirt1 +/- Preeclampsia-like symptoms of pregnant mice
Furthermore, we have experimentally found that supplementation with Sirt1 agonists can reverse the onset of the above pathological manifestations. To verify the effect of Sirt1 in preeclampsia, we injected SRT2104 200mg/kg (working solution concentration 3 mg/ml) intraperitoneally every other day from E1.5 to E17.5 (day 1.5 to day 17.5 of pregnancy) to Sirt1 +/- Pregnant mice, as SRT2104 group. Vehicle (solvent) group injected with equal amount of solvent (solvent is solvent of SRT 2104: 5% DMSO +40% PEG300+5% The% Tween80+50% 2 O), as a control group. The course of treatment of mice is shown in figure 2A.
Administering SRT2104 treatment for Sirt1 +/- After pregnancy, there was no significant change in embryo uptake rate (FIG. 2C), a slight increase in placenta weight (FIG. 2D, vehicle group vs. SRT2104 group: 0.08519. + -. 0.01009vs. 0.09237. + -. 0.01213 g), a significant increase in fetal mouse weight (FIG. 2E,set Vehicle group vs. srt 2104: 0.6808 ± 0.08630vs.0.7719 ± 0.1483 g). These results are consistent with representative images of fetal mice and placenta (fig. 2B). It is clear that both the late gestational systolic blood pressure levels in the SRT2104 group (FIG. 2F, vehicle group vs. SRT2104 group: 117.8. + -. 8.311vs. 107.4. + -. 10.21 mmHg) and Δ BP (FIG. 2G, vehicle group vs. SRT2104 group: 8.158. + -. 9.212vs. -5.234. + -. 12.86 mmHg) are significantly reduced compared to the Vehicle group. SRT2104 ameliorated kidney injury, decreased urine protein concentration in late gestation (FIG. 2H, vehicle group vs. SRT2104 group: 7.397 + -3.293vs.3.392 + -0.7711 ug/ml), and glomerular morphology was essentially normal (FIG. 2I). Following intraperitoneal injection using SRT2104, the placenta also showed labyrinthine layer broadening (fig. 2J-2K).
These results demonstrate that increased Sirt1 expression can reverse preeclampsia-like behavior, further demonstrating that Sirt1 is an important causative factor in preeclampsia.
Claims (4)
1. An animal model of preeclampsia diseases is characterized in that Sirt1 is used +/- The pregnant mouse with heterozygote knocked out systemically is an animal model with preeclampsia diseases, and Sirt1 is +/- The whole-body knockout heterozygote pregnant mouse is obtained by the following steps: mixing Sirt1 flox/flox The gene editing mice were propagated and crossed with Dppa3-IRES-Cre mice and bred via decrepit.
2. The construction method of the animal model of the preeclampsia diseases is characterized by comprising the following steps: mixing Sirt1 flox /flox After propagation, the gene editing mice are hybridized with Dppa3-IRES-Cre mice, and after Cre removal propagation, sirt1 is obtained +/- And (3) knocking out the heterozygous pregnant mouse in the whole body to obtain the constructed preeclampsia disease animal model.
3. Use of the preeclampsia disease animal model of claim 1 in studying the pathogenesis and mechanism of preeclampsia disease.
4. The use of the preeclampsia disease animal model of claim 1 in screening drugs for preventing and treating preeclampsia diseases and evaluating drug effects.
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