The use of aspirin in the preparation of drugs for improving placenta previa or placental dysplasia.

Abstract

This invention provides the application of aspirin in the preparation of a drug for improving placenta accreta or placental dysplasia. This invention establishes an animal model of placenta accreta induced by uterine trauma and intervenes with high, medium, and low doses of aspirin after surgery. It was found that aspirin can improve excessive angiogenesis, increased perivascular inflammatory cell infiltration, excessive trophoblast invasion, and excessive formation of labyrinthine sinuses in mice induced by uterine trauma, and improve the immune regulation disorders in the uterus, decidua, and placental tissues caused by uterine trauma in mice. Based on the above research results, this invention applies aspirin to the preparation of a drug for improving placenta accreta or placental dysplasia, and the resulting drug can improve placenta accreta and placental dysplasia induced by uterine trauma.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the use of aspirin in the preparation of drugs for improving placenta previa or placental dysplasia. Background Technology

[0002] Placenta accreta is a pregnancy complication that occurs when placental villi abnormally invade the myometrium or even penetrate the serosa during delivery, preventing normal placental detachment and seriously endangering the mother's life. Clinical studies have found that uterine trauma is the most significant risk factor for placenta accreta. In the traumatic area, decidual defects allow the placental trophoblast and villi to penetrate the endometrium and invade the myometrium, and in severe cases, even reach surrounding pelvic organs. Simultaneously, trauma leads to local inflammation and abnormal angiogenesis, further promoting the abnormal invasion of placental villi. In recent years, due to the increasing global cesarean section rate, the incidence of placenta accreta has also risen accordingly, and the incidence increases significantly with the number of cesarean sections. Therefore, early diagnosis and timely intervention of placenta accreta are crucial for improving maternal and perinatal outcomes.

[0003] In recent years, with advancements in surgical techniques, the use of conservative surgical methods for placenta accreta, such as local lesion resection combined with uterine reconstruction, has increased significantly. Furthermore, research on targeted drug intervention is underway, including the use of methotrexate combined with mifepristone for placenta accreta patients and the adjunctive use of traditional Chinese medicine. However, methotrexate is a drug with serious toxic side effects and is generally used for postpartum placenta accreta in women who do not wish to have children, while traditional Chinese medicine has limited effectiveness in intervening in placenta accreta. Therefore, interventional drugs for placenta accreta or placental developmental abnormalities induced by uterine trauma are still under extensive exploration, and effective drug intervention techniques are currently lacking. Summary of the Invention

[0004] This invention provides the use of aspirin in the preparation of a drug for improving placenta accreta or placental dysplasia. By applying aspirin to the preparation of a drug for improving placenta accreta or placental dysplasia, the resulting drug can improve placenta accreta and placental dysplasia induced by uterine trauma.

[0005] According to a first aspect of the invention, the use of aspirin in the preparation of a medicament for improving placental implantation is provided.

[0006] One reason for the current lack of effective intervention methods for placenta accreta is the long-standing absence of mature animal models. Currently, placenta accreta animal models are mainly established using genetic engineering and surgical trauma methods. The surgical trauma method involves inducing placental developmental abnormalities in mice through perinatal cesarean section. In female mice after surgical trauma, increased trophoblastic cell invasiveness is observed in the placenta, the junctional layer boundary is unclear, and the number of invading trophoblastic cells in the decidua is significantly higher than in wild-type mice. Therefore, it can be used to simulate placenta accreta induced by cesarean section in clinical practice. Another reason is the strict restrictions on medication use for pregnant women; any drugs that may affect fertility or induce abnormal embryonic development are prohibited, thus greatly limiting the exploration of drug intervention techniques for this type of disease.

[0007] Aspirin belongs to the class of nonsteroidal anti-inflammatory drugs (NSAIDs). Its main pharmacological effects are antipyretic, analgesic, and anti-inflammatory, inhibiting the production of prostaglandins and thromboxanes through cyclooxygenases COX-1 and COX-2. Early inflammatory responses involve local vasodilation and increased capillary permeability, leading to the migration of leukocytes and macrophages to the inflammatory area; later, fibroblast proliferation occurs. NSAIDs can inhibit COX-2 at the site of inflammation, reducing prostaglandin synthesis and alleviating inflammation. Current research indicates that aspirin is relatively safe for pregnant women and does not increase the risk of congenital heart defects or developmental abnormalities in the embryo. Therefore, the current mechanism for using aspirin to prevent preeclampsia may involve improving the placental formation process and inhibiting platelet aggregation to stabilize endothelial cells, while simultaneously exerting antithrombotic and anti-inflammatory effects. However, whether aspirin can prevent placenta accreta or placental dysplasia has not yet been reported in studies.

[0008] The inventors of this application established a mouse model of placenta accreta by simulating a high-risk factor for placenta accreta—cesarean section—through surgical trauma. After surgery, the mice were treated with high, medium, and low doses of aspirin. Placental development was analyzed at 14.5 days of gestation. The results showed that feeding mice oral aspirin solution at concentrations of 0.5 g / L (medium dose) to 5 g / L (high dose) from uterine surgery until 14.5 days of gestation could improve the excessive maternal angiogenesis, increased perivascular inflammatory cell infiltration, excessive trophoblast invasion, and excessive formation of labyrinthine sinuses in the mouse placenta caused by uterine trauma. It also improved the immune regulation disorders in the uterine, decidual, and placental tissues caused by uterine trauma, thereby inhibiting placenta accreta induced by uterine trauma.

[0009] Based on the above research results, it is evident that aspirin can significantly inhibit placenta accreta induced by uterine trauma. Therefore, this application utilizes aspirin in the preparation of a drug to improve placenta accreta, and the resulting drug can significantly inhibit placenta accreta induced by uterine trauma. Since the use of aspirin to intervene in preeclampsia in pregnant women is a well-established practice, it is hoped that aspirin can be used in the preparation of a drug to improve placenta accreta and applied clinically to improve placenta accreta induced by uterine trauma. This has significant clinical implications for the intervention of placenta accreta.

[0010] Preferably, the placenta implantation is caused by uterine trauma.

[0011] Preferably, aspirin improves placenta accreta through at least one of the following pathways a to d: a. Inhibits excessive angiogenesis in the decidual layer; b. Reduce perivascular inflammatory cell infiltration in the decidual layer; c. Reduce excessive invasion of placental trophoblast cells; d. Reduce proliferation of blood sinuses in the labyrinth.

[0012] Preferably, the above-mentioned drug includes liquid and solid dosage forms, wherein the content of aspirin in the liquid dosage form is 0.5-5 g / L, and the content of aspirin in the solid dosage form is 0.1-1 mg / g.

[0013] Preferably, the above-mentioned liquid preparation is oral aspirin solution.

[0014] Preferably, the above-mentioned aspirin oral solution is prepared by the following steps: dissolving 0.5~5 g of aspirin powder in 1 L of autoclaved water to obtain the aspirin oral solution.

[0015] Studies have shown that oral administration of 0.5–5 g / L aspirin solution can inhibit placental implantation induced by uterine trauma in a mouse model.

[0016] According to a second aspect of the invention, a medicament for improving placenta accreta is provided, the medicament comprising aspirin; the medicament is a liquid or solid formulation, wherein the aspirin content in the liquid formulation is 0.5-5 g / L, and the aspirin content in the solid formulation is 0.1-1 mg / g.

[0017] This application applies aspirin to the preparation of a drug for improving placenta accreta. The prepared drug can significantly inhibit placenta accreta induced by uterine trauma, thereby achieving the purpose of improving placenta accreta induced by uterine trauma.

[0018] According to a third aspect of the invention, the use of aspirin in the preparation of a medicament for improving placental dysplasia is provided.

[0019] The inventors of this application discovered through experimental research that oral administration of aspirin to mice with a uterine trauma-induced placental implantation model can effectively reduce placental developmental abnormalities induced by uterine trauma in the mouse model.

[0020] Based on the above research results, it is evident that aspirin can significantly improve placental dysplasia induced by uterine trauma. Therefore, this application utilizes aspirin in the preparation of a drug for improving placental dysplasia. The resulting drug can effectively improve placental dysplasia induced by uterine trauma, and its clinical application can improve placental dysplasia induced by uterine trauma, which is of great significance for the clinical intervention of placental dysplasia.

[0021] Preferably, the aforementioned placental dysplasia is caused by uterine trauma.

[0022] Preferably, aspirin improves placental dysplasia through at least one of the following pathways a to d: a. Inhibits angiogenesis in the decidual layer; b. Reduce perivascular inflammatory cell infiltration in the decidual layer; c. Reduce the layer area of ​​the connecting layer; d. Reduce the layer area of ​​the labyrinth. Attached Figure Description

[0023] Figure 1 The figure shows the effects of uterine trauma and aspirin on pregnancy outcomes in late pregnancy in mice, as provided in Example 1.

[0024] Figure 2 The figure shows the effects of uterine trauma and aspirin on placental development in mice during late pregnancy, as provided in Example 2.

[0025] Figure 3 The figure shows the effects of uterine trauma and aspirin on placental trophoblast invasion and labyrinthine sinus formation in late pregnancy in mice, as provided in Example 3. Detailed Implementation

[0026] The technical features of the technical solution provided by the present invention will be further clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] Example 1: Effects of uterine trauma and aspirin on pregnancy outcomes in late pregnancy in mice This embodiment aims to establish an animal model of placenta accreta induced by uterine trauma through surgical trauma simulation, and to study the effects of uterine trauma and aspirin on pregnancy outcomes by administering medication from the postoperative period until 14.5 days of gestation. The specific experimental methods and procedures are as follows: 1. Using 7-8 week old SPF grade C57BL / 6J mice (approximately 18-20 g), a uterine trauma-induced placenta implantation animal model was established by simulating human uterine trauma through uterine surgery. That is, the mouse uterus was surgically induced to cause uterine trauma, thereby inducing placenta implantation. 2. Dissolve 5 g of aspirin (A5376-100G, Sigma-Aldrich) in dimethyl sulfoxide to prepare a stock solution with a concentration of 1 g / mL and store it at 4°C. When using, add drinking water and mix quickly. Dilute the prepared stock solution to prepare oral aspirin solutions with concentrations of 0.05 g / L (low dose), 0.5 g / L (medium dose), and 5 g / L (high dose). 3. Mice were given aspirin after surgery and continued to receive the medication after mating until late pregnancy, at which point the mice were sacrificed for anatomical observation. The experiment consisted of six groups: a normal control group (no surgery or aspirin treatment) and a sham surgery group (abdominal incision and suturing after anesthesia, without actual uterine surgery) with 8 mice in each group; and placenta accreta model mice were randomly divided into a simple surgery group (no aspirin treatment after surgery), a surgery plus low-dose group (administered 0.05 g / L aspirin oral solution after surgery), a surgery plus medium-dose group (administered 0.5 g / L aspirin oral solution after surgery), and a surgery plus high-dose group (administered 5 g / L aspirin oral solution after surgery), with 12 mice in each group. All mice had free access to water, medication, and food; the drinking water for the surgery and medication groups was the aspirin oral solution, which was changed daily, with each mouse drinking approximately 4 ml per day.

[0028] 4. At 14.5 days of gestation in mice, the pregnancy status in the uterus of each group of mice and the isolated embryos and placentas were observed. The morphology of the placenta was observed, and the embryo resorption rate, the proportion of embryos with delayed development, and the weight of the mouse fetuses were recorded. The results are as follows: Figure 1 As shown, where, Figure 1 The graph in Figure 'a' shows the pregnancy outcomes for the normal control group (CON), sham surgery group (Sham), surgery alone group (SUR), surgery plus low-dose aspirin group (Low Dose), surgery plus medium-dose aspirin group (Medium Dose), and surgery plus high-dose aspirin group (High Dose) (black arrows indicate embryo resorption). Figure 1 Figure b shows the effect of surgery on placental morphology in each group of mice. Figure 1The middle graph (c) shows a comparison of adverse pregnancy outcomes among the mouse groups. The left graph compares the embryo resorption rate among the mouse groups, the middle graph compares the proportion of delayed embryonic development among the mouse groups, and the right graph shows the weight distribution frequency of mouse fetuses (E14.5) isolated at 14.5 days of gestation. The dashed line represents the fifth percentile of fetal weight (0.1541g), N=12, *P<0.05, **P<0.01, ****P<0.0001 vs. normal control group.

[0029] Depend on Figure 1 It is known that pregnancy after uterine surgery in mice may lead to intrauterine bleeding and embryo resorption. Figure 1 Further statistical analysis revealed that compared with the normal control group (CON) and the sham surgery group (Sham), the embryo resorption rate and the proportion of embryonic developmental delay were increased in the simple surgery group (SUR) (embryo resorption rate: P<0.05). This indicates that surgical trauma may have an adverse effect on embryonic growth and development. After aspirin administration, the embryo resorption rate and the proportion of embryonic developmental delay decreased in the surgery plus medium-dose aspirin group (Medium Dose), while the surgery plus high-dose aspirin group (High Dose) showed even higher embryo resorption rate and the proportion of embryonic developmental delay (embryo resorption rate: P<0.01; proportion of embryonic developmental delay: P<0.0001). Figure 1 (c) Although the embryo resorption rate and the proportion of fetal growth retardation were higher in the surgery plus high-dose aspirin group than in the surgery alone group (SUR), the surgery plus high-dose aspirin group showed a stronger inhibitory effect on excessive trophoblast invasion and labyrinthine sinusoidal proliferation compared to the surgery plus medium-dose aspirin group. Therefore, different concentrations of oral aspirin solution differ in the specific changes (e.g., inhibition of excessive trophoblast invasion and labyrinthine sinusoidal proliferation, or improvement of embryo resorption rate and the proportion of fetal growth retardation).

[0030] The above results suggest that aspirin may have the effect of improving adverse pregnancy outcomes within a certain dosage range.

[0031] Example 2: Effects of uterine trauma and aspirin on placental development in late pregnancy in mice. This embodiment aims to establish an animal model of placenta accreta induced by uterine trauma through surgical trauma simulation, and to study the effects of uterine trauma and aspirin on placental development at 14.5 days of gestation (E14.5) by starting medication after surgery and continuing until late pregnancy. The specific experimental methods and procedures are as described in Example 1.

[0032] To analyze the effects of surgery and different concentrations of aspirin on the development of late-pregnancy placentas isolated from mice at 14.5 days of gestation, the E14.5 placenta was stained using hematoxylin-eosin (HE) staining. Furthermore, the connective and labyrinthic layers are important sites for material exchange between the mother and embryo, and changes in these layers may affect embryonic development. Therefore, this example also measured the area of ​​the connective and labyrinthic layers in the E14.5 placenta. The results are as follows: Figure 2 As shown, where, Figure 2 Image a shows the HE staining results of E14.5 placentas from mice in the normal control group (CON), sham-operated group (Sham), surgery-only group (SUR), surgery plus low-dose aspirin group (Low Dose), surgery plus medium-dose aspirin group (Medium Dose), and surgery plus high-dose aspirin group (High Dose). (The scale bar in the image is 100 μm long.) Red arrows indicate sites of angiogenesis, and the area within the dashed line represents the labyrinthine layer area. Figure 2 Figure b shows the measurement results of the layer area of ​​the connecting layer. Figure 2 In the middle, c represents the layer area measurement results of the labyrinth. Figure 2 In the middle d, the measurement results of the total placental area (the area of ​​the connecting layer + the area of ​​the labyrinth) are shown. N=12, #P<0.05, ##P<0.01, ###P<0.001 vs. normal control group.

[0033] Depend on Figure 2 HE staining results of E14.5 placentas from mice in the normal control group (CON), the surgery-only group (SUR), and the surgery-plus-low-dose aspirin group (LowDose) showed that the E14.5 placentas of the normal control group (CON) mice were clearly divided into three layers: the decidua basalis (DB, the region between the myometrium and the junctional layer, adjacent to the myometrium and junctional layer, including the area indicated by the red arrow), the junctional zone (JZ), and the labyrinthal zone (LA), with regularly distributed cells around blood vessels. In contrast, the placentas of the surgery-only group (SUR) and the surgery-plus-low-dose aspirin group (LowDose) showed significantly different distributions. In E14.5 mice with surgical intervention (Dose), angiogenesis was observed in the decidual layer of the placenta, along with extensive inflammatory cell infiltration around the blood vessels (the "white bubbles" represent blood vessels in the decidual layer, surrounded by a ring of darkly stained cells representing inflammatory cells). These results suggest that surgery may induce an inflammatory response within the placenta, promoting excessive maternal angiogenesis. HE staining of E14.5 placentas from mice in the Medium Dose and High Dose groups showed that aspirin administration significantly improved angiogenesis and inflammatory cell infiltration. Figure 2(a) The area measurements of the connective and labyrinthic layers showed that, compared with the normal control group (CON), the labyrinthic layer area in the E14.5 placenta of mice in the simple surgery group (SUR) was increased, and the difference in the area of ​​the connective and labyrinthic layers between the simple surgery group (SUR) and the normal control group (CON) did not reach statistical significance. However, aspirin administration significantly reduced the area of ​​the connective layer (low-dose group: P<0.05, high-dose group: P<0.01) and the labyrinthic layer (low-dose group and high-dose group: P<0.01), with the most significant effect observed in the surgery plus high-dose aspirin group (High Dose). Figure 2 (b, c, d) and, as can be seen from the above, moderate doses of aspirin can inhibit the increase in the area of ​​the placental junction layer and labyrinthine layer after surgery.

[0034] Placental dysplasia refers to abnormalities in the morphology and size of the placenta. Compared to the normal control group (CON), increased placental area, increased area of ​​the connective and labyrinthal layers, and increased vascularity may lead to placental overgrowth and placenta accreta. These results demonstrate that aspirin can mitigate the adverse effects of uterine trauma on placental development.

[0035] Example 3: Effects of uterine trauma and aspirin on trophoblast invasion and LA sinusoid formation in the placenta during late pregnancy Cytokeratins are a family of proteins that make up the cytoskeleton (intermediate filaments) and are found in epithelial cells. Cytokeratin 7 (CK7) is a specific member of this family. CK7 is a marker of various glandular epithelium and transitional epithelium. Trophoblasts (cytotrophoblasts and syncytiotrophoblasts) on the surface of placental villi strongly express CK7.

[0036] CD34 is a glycoprotein found on the surface of cell membranes. It is a very commonly used and sensitive marker for vascular endothelial cells, and all vascular endothelial cells (a special type of epithelial cell) express CD34.

[0037] Based on the HE staining results of Example 2, this example performed immunohistochemical staining with trophoblast marker CK7 and vascular epithelial cell marker CD34 to assess the invasion of trophoblasts and the formation of labyrinthine sinusoids in the placenta at 14.5 days of gestation (E14.5). The results are as follows: Figure 3 As shown, where, Figure 3Figure 'a' shows the immunohistochemical staining results of CK7 in trophoblast cells from the E14.5 placenta of mice in the normal control group (CON), sham-operated group (Sham), surgery-only group (SUR), surgery plus low-dose aspirin group (Low Dose), surgery plus medium-dose aspirin group (Medium Dose), and surgery plus high-dose aspirin group (High Dose). The area within the dashed line represents the trophoblast cell invasion area. The extent to which CK7-positive cells invade outward into the maternal myometrium is defined as the trophoblast cell invasion area. Figure 3 Image b shows the CD34 immunohistochemical staining results of labyrinthine sinusoids in the E14.5 placenta of each group of mice. The red arrows indicate the sites of labyrinthine sinusoid formation. Figure 3 Figure c shows the statistical results of trophoblast invasion area in the E14.5 placenta of each group of mice. Figure 3 The figure in the middle d is a statistical result of the number of labyrinthine blood sinuses generated in the E14.5 placenta of each group of mice (number of sinuses). N=12, #P<0.05, ####P<0.0001 vs. SUR group.

[0038] Depend on Figure 3 It was found that the trophoblast invasion area in the E14.5 placenta of the surgically operated (SUR) group mice was increased, while different concentrations of aspirin could reduce the degree of trophoblast invasion, showing a dose-response effect. Figure 3 In the medium-dose group (a), c, and high-dose group: P<0.0001); similarly, the statistical results of CD34-positive sinusoids showed that postoperative pregnancy increased the formation of labyrinthine sinusoids, while aspirin could inhibit the formation of labyrinthine sinusoids ( Figure 3 (Medium-b, d, and high-dose groups: P<0.05).

[0039] The above results demonstrate that aspirin improves excessive trophoblast invasion and excessive formation of labyrinthine sinusoids after uterine trauma.

[0040] In summary, aspirin administration reduced perivascular inflammatory cell infiltration, trophoblast invasion, and angiogenesis in the decidual layer of the placenta, as well as decreased sinusoidal hyperplasia in the labyrinth, exhibiting a dose-response effect. Therefore, oral aspirin can significantly inhibit placenta implantation induced by uterine trauma and improve placental dysplasia induced by uterine trauma.

[0041] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention, but such modifications or substitutions are all within the scope of protection of the present invention.

Claims

1. The use of aspirin in the preparation of drugs for improving placental implantation.

2. The use of aspirin as described in claim 1 in the preparation of a medicament for improving placenta accreta, characterized in that: The placenta accreta was caused by uterine trauma.

3. The use of aspirin as described in claim 1 in the preparation of a medicament for improving placenta accreta, characterized in that, The aspirin improves placenta accreta through at least one of the following pathways a to d: a. Inhibits excessive angiogenesis in the decidual layer; b. Reduce perivascular inflammatory cell infiltration in the decidual layer; c. Reduce excessive invasion of placental trophoblast cells; d. Reduce proliferation of blood sinuses in the labyrinth.

4. The use of aspirin as described in claim 1 in the preparation of a medicament for improving placenta accreta, characterized in that: The drug includes liquid and solid dosage forms, wherein the aspirin content in the liquid dosage form is 0.5-5 g / L, and the aspirin content in the solid dosage form is 0.1-1 mg / g.

5. The use of aspirin as described in claim 4 in the preparation of a medicament for improving placenta accreta, characterized in that: The liquid preparation is oral aspirin solution.

6. The use of aspirin as described in claim 5 in the preparation of a medicament for improving placenta accreta, characterized in that, The aspirin oral solution is prepared by the following steps: dissolving 0.5-5 g of aspirin powder in 1 L of autoclaved water to obtain the aspirin oral solution.

7. A drug for improving placenta accreta, characterized in that: The medications include aspirin; The drug is a liquid or solid dosage form, wherein the aspirin content in the liquid dosage form is 0.5-5 g / L, and the aspirin content in the solid dosage form is 0.1-1 mg / g.

8. The use of aspirin in the preparation of drugs for improving placental dysplasia.

9. The use of aspirin as described in claim 8 in the preparation of a medicament for improving placental dysplasia, characterized in that: The placental malformation was caused by uterine trauma.

10. The use of aspirin as described in claim 8 in the preparation of a medicament for improving placental dysplasia, characterized in that, The aspirin improves placental dysplasia through at least one of the following pathways a to d: a. Inhibits angiogenesis in the decidual layer; b. Reduce perivascular inflammatory cell infiltration in the decidual layer; c. Reduce the layer area of ​​the connecting layer; d. Reduce the layer area of ​​the labyrinth.

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