Application of flavonoid compound fissistigma factor in preparation of intrauterine adhesion treatment drugs

Abstract

The application discloses application of flavonoid compound quercetin in preparation of a drug for treating intrauterine adhesion, and belongs to the technical field of biological medicines. It is found that the flavonoid compound quercetin can relieve the inflammation and fibrosis degree of the uterus of a mouse with intrauterine adhesion, which indicates that the quercetin can relieve the intrauterine adhesion disease process and has an application prospect of developing an anti-intrauterine adhesion treatment drug.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of the flavonoid compound phloem in the preparation of drugs for the treatment of intrauterine adhesions. Background Technology

[0002] Intrauterine adhesions (IUA), also known as Asherman's syndrome, are caused by damage to the endometrium, leading to partial or complete closure of the uterine cavity. Essentially, it is endometrial fibrosis. After the endometrium is damaged, pathogens that have infiltrated the wound site release damaging or pathogen-associated molecular patterns, and damaged cells release chemokines, triggering inflammation. If the inflammation persists, it will lead to endometrial fibrosis.

[0003] Intrauterine adhesions causing partial endometrial dysfunction can lead to recurrent pregnancy loss, oligomenorrhea accompanied by pelvic pain, amenorrhea, and infertility, seriously affecting human reproductive health. In recent years, with the increase in clinical intrauterine procedures, the incidence of intrauterine adhesions has also been rising, becoming the second leading cause of secondary infertility in women. Currently, commonly used treatments include hysteroscopic adhesiolysis, intrauterine device (IUD) placement, and hormone therapy; however, these methods have a high recurrence rate and cannot alleviate endometrial fibrosis.

[0004] Therefore, there is an urgent need to find new drugs to treat intrauterine adhesions and achieve endometrial reconstruction, so as to provide new ideas and methods for the clinical treatment of intrauterine adhesions. Summary of the Invention

[0005] The purpose of this invention is to provide the application of the flavonoid compound phloem in the preparation of drugs for the treatment of intrauterine adhesions.

[0006] The flavonoid compound, pectin, has the chemical name 5,7-dihydroxy-6-methoxy-2-phenyl-4H-1-benzofuran-4-one, and its structural formula is shown below:

[0007] .

[0008] In one embodiment of the present invention, the effects of low-dose (20 mg / kg) and high-dose (40 mg / kg) phloem on endometrial morphology and inflammation levels in mice with intrauterine adhesions were determined by HE staining, ELISA and qRT-PCR experiments.

[0009] In another embodiment of the present invention, the effects of low-dose (20 mg / kg) and high-dose (40 mg / kg) phloem on the degree of endometrial fibrosis in mice with intrauterine adhesions were determined by Masson staining and α-SMA immunohistochemistry.

[0010] This invention has found that the flavonoid compound styrax can improve the morphology of the endometrium in mice with intrauterine adhesions and alleviate its inflammation and fibrosis, indicating that styrax can alleviate the progression of intrauterine adhesions and has the potential to be used in the development of anti-intrauterine adhesions treatment drugs. Attached Figure Description

[0011] Figure 1 The results of HE staining of uterine tissue (A), expression of inflammatory factor genes (B), and expression of inflammatory factors in serum (C) are shown in Example 1 under different concentrations of phloem.

[0012] Figure 2 The results of Masson staining of the uterus (A) and α-SMA immunohistochemical staining (B) under different concentrations of phloem in Example 2 are shown. Detailed Implementation

[0013] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments, but this should not be construed as limiting the present invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and substance of the invention are within the scope of the present invention. Experimental methods and reagents not specifically described in the embodiments are performed according to conventional conditions in the art.

[0014] The experimental materials used in the following examples are as follows:

[0015] 1.1 Drugs

[0016] Oroxylin A (OA), chemically known as 5,7-dihydroxy-6-methoxy-2-phenyl-4H-1-benzofuran-4-one, has the chemical formula C0. 16 H 12 O5, with a molecular weight of 284.263, was provided by China Pharmaceutical University. It is a pale yellow powder with a purity >99%. Sodium carboxymethyl cellulose (CMC-Na) was used as a solubilizer to prepare a suspension of the pectin and a stock solution of a certain concentration for in vivo experiments.

[0017] 1.2 Mice

[0018] Female BALB / c mice (6-8 weeks old) were purchased from Jiangsu Huachuang Xinno Pharmaceutical Technology Co., Ltd. All mice were housed in an SPF-grade animal facility with a temperature of 20-22℃ and humidity of 40-60%, using a 12-hour light / dark cycle, and with adequate food and water supply.

[0019] According to the inventors' previous research (Jiang Q, Li J, Pan Y, Wang J, Yang J, Shen S) , et al(Stem Cells 2022). An intrauterine adhesion (IUA) model was established using an electric curettage method, i.e., curettage for 3 × 8 s. Thirty-two mice were randomly divided into four groups (n = 8): control group, IUA group, low-dose linolenic acid treatment group, and high-dose linolenic acid treatment group. The control group received only an intraperitoneal injection of the same dose of solvent; the IUA group received an intraperitoneal injection of the same dose of solvent after the electric curettage model was established; the low-dose linolenic acid group received an intraperitoneal injection of 20 mg / kg linolenic acid daily 2 hours after the electric curettage model was established; and the high-dose linolenic acid group received an intraperitoneal injection of 40 mg / kg linolenic acid daily 2 hours after the electric curettage model was established. Mice were sacrificed at 12 hours and 7 days, respectively, and serum and uterine samples were collected to detect the levels of inflammation and fibrosis.

[0020] Example 1

[0021] Thousand-layer paper extract can improve the morphology of the endometrium in mice with intrauterine adhesions and reduce its inflammation level.

[0022] Hematoxylin-eosin staining, or HE staining for short, is used to observe morphological changes in the endometrium. Its principle involves using two dyes—the basic dye hematoxylin and the acidic dye eosin—to act on the cell nucleus and cytoplasm, respectively. This causes the cell's fine structures to change their refractive index through color, thus revealing a clear image of the cells under a light microscope.

[0023] like Figure 1 As shown in Figure A, HE staining of uterine tissue revealed that, compared with the control group, the IUA group mice had incomplete endometrial structure, reduced glandular number, and increased inflammatory cell infiltration. However, after treatment with linalool, the endometrial structure returned to normal, glandular number increased, and inflammatory cell infiltration decreased. Real-time quantitative PCR analysis of the uterus showed that after linalool treatment, the expression of inflammatory factors such as IL-1β, IL-18, and IL-6 was significantly decreased. Figure 1 B). The expression of inflammatory factors such as IL-1β, IL-18, and IL-6 in serum also decreased significantly after treatment with linalool, and the treatment effect of the high-dose group was better than that of the low-dose group. Figure 1 C). This indicates that styrax can improve the morphology of the endometrium in mice with intrauterine adhesions and reduce its inflammation level.

[0024] Example 2

[0025] Thousand-layer paper extract can alleviate the degree of uterine fibrosis in mice with intrauterine adhesions.

[0026] Masson staining, used to reflect the degree of uterine fibrosis, involves mixing two or three anionic dyes to make collagen fibers appear blue and muscle fibers appear red, allowing for observation of the collagen structure in the tissue. α-SMA is a marker of smooth muscle cells and is currently also considered a marker of myofibroblasts; immunohistochemical staining of uterine tissue with α-SMA can also reflect the degree of uterine fibrosis.

[0027] like Figure 2 As shown, Masson staining and α-SMA immunohistochemical staining of uterine tissue revealed that, compared with the control group, the endometrial stroma of mice in the IUA group was replaced by collagen. Figure 2 A), α-SMA + The number of myofibroblasts increased significantly. Figure 2 B), but after treatment with hymenoplasty, the amount of collagen fibers and α-SMA in the endometrium... + The number of myofibroblasts decreased significantly in all groups, and the high-dose group showed better therapeutic effects than the low-dose group. This indicates that linalool can alleviate the degree of uterine fibrosis in mice with intrauterine adhesions.

Claims

1. Application of Thousand-Layer Paper Pulp in the Preparation of Drugs for the Treatment of Intrauterine Adhesions.

2. The application according to claim 1, characterized in that, The thousand-layer paper extract improves the morphology of endometrial adhesions, reduces the expression of inflammatory factors IL-1β, IL-18, and IL-6 in uterine tissue, and decreases the expression of inflammatory factors IL-1β, IL-18, and IL-6 in serum.

3. The application according to claim 1, characterized in that, The layered paper reduces the degree of fibrosis in the uterine cavity due to adhesions.

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