Application of indole-3-formaldehyde in preparation of enteric nerve protection medicine

Abstract

The invention belongs to the field of medicines, and provides application of indole-3-formaldehyde in preparation of an enteric nerve protection medicine. A loperamide-induced C57BL / 6J mouse constipation model is adopted, it is found that indole-3-carboxaldehyde can accelerate the small intestine propulsion rate of a model mouse, increase the water content of excrement and the excretion number of excrement particles in unit time and improve the intestinal motility of the model mouse, and HUC / DmRNA expression of mouse colon tissue treated by indole-3-carboxaldehyde is remarkably increased compared with a model group; protein expression of HUC / D and HSP90AA1 is remarkably increased compared with that of a model group, immunohistochemistry and immunofluorescence show that the number of HUC / D dyed neurons and fluorescence intensity of colon tissue treated by indole-3-formaldehyde are remarkably increased compared with that of the model group, and therefore, the indole-3-formaldehyde can promote intestinal motility by protecting HUC / D intestinal neuronal cells.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and specifically relates to the application of indole-3-carboxaldehyde in the preparation of enteric nerve protective drugs. Background Art

[0002] Indole-3-carboxaldehyde is a tryptophan metabolite derived from microorganisms, also known as β-indoleacetaldehyde, 3-indolecarboxaldehyde, 3-formylindole, NSC 10118, indole-3-acetaldehyde, with a molecular formula of C9H7NO. Previous studies have shown that indole-3-carboxaldehyde has anti-inflammatory effects, protects the intestinal epithelial barrier, and improves metabolic complications caused by a high-fat diet.

[0003] It is believed in the prior art that the administration of indole-3-carboxaldehyde can accelerate the intestinal peristalsis of zebrafish and improve their intestinal motility.

[0004] The enteric nervous system originates from neural crest stem cells and is composed of enteric neurons and glial cells. It is divided into the myenteric plexus and the submucosal plexus. It can express a variety of different neurotransmitters and neuropeptides, participate in the initiation and control of the contractile function of intestinal smooth muscle, and affect intestinal motility.

[0005] Various external and internal factors faced by the gastrointestinal tract, such as genetic factors, exposure to toxic substances during development, and changes in dietary structure, may affect the function of individual neuronal subtypes or the entire enteric nervous system, thereby causing changes in intestinal motility. In patients with congenital megacolon, the absence of intermuscular ganglion cells in the colon wall can be seen, causing spasmodic contraction of the diseased colon, secondary expansion of the proximal intestinal tract to form megacolon, and ultimately obstruction of intestinal contents discharge.

[0006] Mice fed a Western diet showed increased pyroptosis of nNOS enteric neurons in the colonic myenteric compartment, delayed colonic transit, and impaired colonic relaxation responses induced by electrical stimulation in isolated colonic tissue. Loss of intestinal nNOS neurons was observed in a diabetic rat model, resulting in reduced small intestinal propulsion rate and delayed gastric emptying. In addition, a significant reduction in intestinal Chat neurons in starved rats can lead to a significant decrease in intestinal motility. The enteric nervous system plays an important role in intestinal motility, and damage to intestinal nerves inhibits intestinal motility.

[0007] Existing technology believes that some probiotics, such as Lactobacillus paracasei, can increase the number of intestinal glial cells to repair intestinal nerves; however, there are few studies on intestinal neuroprotective drugs.

[0008] However, the prior art does not propose whether the improvement of indole-3-carboxaldehyde on intestinal motility works through enteric nerves. Summary of the invention

[0009] In order to solve the above technical problems, the present invention provides the use of indole-3-carboxaldehyde in the preparation of enteric nerve protective drugs to solve the problem that the prior art has not proposed whether the improvement of indole-3-carboxaldehyde on intestinal motility works through enteric nerves.

[0010] Application of indole-3-carboxaldehyde in the preparation of enteric neuroprotective drugs.

[0011] Preferably, the drug is used to treat one or more of constipation, constipation-type irritable bowel syndrome, and functional dyspepsia.

[0012] Preferably, the pharmaceutically active ingredient includes: indole-3-carboxaldehyde or a pharmaceutically acceptable salt compound, hydrate, solvate, enantiomer, stereoisomer or tautomer thereof.

[0013] Preferably, the drug can be administered orally, by gavage, enema, subcutaneous injection, or skin application.

[0014] Preferably, indole-3-carboxaldehyde protects intestinal HUC / D neuronal cells by activating the HSP90AA1 pathway.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] 1. The present invention adopts a C57BL / 6J mouse constipation model induced by loperamide, and finds that indole-3-formaldehyde can accelerate the small intestinal propulsion rate of the model mice, increase the stool water content and the number of stool particles discharged per unit time, and improve the intestinal motility of the model mice. Moreover, after indole-3-formaldehyde treatment, the HUC / D mRNA expression of the mouse colon tissue is significantly increased compared with the model group, and the protein expression of HUC / D and HSP90AA1 is significantly increased compared with the model group. Immunohistochemistry and immunofluorescence show that the number and fluorescence intensity of HUC / D stained neurons in the colon tissue are significantly increased compared with the model group after indole-3-formaldehyde treatment. Therefore, indole-3-formaldehyde can promote intestinal motility by protecting HUC / D enteric neuron cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a comparison chart of fecal water content in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group, among which: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0018] Figure 2 It is a comparison chart of the number of fecal particles per unit time in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group, where: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0019] Figure 3This is a comparison chart of the small intestinal propulsion rate among the normal control group, loperamide modeling group and indole-3-carboxaldehyde group, where: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0020] Figure 4 The expression of HUC / D in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group (immunohistochemical staining), where: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0021] Figure 5 The expression of HUC / D in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group (immunofluorescence staining), where: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0022] Figure 6 The representative diagrams of HUC / D expression in single ganglia in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group (colonic nerve flat mount, immunofluorescence staining), among which: Controll is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0023] Figure 7 It is a comparison chart of HUC / D mRNA expression in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group, where: Control is the normal control group, IALD is the loperamide + indole-3-carboxaldehyde group;

[0024] Figure 8 The figure is a comparison chart of the expression and relative expression of HUC / D and HSP90AA1 in the normal control group, loperamide modeling group and indole-3-carboxaldehyde group, where: Control is the normal control group, and IALD is the loperamide + indole-3-carboxaldehyde group. DETAILED DESCRIPTION

[0025] The following embodiments of the present invention are described in further detail in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0026] Experimental animals and animal model preparation: Female C57BL / 6J mice (purchased from Beijing Huafukang Biotechnology Co., Ltd.) were selected, with an age of 6-8 weeks and a body weight of 18-20 g.

[0027] Embodiment 1:

[0028] Grouping and treatment: (1) Mice were divided into control group (Control), loperamide group, and loperamide + indole-3-carboxaldehyde group (IALD). Mice in loperamide group and IALD group were gavaged with 60 mg / kg loperamide for 2 weeks. Mice in IALD group were gavaged with 50 mg / kg indole-3-carboxaldehyde daily while gavaged with loperamide. Intestinal motility was tested after two weeks. The specific modeling scheme is shown in Table 1.

[0029] Table 1 Modeling scheme

[0030]

[0031] Example 2: The promoting effect of indole-3-carboxaldehyde on intestinal motility.

[0032] (2) Detect the moisture content of mouse feces. After 2 weeks of gavage, each group of mice was placed in a separate cage, and the bedding was replaced with new ones. The mice were fed and watered normally. After 2 hours, the feces in each cage were collected using a 1.5 ml EP tube (those stained with urine were discarded). The weight of the fresh feces was then weighed and recorded as the wet weight. The feces were dried and weighed again and recorded as the dry weight. Moisture content = (wet weight - dry weight) / wet weight × 100%. The results are shown in the attached Figure 1 As shown, indole-3-carboxaldehyde increased the water content of feces in the loperamide modeling group mice.

[0033] (3) Count the number of fecal particles per unit time of the mice. As shown in step 3, at 0.5 hours, use a 1.5 ml EP tube to collect all feces in each cage and count the number of feces. Figure 2 As shown, indole-3-carboxaldehyde increased the number of stool particles per unit time in the loperamide modeling group mice.

[0034] (4) Detection of the small intestinal propulsion rate of mice. After the model was established, each group of mice was gavaged with Evans blue solution. After 15 minutes, the mice were decapitated and the small intestine was removed to measure the propulsion distance of the Evans blue solution. Small intestinal propulsion rate = Evans blue solution propulsion distance / small intestinal length × 100%. Figure 2 As shown, indole-3-carboxaldehyde increased the small intestinal propulsion rate of loperamide modeling mice.

[0035] Example 3: Protective effect of indole-3-carboxaldehyde on intestinal nerves in mice.

[0036] (5) To evaluate the protective effect of indole-3-formaldehyde on HUC / D enteric nerves. The number of HUC / D nerves in the myenteric plexus in the control group, loperamide group, and indole-3-formaldehyde group was observed by immunohistochemistry, immunofluorescence, and enteric nerve flat mount. Figure 4-6 As shown, indole-3-carboxaldehyde has a protective effect on enteric nerves.

[0037] Example 4: Exploration of the mechanism of the protective effect of indole-3-carboxaldehyde on mouse enteric nerves.

[0038] (6) After treatment with indole-3-formaldehyde, the expression of HUC / D mRNA level in mice in the IALD group was significantly increased, such as Figure 7 As shown in Figure 2, the results of Western blot indicated that the expression of HUC / D and HSP90AA1 proteins in the IALD group was significantly increased. Figure 8 shown.

[0039] The embodiments of the present invention are provided for the purpose of illustration and description. Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations of the present invention. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. Application of indole-3-carboxaldehyde in the preparation of enteric neuroprotective drugs.

2. The use according to claim 1, characterized in that: The medicine is used to treat one or more of constipation, constipation-type irritable bowel syndrome, and functional dyspepsia.

3. The use according to claim 1, characterized in that: The active pharmaceutical ingredient comprises: indole-3-carboxaldehyde or a pharmaceutically acceptable salt compound, hydrate, solvate, enantiomer, stereoisomer or tautomer thereof.

4. The use according to claim 1, characterized in that: The drug can be administered orally, by gavage, enema, subcutaneous injection, or skin application.

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