Application of Piezo1 inhibitor in preparation of medicine for preventing and treating esophageal stenosis

Abstract

The invention discloses an application of a Piezo1 inhibitor in preparation of a medicine for treating esophageal stenosis. It is found for the first time that esophageal endothelial cells of esophageal stenosis patients or models have Piezo1 high expression, the Piezo1 inhibitor can effectively reduce inflammatory cell infiltration under esophageal mucosa, reduce collagenous fiber deposition under the esophageal mucosa and inhibit the endothelial cells from being converted into fibroblasts, and then fibrosis is inhibited, so that esophageal stenosis is effectively reversed, prevented and treated.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to the use of Piezo1 inhibitors in the preparation of medicaments for the prevention and treatment of esophageal stricture. Background Technology

[0002] Esophageal stricture (ES) refers to obstruction of the esophageal passage, leading to a series of symptoms, primarily dysphagia. In children, esophageal stricture is a common acquired esophageal disease, mainly caused by chemical burns to the esophagus. Its core pathology involves the massive deposition of esophageal collagen fibers, ultimately forming fibrotic esophageal stricture, resulting in dysphagia, feeding difficulties, and even malnutrition. Treatment primarily involves endoscopic balloon dilation, but the recurrence rate is high, posing a significant threat to the child's growth, development, and overall health.

[0003] The exact reasons for the massive deposition of esophageal collagen fibers during the pathogenesis of endometriosis (ES) remain unclear, but vascular endothelial cells play a crucial role in regulating collagen synthesis. Under chronic inflammatory stimulation, vascular endothelial cells acquire stromal cell function and transform into mesenchymal cells by expressing alpha-smooth muscle actin (α-SMA) and vimentin. This process is called endothelial-to-mesenchymal transition (EndMT) and plays an important role in the fibrotic process of various diseases.

[0004] In gastrointestinal fibrosis, repeated and chronic injury (such as chemical burns and inflammatory stimulation) is accompanied by the deposition of large amounts of extracellular matrix (ECM), tissue remodeling, and subsequent luminal narrowing. Elastic modulus, as a key mechanical parameter for measuring tissue stiffness (measured in Pascals, Pa or kilopascals, kPa), quantifies the mechanical properties of the gastrointestinal wall. The elastic modulus of healthy gastrointestinal tissue is typically 2 kPa, while the stiffness of fibrotic gastrointestinal tissue can reach 8-28 kPa, tens of times higher than normal tissue. However, how endothelial cells sense changes in tissue stiffness and regulate their pro-fibrotic functional phenotype in the context of esophageal fibrosis remains a crucial scientific question that needs to be elucidated.

[0005] Currently, there is an urgent need for innovative drugs in the clinical treatment of esophageal stricture that can reverse fibrosis and restore the normal structure of the esophagus. Although treatment strategies based on regulating fibroblast activation or the immune microenvironment have shown great promise, no specific drugs for esophageal fibrosis have been approved for use to date. Summary of the Invention

[0006] This invention aims to at least solve one of the aforementioned technical problems existing in the prior art. Therefore, the object of this invention is to provide the application of Piezo1 inhibitors in the preparation of drugs for treating esophageal stricture. This invention is the first to discover that esophageal endothelial cells in patients or models of esophageal stricture have high expression of Piezo1, and that Piezo1 inhibitors can effectively reduce the infiltration of inflammatory cells in the esophageal submucosa, reduce collagen fiber deposition in the esophageal submucosa, inhibit the transformation of endothelial cells into fibroblasts, and thus inhibit fibrosis, thereby effectively reversing, preventing, and treating esophageal stricture.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A first aspect of the invention provides the use of a Piezo1 inhibitor in the preparation of a medicament for the prevention and / or treatment of esophageal stricture.

[0008] In some embodiments of the present invention, the Piezo1 inhibitor includes at least one of the following: (a1) Substances that reduce the content of Piezo1 protein; (a2) Substances that inhibit the expression of the Piezo1 gene; (a3) Substances that silence the Piezo1 gene; (a4) Substances that knock out the Piezo1 gene; (a5) Substances that inhibit the activity of Piezo1 protein.

[0009] In some embodiments of the present invention, the Piezo1 inhibitor comprises at least one of nucleic acid molecules, protein molecules, and small molecule compounds.

[0010] In some embodiments of the present invention, the nucleic acid molecule includes at least one of microRNA, siRNA, shRNA, dsRNA, sgRNA, and antisense oligonucleotides.

[0011] In this invention, nucleic acid molecules inhibit the expression of Piezo1 protein through "gene silencing" or "targeted degradation of mRNA." By interfering with the transcription or translation processes in the central dogma, Piezo1 protein production is prevented, thereby achieving Piezo1 inhibition. For example, when antisense oligonucleotides bind to mRNA, they form a DNA-RNA heteroduplex, thereby recruiting the intracellular RNase H1 enzyme. This enzyme recognizes this structure and cleaves and degrades the mRNA, preventing it from being translated into protein. Therefore, antisense oligonucleotides that are perfectly complementary to specific regions of Piezo1 mRNA can be designed to cause mRNA degradation through the aforementioned mechanism (mainly the RNase H1 pathway). After siRNA is introduced into the cell, it can bind to the RISC protein complex (RNA-induced silencing complex, RISC). The RISC then unwinds the siRNA double helix, retaining its guide strand. The guide strand finds a perfectly matched Piezo1 mRNA through base complementarity pairing. Subsequently, the AGO2 protein in the RISC (which has "slicer" activity) directly cleaves and degrades the target mRNA. shRNA sequences are typically integrated into the cell's genome via viral vectors (such as lentiviruses and adeno-associated viruses) to achieve long-term, stable expression. Cellular enzymes (Drosha and Dicer) cleave the hairpin structure of the shRNA, generating functional siRNA. The siRNA then guides the degradation of Piezo1 mRNA through the aforementioned mechanism. miRNAs are endogenous non-coding small RNAs that typically target multiple mRNAs through imperfect complementary pairing. After transcription, miRNA genes undergo a series of processing steps to form mature single-stranded miRNAs. The miRNA's "seed sequence" (usually the nucleotide sequence from position 2 to position 8 at the 5' end of the mature miRNA molecule) binds to the 3' untranslated region of the target mRNA, thereby inhibiting mRNA translation. Therefore, miRNA mimics can be designed to mimic the function of endogenous miRNAs. When introduced into cells, these mimics can target and inhibit the translation of Piezo1 mRNA. sgRNA is a chimeric RNA in which a portion of the sequence is designed to be complementary to a specific DNA site of the Piezo1 gene and to form a complex with the Cas9 nuclease. The sgRNA "navigates" the Cas9 enzyme to the Piezo1 gene on the genome, where Cas9 will cause a DNA double-strand break, thereby inactivating the Piezo1 gene.

[0012] In some embodiments of the present invention, the protein molecule includes a polypeptide.

[0013] In some embodiments of the present invention, the polypeptide includes a Piezo1-specific antibody and other non-antibody peptides capable of inhibiting Piezo1 activity.

[0014] As is known to those skilled in the art, Piezo1 is the first member of a family of mechanogated cation channels in mammals. As a mechanosensor, this channel senses changes in cell membrane mechanical forces, triggering calcium ion influx and converting mechanical signals into bioelectrical signals, thus participating in physiological processes such as angiogenesis, erythrocyte volume regulation, and blood pressure homeostasis.

[0015] In some embodiments of the present invention, the Piezo1-specific antibody is a Piezo1 neutralizing antibody or a Piezo1 function-blocking antibody. For example, the antibody can bind to a specific epitope of the Piezo1 protein with high affinity through its antigen-binding fragment (Fab region), thereby inhibiting pore opening (e.g., the antibody binds to or near the entrance of an ion channel, directly blocking the entry and exit of ions, thereby inhibiting current; or by binding to the Piezo1 mechanosensing domain, inhibiting channel opening) or interfering with gating motion (e.g., Piezo1 undergoes a conformational change after sensing membrane tension, thereby opening the pore; the antibody binds to the key mechanosensing domain or gating domain responsible for this conformational change, which can prevent the channel from opening).

[0016] In some embodiments of the present invention, the polypeptide includes GsMTx4.

[0017] GsMTx4 is a polypeptide toxin extracted from the venom of the tarantula (Grammostola spatulata). It can indirectly inhibit the sensitivity of Piezo1 channels to mechanical stimulation by binding to the phospholipids on the outer layer of the cell membrane and altering the membrane's mechanical properties.

[0018] In some embodiments of the present invention, the small molecule compounds include, but are not limited to, Dooku1, quercetin, isoquercetin, jatrorrhizine, and gadolinium (Gd). 3+ ), ruthenium red, fritillary glycoside A, artemisinin.

[0019] In some embodiments of the present invention, the medicament includes pharmaceutically acceptable excipients.

[0020] In one embodiment of the present invention, the pharmaceutically acceptable excipients include at least one of the following: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, release inhibitors, and carriers.

[0021] In some embodiments of the present invention, the dosage form of the drug includes a gastrointestinal dosage form or a non-gastrointestinal dosage form.

[0022] In some embodiments of the present invention, the gastrointestinal dosage form includes at least one of powder, tablet, granule, capsule, sustained-release, solution, dry suspension, effervescent tablet, emulsion, suspension, syrup, drops, and chewable tablet.

[0023] In some embodiments of the present invention, the non-gastrointestinal dosage form includes at least one of injection dosage form, respiratory dosage form, skin dosage form, mucosal dosage form, and cavity dosage form.

[0024] In some embodiments of the present invention, the route of administration of the drug includes at least one of intravenous injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, oral administration, sublingual administration, nasal administration, nebulized administration, or transdermal administration.

[0025] In some embodiments of the present invention, the dosage range of the drug is 0.05-0.20 mg / kg / d.

[0026] In some embodiments of the present invention, the subjects of the drug include infants, children, adolescents, and adults.

[0027] In this invention, "esophageal stricture" refers to obstruction of the esophageal passage, leading to a series of symptoms centered on dysphagia. Common causes of esophageal stricture include, but are not limited to, reflux esophagitis (continuous irritation and damage to the mucosa of the lower esophagus by refluxed gastric acid and bile), chemical esophageal injury (esophageal damage caused by the adhesion of corrosive chemicals such as strong acids and alkalis or irritating drugs to the esophageal wall), mechanical esophageal injury (e.g., iatrogenic injury caused by endoscopy or treatment, surgery or radiation therapy, or foreign body injury), and eosinophilic esophagitis (allergens (such as certain foods) trigger the infiltration of a large number of eosinophils into the esophageal wall, leading to chronic inflammation, ... Edema and fibrosis eventually cause the esophagus to lose elasticity and form multiple annular strictures; infectious esophagitis (infection caused by pathogenic microorganisms and extensive ulceration of the esophageal mucosa, usually occurring in people with weakened immune function); neoplastic strictures (esophageal cancer (squamous or adenocarcinoma) or gastric cardia cancer invading the esophagus, with tumor tissue growing into the lumen and directly obstructing the esophageal lumen); and congenital and systemic diseases (including congenital esophageal strictures, esophageal webs and esophageal rings, and esophageal manifestations of systemic diseases such as scleroderma, Crohn's disease, Behcet's disease, etc.).

[0028] In some embodiments of the present invention, the esophageal stricture includes esophageal stricture caused by chemical damage, esophageal stricture caused by mechanical damage, and esophageal stricture caused by reflux esophagitis.

[0029] The beneficial effects of this invention are: This invention is the first to discover that esophageal endothelial cells in patients or models of esophageal stricture have high expression of Piezo1. Piezo1 inhibitors can effectively reduce the infiltration of inflammatory cells in the esophageal submucosa, reduce the deposition of collagen fibers in the esophageal submucosa, inhibit the transformation of endothelial cells into fibroblasts, and thus inhibit fibrosis, thereby effectively reversing, preventing and treating esophageal stricture. Attached Figure Description

[0030] Figure 1 The expression of Piezo1 in esophageal endothelial cells of patients with esophageal stricture is shown. A and B are the Masson staining results (A) and immunofluorescence staining results (B) of esophageal tissue samples from patients with esophageal stricture and control tissue samples, respectively.

[0031] Figure 2 The image shows the expression of Piezo1 in esophageal endothelial cells of a rat esophageal stricture model. A and D represent Masson staining results (A), gene expression results (B), protein expression results (C), and immunofluorescence staining results (D) of esophageal tissues from rats in the esophageal stricture group and the control group, respectively.

[0032] Figure 3 The effect of the Piezo1 inhibitor GsMTx4 on a rat model of esophageal stricture is shown in Figures A, C, and D, respectively, representing the changes in body weight (A), hematoxylin-eosin and masson staining results (B), and immunofluorescence staining results (C) of the three groups of rats (control group, NaOH+ solvent model group, and NaOH+GsMTx4 model group). Detailed Implementation

[0033] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0034] Furthermore, to better illustrate the present invention, numerous specific details are set forth in the following detailed embodiments. Those skilled in the art should understand that the present invention can be practiced without certain specific details. In some embodiments, materials, methods, and means well known to those skilled in the art are not described in detail in order to highlight the spirit of the invention.

[0035] Example 1: Expression of Piezo1 in esophageal endothelial cells of patients with esophageal stricture Tissue specimens from patients undergoing endoscopic balloon dilation for corrosive esophageal stricture and healthy control tissue samples (from the Women and Children's Medical Center Affiliated to Guangzhou Medical University) were collected. Esophageal collagen was detected by Masson staining, and Piezo1 expression in esophageal endothelial cells was detected by immunofluorescence using vascular endothelial cell markers (CD31) and Piezo1 antibody. The specific steps are as follows: Masson staining: (1) Dewax sections to distilled water according to routine procedure, following the instructions of the Masson's Trichrome Staining Solution Kit (solarbio, G1345).

[0036] (2) Prepare Weigert iron hematoxylin staining solution by mixing reagents A1 and A2 in a 1:1 ratio according to the kit instructions. Add a certain amount of staining solution to cover the slide and stain for 10 minutes.

[0037] (3) Use distilled water to wash away excess staining solution, add acidic differentiation solution for 10 seconds, and then react with distilled water for 30 seconds.

[0038] (4) Add Masson blue solution for 5 minutes to turn blue, then add distilled water for 30 seconds.

[0039] (5) Add Ponceau S and Fuchsia staining solution and stain for 10 minutes.

[0040] (6) Prepare a weak acid working solution by mixing distilled water and weak acid solution in a 2:1 ratio, and add the weak acid working solution dropwise for 30 seconds.

[0041] (7) Discard the excess liquid, add phosphomolybdic acid solution for 1 minute, and add weak acid working solution for 30 seconds.

[0042] (8) Discard the excess liquid, add aniline blue staining solution to stain for 1 minute, and add weak acid working solution to react for 30 seconds.

[0043] (9) Dehydration and clearing steps (immerse in 75% alcohol, 85% alcohol, 95% alcohol, and 100% alcohol (I) for 2 seconds each, and immerse in 100% alcohol (II) for 1 minute; clearing: clear with xylene twice (1 minute each time)).

[0044] (10) Mount the slide with neutral resin to avoid air bubbles, and observe it under a microscope after the resin dries.

[0045] Immunofluorescence staining: (1) Dewax the sections to distilled water using standard procedures.

[0046] (2) Repair: Prepare 1× sodium citrate antigen repair solution (Biosharp, BL151A), put the slide into the antigen repair solution, heat it in the microwave oven on high for 15 minutes, and discard the antigen repair solution after the slide has cooled to room temperature. Wash with PBS solution 3 times, 5 minutes each time.

[0047] (3) Permeabilization: Add 0.3% Triton-100 (Beyotime, ST1723) to the slice to cover the tissue, and incubate at room temperature for 15 minutes.

[0048] (4) Blocking: Add an appropriate amount of blocking goat serum (Biosharp, BL1092B) to cover the tissue, incubate at room temperature for 1 hour, and then discard the liquid.

[0049] (5) Primary antibody incubation: Add appropriate amounts of diluted CD31 antibody (Proteintech, 11265-1, 1:100) and Piezo1 antibody (Huabio, M1005-2, 1:200) to cover the esophageal tissue and incubate overnight at 4°C.

[0050] (6) Secondary antibody incubation: The next day, place the slide in PBS solution and shake it three times for 5 minutes each time; add the corresponding 594-Goat Anti-Rabbit Secondary Antibody (Proteintech, RGAR004, 1:200) or 488-Goat Anti-Mouse Secondary Antibody (Proteintech, RGAM002, 1:200) secondary antibody incubation solution according to the species of primary antibody, and incubate at room temperature for 1 hour. After incubation, place the slide in PBS solution and shake it three times for 5 minutes each time.

[0051] (7) Mounting: Shake off the liquid on the slide, add an appropriate amount of mounting medium containing DAPI (Beyotime, P0131) to mount the slide, and observe and acquire images under a fluorescence microscope.

[0052] The results are as follows Figure 1 As shown, the experimental results indicate that, compared to the control group, patients with esophageal stricture exhibit disordered esophageal structure and increased collagen deposition in the tissue. Figure 1 In A), endothelial cells showed increased Piezo1 expression ( Figure 1 (B in the middle).

[0053] Example 2: Expression of Piezo1 in esophageal endothelial cells of a rat esophageal stricture model Experimental subjects: 10 male SD rats, weighing approximately 300 g, were randomly divided into groups of 5 rats each.

[0054] Experimental grouping: There were two groups: control group and esophageal stricture group.

[0055] Experimental steps: Rats were randomly divided into a control group (n=5) and an esophageal stricture group (n=5). Rats were anesthetized with 2.5% tribromoethanol (6 mL / kg). After anesthesia took effect, for the esophageal stricture group, 0.1 mL of 20% NaOH solution was slowly instilled into the esophagus through a double-lumen gavage tube. After maintaining this for 1 minute, the NaOH solution was carefully withdrawn, and the esophagus was immediately flushed repeatedly with distilled water. After flushing, the rats were deprived of food and water for 24 hours. For the control group, under the same anesthesia, an equal volume of distilled water was instilled into the esophagus through a double-lumen gavage tube. The rats were also deprived of food and water for 24 hours after the procedure. The rats' feeding status, weight changes, and stool shape and color were continuously observed and recorded in detail. On day 28, all experimental animals were euthanized and their tissues were collected. Esophageal tissue samples were collected. RNA was extracted from one portion for quantitative real-time polymerase chain reaction (qPCR), and protein was extracted from another portion for Western blot analysis. The remaining tissue was embedded and sectioned for Masson staining to observe morphological changes and determine the success of the stricture model. After confirming successful model establishment, immunofluorescence staining was performed to observe Piezo1 expression in endothelial cells. CD31 is a marker for esophageal endothelial cells. The steps for Masson staining and immunofluorescence staining are described in Example 1. GAPDH gene expression was used as an internal control. The primer sequences used in the qPCR experiment are shown in Table 1 below. The antibody information used in the Western blot experiment is as follows: the primary antibodies were Piezo1 antibody (Huabio, M1005-2, 1:1000) and α-tubulin (Rakan, RM2007, 1:5000), and the secondary antibodies were HRP-Goat-anti-Rabbit antibody (Jackson, 111-035-003, 1:2000) and HRP-Goat-anti-Mouse antibody (Jackson, 115-035-003, 1:2000).

[0056] Table 1. Primer sequences

[0057] Experimental results The results of masson staining showed that, compared with the control group, the esophageal stricture group rats had disordered submucosal structures of the esophagus and significantly increased blue collagen deposition. Figure 2 In addition, compared with the control group, the expression levels of Piezo1 mRNA and protein in the esophageal tissue of rats in the esophageal stricture group were significantly increased (A). Figure 2 (B and C in the text). Immunofluorescence results also showed that, compared with the control group, the expression of Piezo1 in endothelial cells of esophageal tissue in rats with esophageal stricture was significantly increased (B and C in the text). Figure 2 (D in the middle).

[0058] The above experimental results indicate that the expression of Piezo1 in the esophagus of rats with esophageal stricture is consistent with that in the esophageal tissue of patients with esophageal stricture; that is, in the case of esophageal stricture, the expression of Piezo1 in the endothelial cells of both rat models and patients' esophageal tissues is significantly increased.

[0059] Example 3: Effects of Piezo1 inhibitor GsMTx4 on a rat model of esophageal stricture Experimental subjects: 15 male SD rats, weighing 300 g, were randomly divided into groups of 5 rats each.

[0060] Experimental grouping: There were 3 groups in total: control group, NaOH + solvent treatment group and NaOH + GsMTx4 treatment group, with 5 animals in each group.

[0061] Experimental steps: The model construction method for the control group was the same as in Example 2. The model construction methods for the NaOH + solvent treatment group and the NaOH + GsMTx4 treatment group are as follows: The method for constructing a rat model of esophageal stricture using NaOH was the same as in Example 2. On day 7 after model construction, the rats with esophageal stricture were randomly divided into a NaOH + solvent treatment group and a NaOH + GsMTx4 treatment group. The solvent treatment group and the GsMTx4 treatment group received intraperitoneal injections of either solvent (10% DMSO + 90% corn oil) or GsMTx4 (1 mg / kg) (MCE, HY-P1410) on days 14, 21, 28, and 35, respectively. The rats' feeding status, weight changes, and stool morphology and color were continuously observed and recorded in detail. On day 42, all experimental animals were euthanized and esophageal tissue samples were collected. These samples were embedded and sectioned, and H&E staining was used to observe morphological changes. Masson staining was used to detect collagen fiber deposition, and immunofluorescence staining was used to label specific proteins to clarify their distribution in the tissue. H&E staining was performed according to standard experimental procedures in the art. The steps for Masson staining and immunofluorescence staining are described in Example 1. The information of the α-SMA antibody used in immunofluorescence staining is as follows: Proteintech, 14395-1-AP, dilution ratio 1:1000.

[0062] Experimental results Changes in rat body weight as follows Figure 3 As shown in A, the results indicate that the body weight of rats in the esophageal stricture group increased significantly after treatment with the Piezo1 inhibitor GsMTx4.

[0063] The effect of GsMTx4 on pathological damage of rat esophagus is as follows: Figure 3 As shown in B and C, the results indicate that after treatment with the Piezo1 inhibitor GsMTx4, H&E staining showed that GsMTx4 reduced the infiltration of inflammatory cells in the esophageal submucosa, and Masson staining showed that the blue collagen fibers in the esophageal submucosa of the esophageal stricture group were significantly reduced after GsMTx4 treatment. Figure 3 (B in the text); Immunofluorescence staining results also indicated that GsMTx4 could reduce the colocalization of esophageal endothelial cell marker (CD31) and fibroblast marker (a-SMA) in the esophageal stricture model, suggesting that GsMTx4 can inhibit the transformation of endothelial cells into fibroblasts, thereby inhibiting fibrosis. Figure 3 (C in the middle).

[0064] The above experimental results indicate that the Piezo1 inhibitor GsMTx4 can improve the body weight of rats with esophageal stricture and reduce esophageal collagen deposition, thus effectively treating esophageal stricture.

[0065] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The use of Piezo1 inhibitors in the preparation of drugs for the prevention and / or treatment of esophageal stricture.

2. The application according to claim 1, characterized in that, The Piezo1 inhibitor includes at least one of the following: (a1) Substances that reduce the content of Piezo1 protein; (a2) Substances that inhibit the expression of the Piezo1 gene; (a3) Substances that silence the Piezo1 gene; (a4) Substances that knock out the Piezo1 gene; (a5) Substances that inhibit the activity of Piezo1 protein.

3. The application according to claim 1, characterized in that, Piezo1 inhibitors include at least one of nucleic acid molecules, protein molecules, and small molecule compounds.

4. The application according to claim 3, characterized in that, The nucleic acid molecules include at least one of microRNA, siRNA, shRNA, dsRNA, sgRNA, and antisense oligonucleotides.

5. The application according to claim 3, characterized in that, The protein molecule includes a polypeptide, and the polypeptide includes a Piezo1-specific antibody.

6. The application according to claim 5, characterized in that, The polypeptide includes GsMTx4.

7. The application according to claim 3, characterized in that, The small molecule compounds include Dooku1, quercetin, isoquercetin, jatrorrhizine, and gadolinium (Gd). 3+ At least one of the following: ruthenium red, fritillary glycoside A, and artemisinin.

8. The application according to claim 1, characterized in that, The drug comprises pharmaceutically acceptable excipients, which include at least one of the following: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, integrators, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and anti-flocculators, filter aids, release inhibitors, and carriers.

9. The application according to claim 1, characterized in that, The dosage form of the drug includes gastrointestinal dosage forms or non-gastrointestinal dosage forms; the gastrointestinal dosage forms include at least one of powder, tablet, granule, capsule, sustained-release, solution, dry suspension, effervescent tablet, emulsion, suspension, syrup, drops, and chewable tablet; the non-gastrointestinal dosage forms include at least one of injection dosage forms, respiratory dosage forms, skin dosage forms, mucosal dosage forms, and cavity dosage forms.

10. The application according to claim 1, characterized in that, The esophageal strictures include those caused by chemical damage, mechanical damage, and reflux esophagitis.

Images