CN114736962A

CN114736962A - Application of inhibitor of circDHTKD1 in preparation of medicine for regulating and controlling airway epithelial inflammation

Info

Publication number: CN114736962A
Application number: CN202210566976.XA
Authority: CN
Inventors: 钱粉红; 何山川; 庄琼馨; 杨贤苗; 严梦楠; 陈欣欣; 赵思婷; 曹宇; 刘雨雪
Original assignee: Affiliated Hospital of Jiangsu University
Current assignee: Affiliated Hospital of Jiangsu University
Priority date: 2022-05-24
Filing date: 2022-05-24
Publication date: 2022-07-12
Anticipated expiration: 2042-05-24
Also published as: CN114736962B

Abstract

The invention discloses an application of an inhibitor of circDHTKD1 in preparation of a medicine for regulating airway epithelial inflammation, and belongs to the field of biological medicines. The invention provides an application of a reagent for detecting the expression level of circDHTKD1 in the preparation of a kit for diagnosing and/or detecting airway epithelial inflammation; also provides an application of the reagent for detecting the expression level of circDHTKD1 in preparing a kit for diagnosing and/or detecting asthma. The invention provides an application of an inhibitor of circDHTKD1 in preparation of a medicine for regulating airway epithelial inflammation and an application of an inhibitor of circDHTKD1 in preparation of a medicine for treating asthma. The invention verifies the action mechanism of circDHTKD1, provides a potential target spot for regulating and controlling human airway epithelial inflammation injury clinically, and provides a theoretical basis for the research of related medicaments.

Description

Application of inhibitor of circDHTKD1 in preparation of medicine for regulating and controlling airway epithelial inflammation

Technical Field

The invention relates to an application of an inhibitor of circDHTKD1 in preparation of a medicine for regulating airway epithelial inflammation, and belongs to the field of biological medicines.

Background

Bronchial asthma is a chronic airway disease characterized by airway inflammation, airway hyperresponsiveness, and airway remodeling, with airway epithelium being an important participant in airway inflammation and airway innate immunity. Circular RNA (circular RNA) is a closed circular non-coding RNA, and more studies have confirmed that circular RNA is widely involved in the pathophysiology of various diseases.

However, the mechanism by which circular RNA participates in the regulation of airway epithelial inflammatory responses is currently unknown. Therefore, the deep research on the action mechanism of the circRNA for regulating airway inflammation can provide a new strategy for the diagnosis and treatment of asthma, and has important significance.

Disclosure of Invention

The invention aims to discuss the application of an inhibitor of circDHTKD1 in preparing a medicine for regulating airway epithelial inflammation, the inflammatory effect of LPS on human bronchial epithelial cells (BEAS-2B) and the regulating effect of circular RNA (circDHTKD1) in the medicine.

In order to realize the technical scheme, the invention provides the following technical scheme:

the first technical scheme is as follows: application of an agent for detecting the expression level of circDHTKD1 in preparing a kit for diagnosing and/or detecting airway epithelial inflammation.

The second technical scheme is as follows: application of a reagent for detecting the expression level of circDHTKD1 in preparation of a kit for diagnosing and/or detecting asthma.

Further, the circDHTKD1 regulates ERK pathway activation and inflammatory factor secretion through sponge adsorption miR-338-3p so as to realize diagnosis or detection of airway epithelial inflammation or asthma.

Further, the inflammatory factors are IL-6 and VEGF.

The third technical scheme is as follows: use of an inhibitor of circDHTKD1 in the manufacture of a medicament for modulating airway epithelial inflammation.

The technical scheme is as follows: application of an inhibitor of circDHTKD1 in preparation of medicine for treating asthma is provided.

Furthermore, the inhibitor takes circDHTKD1 as a target point, and achieves the purposes of regulating airway epithelial inflammation or treating asthma.

Compared with the prior art, the invention has the beneficial effects that:

the research result shows that the circDHTKD1 promotes the inflammatory reaction of the LPS-induced BEAS-2B cells, and the circDHTKD1 regulates an ERK pathway through miR-338-3p and influences the inflammatory reaction induced by the LPS. The invention proves that the circDHTKD1 is a key molecule for regulating and controlling airway epithelial inflammation, and the circDHTKD1 is expected to become a target for treating airway inflammation, so that a potential target for inhibiting the injury of the airway epithelial inflammation of a human is provided for clinic, and a theoretical basis is provided for the research of related medicine targets. The research result of the invention provides a new pathophysiological mechanism for the occurrence of the bronchial asthma, and can be used for preventing and treating the bronchial asthma.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 is a graph showing the results of analysis of the effect of LPS detection by CCK8 on the viability of BEAS-2B cells;

FIG. 2 shows the effect of LPS on the expression of BEAS-2B cell inflammatory factors (IL-6, IL-8, TNF-. alpha.and VEGF), where A is the relative expression level of mRNA and B is the protein expression level;

FIG. 3 is a graph showing the results of relative expression of circDHTKD by qRT-PCR;

FIG. 4 is a graph of Sanger sequencing results;

FIG. 5 is a graph showing the results of the nuclear and cytoplasmic separation assay for circDHTKD1 subcellular localization;

FIG. 6 shows the effect of qRT-PCR assays to knock down circDHTKD1 on the expression level of circDHTKD 1;

FIG. 7 shows the detection of inflammatory factor changes following the knockdown of circDHTKD1, where panel A is the change in IL-6 mRNA expression; panel B is a graph of detecting changes in mRNA expression of IL-8; panel C is a graph measuring changes in mRNA expression of TNF- α; graph D is the detection of mRNA expression changes in VEGF; e is a graph of detecting changes in the expression levels of inflammatory factor protein;

FIG. 8 is a schematic diagram of the target validation of circDHTKD/miR-338-3 p;

FIG. 9 is a graph of the results of a dual luciferase assay;

FIG. 10 is a panel of knockdown circDHTKD1 detecting changes in miR-338-3p expression levels;

FIG. 11 is a schematic diagram showing the results of detecting protein expression after miR-338-3p expression is changed;

FIG. 12 is a schematic representation of the detection of protein expression of ERK, P-ERK;

FIG. 13 is a graph showing the expression of IL-6 and VEGF proteins detected, wherein FIG. A is a graph showing the expression of VEGF proteins; FIG. B is a schematic representation of IL-6 protein expression.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but rather as a more detailed description of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in the present disclosure, it is understood that each intervening value, to the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated herein by reference to disclose and describe the methods and/or materials in connection with which the documents are cited. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

The Opti-MEM medium used in the examples was from Gibco; the dual-luciferase reporter gene detection kit adopts Beijing Quanzijin Biotechnology company, model FR 201; IL-6, IL-8, TNF- α and VEGF ELISA kits were purchased from Shanghai Bintian Biotechnology Ltd.

Example 1

Materials and methods

1. Test object

The human bronchial epithelial cell line (BEAS-2B) was purchased from the Shanghai cell Bank of the Chinese academy of sciences.

2. Experimental methods

2.1 main grouping:

(1) effect of LPS on cell viability: 0. mu.g/mL, 0.5. mu.g/mL, 1. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL.

(2) Effects of LPS on airway epithelial cells: control and LPS treated groups were divided.

(3) Effect of circDHTKD1 on gene expression: the test sample is divided into a control group, an LPS treatment group, a si-NC + LPS treatment group and a si-circ #1+ LPS group.

(4) The dual luciferase reporter gene experiments were grouped as follows: the method comprises the following steps of (1) circ-WT + miR-NC, (ii) circ-WT + miR-338-3p mimics, circ-MUT + miR-NC and circ-MUT + miR-338-3p mimics.

(5) Effect of miR-338-3p on Gene expression: the method comprises the following steps of firstly, preparing a reference group, secondly, processing LPS, thirdly, processing miR-NC + LPS and fourthly, miR-338-3p mimics + LPS.

(6) Functional rescue experiments were grouped as follows: the method comprises the following steps of firstly, preparing a reference group, secondly, preparing an LPS treatment group, thirdly, preparing an LPS + si-NC group, fourthly, preparing an LPS + si-circ #1 group, fifthl, preparing an LPS + si-circ #1+ miR inhibitor NC group and sixthly, preparing an LPS + si-circ #1+ miR-338-3p inhibitor group.

2.2 cell culture

BEAS-2B cells were cultured in RPMI-1640 medium (Thermo Fishier Scientific) supplemented with 10% bovine serum albumin (BSA; SigmaAldrich) and 100U/mL penicillin and 100. mu.g/mL streptomycin, at 5% CO₂The culture was carried out at 37 ℃ in a humid environment.

2.3 construction of LPS-induced bronchial epithelial cell inflammation model

And (3) taking BEAS-2B cells which are in a good growth state and are paved on a cell culture dish by more than 80%, digesting, centrifuging, discarding supernatant, adding 1mL of double-antibody-free RPMI-1640 complete culture medium for resuspension, and counting by using a cell counting plate. According to 5X 10 per hole⁵The individual cells were seeded in 6-well plates at 5X 10 per well⁴Inoculating the individual cells in a 96-well plate, shaking gently, mixing to make the cells uniformly distributed, placing the culture dish flat, placing at 37 deg.C and 5% CO₂Culturing in an incubator. When the cells in the culture plate were cultured to 80% confluency, the original medium was discarded, and different concentrations of LPS (0. mu.g/mL, 0.5. mu.g/mL, 1. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL) were added to the culture medium for further culture for 24 hours.

2.4 CCK8 determination of cell viability

Adjusting the concentration of cell suspension to 510⁵one/mL, adding 100 μ L of each well into a 96-well cell culture plate, setting a blank control group (only adding 100 μ L of culture medium), and setting three multiple wells for the same sample; adding 100 mu L PBS into peripheral holes at the periphery to reduce errors, culturing for 24h in a 5% incubator at 37 ℃, removing the original culture medium, taking 100 mu L of newly prepared culture medium (0 mu g/mL, 0.5 mu g/mL, 1 mu g/mL, 5 mu g/mL, 10 mu g/mL and 20 mu g/mL) containing LPS with different concentrations for each group, adding the culture medium containing the PBS with the same volume as the control group, continuing culturing for 24h, adding 10 mu L CCK-8 reagent into a super clean bench in a dark place, shaking gently and mixing the solution, placing the solution into the incubator for continuing 1-2 h, opening an enzyme labeling instrument to preheat the solution to 37 ℃, and measuring the absorbance at the wavelength of 450 nm.

2.5 cell transfection

One day before transfection, cells were treated according to the above method, and 5X 10 cells were subjected to density adjustment using antibiotic-free RPMI-1640 complete medium⁵And inoculating the cells into a 6-hole cell culture plate, and culturing overnight to ensure that the cell confluency reaches 50-70% the next day. The original medium was aspirated off and the cells were gently rinsed twice with PBS. Add 1.5mL of Opti-MEM medium per well. Two 1.5mL enzyme-free EP tubes, labeled A and B, were used, and 125. mu.L of Opti-MEM medium was added to each of the A, B tubes, and the wells were prepared in bulk. Add 5. mu.L Lipofectamine to EP labeled A^TM3000, blowing and beating by a pipettor and uniformly mixing; and adding 5 mu L of si-RNA or miR-imic/miR-inhibitor into the EP tube marked with the B, and blowing and beating by a pipette to be uniformly mixed. Mixing the solution in the tube A and the tube B, blowing and stirring uniformly, and standing at room temperature for 15 min. Adding 250 μ L of the mixed solution into corresponding cell wells, shaking, mixing, and standing at 37 deg.C with 5% CO₂After culturing for 8h in the incubator, the transfection medium is discarded, and the complete medium without antibiotic is replaced by RPMI-1640 to continue culturing. Followed by LPS treatment. All RNA sequences used in the transfection experiments are shown in table 1.

TABLE 1 RNA sequences for cell transfection experiments

2.6 RT-PCR and quantitative RT-PCR

Total cellular RNA was extracted with RNAasso PLUS according to PrimeScript^TMThe RTReagent kit instructions formulate a reverse transcription reaction system (as shown in table 2), with the reverse transcription program set as follows: 15min at 37 ℃ (reverse transcription); 5sec at 85 ℃ (inactivation reaction of reverse transcriptase); infinity at 4 ℃. According to TB

Premix Ex Taq^TMThe kit instruction prepares a reaction system (shown in table 3), and the real-time fluorescent quantitative PCR reaction program is as follows: pre-denaturation at 95 ℃ for 30 s; PCR reaction at 95 deg.C for 5s for 40 cycles; annealing the primer at 60 ℃ for 30 s; primer extension 60 ℃ for 60 s. After the reaction is finished, CT value is obtained, and 2 is adopted^-ΔΔctThe relative expression of mRNA was analyzed and statistically analyzed (see FIG. 1 for results).

Primer sequences used in the experiments (as shown in Table 4)

TABLE 220 μ L reverse transcription reaction System

Name of reagent	Amount of addition
		5×PrimeScript^TM Buffer	4μL
PrimeScript^TM RT Enzyme Mix	1μL
		Oligo dT Primer(50μM)	1μL
Random
	6 mers(100μM)	1μL
Total RNA			1μL
	RNase-free Water	12μL
Total			20μL

Table 320 mu L real-time fluorescent quantitative PCR reaction system

TABLE 4 primers used in RT-qPCR experiments

2.7 Dual luciferase reporter Gene detection

A wild-type circ-WT fragment containing the full length of circDHTKD1 and having a miR-338-3p binding site, and a mutant circ-MUT fragment with a mutated binding site were synthesized and inserted into the psiCHECK2 vector. Wild type and mutant dual-luciferase reporter plasmids, miR-338-3p-mimic or mimic NC and Lipofectamine^TM3000 cells were co-transfected with the formulated complex, as described above. After 48 hours of incubation, cells were collected and lysed and Relative Light Units (RLU) of firefly luciferase and renilla luciferase were determined using the dual-luciferase assay kit.

One day before transfection, cells were treated according to the above method, and 5X 10 cells were subjected to density adjustment using antibiotic-free RPMI-1640 complete medium⁵And (3) inoculating each cell in a 6-hole cell culture plate, and culturing overnight to ensure that the cell confluency reaches 50-70% on the next day. The original medium was aspirated off, and gently moistened with PBSCells were washed twice. 1.5mL of Opti-MEM medium was added to each well. Two 1.5mL enzyme-free EP tubes, labeled A and B, were used, and 125. mu.L of Opti-MEM medium was added to each of the A, B tubes, and the wells were prepared in bulk. Add 5. mu.L Lipofectamine to EP labeled A^TM3000, blowing and beating by a pipettor and uniformly mixing; and adding 5 mu L of si-RNA or miR-imic/miR-inhibitor into the EP tube marked with the B, and blowing and beating by a pipette to be uniformly mixed. Mixing the solution in the tube A and the tube B, blowing and stirring uniformly, and standing for 15min at room temperature. Adding 250 μ L of the mixed solution into corresponding cell wells, shaking, mixing, and standing at 37 deg.C with 5% CO₂After culturing for 8h in the incubator, the transfection medium is discarded, and the complete medium without antibiotic is replaced by RPMI-1640 to continue culturing. Followed by LPS treatment. All RNA sequences used in the transfection experiments are shown in table 1.

2.8 Western blot

Total protein was extracted from cultured BEAS-2B cells using RIPA lysate. Protein concentration was determined by BCA protein assay kit produced in petunia. Subsequently, the proteins were mixed with 5 Xprotein loading buffer and boiled at 95 ℃ for 5min, separated by SDS-PAGE electrophoresis and transferred to PVDF membrane. Blocking with 5% bovine serum albumin solution at room temperature for 2h, then incubating the PVDF membrane with protein and the diluted primary antibody at 4 ℃ overnight, washing the membrane with TBST for 3 times, incubating the secondary antibody and the PVDF membrane at room temperature for 2h, washing the membrane with TBST for 3 times, developing with ECL luminescence solution, taking a picture with a gel imaging system, and storing. The gene GAPDH was normalized as an internal control.

2.9 extraction of RNA by nucleoplasm separation

According to PARIS^TMThe kit is used for separating and extracting total RNA of cytoplasm and nucleus of BEAS-2B cells. The subcellular localization of circRNA was performed by qRT-PCR experiments. GAPDH and NEAT1 were used as cytoplasmic and nuclear controls, respectively.

2.10 enzyme-linked immunosorbent assay (ELISA)

BEAS-2B cells at 2X 10 per well⁵The density of individual cells was seeded in 12-well plates. The cell culture dish supernatant was collected from a sterile 1.5mL EP tube, centrifuged at 2000rpm for 20 minutes, and the supernatant was collected. According to the kit specification, the inflammatory cytokines IL-6 and IL-8. Levels of TNF-alpha and VEGF. OD measurement at 450nm wavelength with full-function microplate reader₄₅₀The value is obtained.

2.11 nucleic acid electrophoresis and Sanger sequencing

And (3) carrying out electrophoretic separation on the PCR experimental product in 2% agarose gel, observing the product by an ultraviolet imaging system, cutting the gel, and sequencing the gel by Shanghai biological engineering Co.

2.12 statistical analysis method

The present study analyzed quantitative data using SPSS 20.0 software, expressed as mean ± standard deviation. Differences between the two or more groups were assessed by Student T-test or ANOVA (analysis of variance) methods. Significant differences were considered to exist at P values < 0.05.

Model of inflammation induced by LPS (LPS) in BEAS-2B cells

3.1 dilution of LPS with RPMI-1640 medium containing 10% FBS without diabody to different concentrations (0. mu.g/mL, 0.5. mu.g/mL, 1. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, 20. mu.g/mL), BEAS-2B cells were cultured for 12 h. The CCK-8 method measures the effect of different concentrations of HDM on cell proliferation. As shown in FIG. 1, LPS at a concentration of 5. mu.g/mL or more significantly inhibited the activity of BEAS-2B cells, as compared to the control group.

3.2 the experimental group stimulated the cells with 5. mu.g/mL LPS for 24h, the control group was given an equal volume of PBS, RNA was extracted and then mRNA expression levels of IL-6, IL-8, TNF-. alpha.and VEGF were determined by real-time fluorescent quantitative PCR. As shown in FIG. 2, the relative mRNA expression levels of IL-6, IL-8, TNF-. alpha.and VEGF and the supernatant protein content were significantly increased in the LPS group compared to the control group.

The circDHTKD1 promotes LPS-induced inflammation of BEAS-2B cells.

4.1 qRT-PCR results As shown in FIG. 3, LPS treatment resulted in increased expression of circDHTKD1 compared to the negative control group.

4.2 circDHTKD1 localizes the cytoplasm, which was confirmed by Sanger sequencing to be a circular RNA molecule formed by reverse splicing. The Sanger sequencing results show the sequence containing the reverse splice site, indicating that the product is a circular RNA (see figure 4 for details). In addition, subcellular localization of circDHTKD1 in BEAS-2B cells was examined by nuclear matter isolation experiments, and it was found that circDHTKD1 was significantly enriched in the cytoplasm of BEAS-2B cells (see FIG. 5 for details)

4.3 role of circDHTKD1 in LPS-induced inflammatory injury of BEAS-2B cells: design 2 Gene expression interference was performed on siRNAs targeting circDHTKD1(si-circ #1 and si-circ # 2).

Design 2 siRNA targeting circDHTKD1(si-circ #1 and si-circ #2) was subjected to gene expression interference, and qRT-PCR was performed 48h after transfection to detect the expression of circDHTKD 1. The results are shown in detail in FIG. 6, where the expression level of circDHTKD1 was significantly reduced after transient transfection of these 2 pairs of siRNAs for 48h compared to si-NC, where the downregulation of the circDHTKD1 level by transfection of si-circ #1 was more pronounced. Si-circ #1 was chosen for subsequent experiments.

4.4 interference with circDHTKD1 expression and LPS stimulation for 24 hours, significant reductions were observed in both IL-6 and VEGF mRNA expression levels in BEAS-2B cells and protein expression levels in cell culture supernatants (see FIG. 7 for details).

circDHTKD1 as a sponge for miR-338-3p

5.1 bioinformatics predicted that circDHTKD1 has a miR-338-3p binding site (see FIG. 8 for details), and dual-luciferase reporter gene experiments confirmed that circDHTKD1 and miR-338-3p have a binding relationship. After miR-338-3p micic and circ-WT/circ-MUT plasmid transfect cells, miR-338-3p is found to obviously reduce the activity of circ-WT luciferase, and has no inhibition effect on the activity of circ-MUT luciferase. Suggesting that circDHTKD1 can be targeted to bind to miR-338-3p (see FIG. 9 for details).

5.2 after transfection of siRNA in BEAS-2B cells to interfere with the expression of circRNA, the relative expression level of miR-338-3p is up-regulated (see FIG. 10 for details).

circDHTKD1 regulating ERK pathway through miR-338-3p and influencing inflammatory reaction induced by LPS

6.1 miR-338-3p targets the ERK signaling pathway: after miR-338-3p mimic is transfected, the expression level of miR-338-3p in cells is obviously increased. When cells transfected with miR-338-3p mimic are stimulated with LPS for 24 hours again, the protein levels of IL-6 and VEGF are reduced remarkably, and the phosphorylation level of ERK protein is also reduced (see figure 11 for details).

6.2 circDHTKD1/miR-338-3p axis regulates ERK protein phosphorylation level and IL-6 and VEGF synthesis secretion: after the circDHTKD1 siRNA and miR-338-3p inhibitor are co-transfected into BEAS-2B cells respectively, compared with an LPS + si-NC group, the ERK phosphorylation level of the LPS + si-circ #1 group is reduced; the p-ERK protein level was elevated in the LPS + si-circuit #1+ miR-338-3p inhibitor group compared to the LPS + si-circuit #1+ miR inhibitor NC group. Knockdown circDHTKD1 inhibited LPS-induced activation of the ERK signaling pathway, and miR-338-3p inhibitor transfected to reverse the decreased level of ERK protein phosphorylation caused by circDHTKD1 knockdown (see FIG. 12 for details). By detecting the expression of IL-6 and VEGF in cell supernatants through ELISA experiments, miR-338-3p inhibitor transfected can reverse the reduction of IL-6 and VEGF protein expression caused by the knockdown of circDHTKD1 (see FIG. 13 for details).

The present study shows that circDHTKD1 regulates ERK pathway activation and inflammatory factor secretion by sponge adsorption of miR-338-3p in LPS-treated BEAS-2B cells.

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Claims

1. Application of an agent for detecting the expression level of circDHTKD1 in preparing a kit for diagnosing and/or detecting airway epithelial inflammation.

2. Application of an agent for detecting the expression level of circDHTKD1 in preparing a kit for diagnosing and/or detecting asthma.

3. The use of any of claims 1-2, wherein the circDHTKD1 is used to modulate ERK pathway activation and inflammatory factor secretion by sponge adsorption of miR-338-3p to diagnose or detect airway epithelial inflammation or asthma.

4. The use according to claim 3, wherein the inflammatory factors are IL-6 and VEGF.

5. Use of an inhibitor of circDHTKD1 in the manufacture of a medicament for modulating airway epithelial inflammation.

6. Application of an inhibitor of circDHTKD1 in preparation of medicines for treating asthma is provided.

7. The use according to any one of claims 5 to 6, wherein the inhibitor targets circDHTKD1 for the purpose of modulating airway epithelial inflammation or treating asthma.