Application of ADRB2 agonist in preparation of medicine for preventing and / or treating sepsis

Abstract

The invention relates to the field of medicines, in particular to application of an ADRB2 agonist in preparation of a medicine for preventing and / or treating sepsis. Cell experiments and animal experiments find that expression of IL-10 in a sepsis model mouse body can be hindered and the death rate can be increased by knocking out Adrb2 in B cells, and the death rate of the sepsis model mouse can be reduced by administering an ADRB2 activating agent. The ADRB2 agonist can be used for preventing and / or treating sepsis, and a new thought is provided for clinical medication of sepsis.

Description

Technical Field

[0001] The present invention relates to the field of medicines, and in particular, to the use of an ADRB2 agonist in preparing a medicine for preventing and / or treating sepsis. Background Art

[0002] In cases where organs (such as the lungs and digestive tract) are severely infected or pathogens (such as LPS) enter the blood in large quantities, the immune system is overactivated and sepsis occurs. In these pathological processes, the immune response releases excessive amounts of inflammatory factors such as TNFa, IL-6, IFN-g, or inflammatory substances such as reactive oxygen species, causing multiple organ damage and disseminated intravascular coagulation, and even death. Currently, there is a lack of treatment options for sepsis, and it is almost impossible to predict or prevent the occurrence of this fatal disease clinically.

[0003] ADRB2 receptors are widely expressed in immune cells, and studies have suggested that they may be associated with a variety of human diseases. Currently, β-receptor agonists have been used as successful clinical applications to improve airway obstruction caused by asthma, chronic obstructive pulmonary disease, and lung dysfunction associated with pulmonary fibrosis. However, despite the high attention paid to the potential therapeutic value of ADRB2 in a variety of diseases, there are currently no reports on its application in the treatment of sepsis. This suggests that the specific mechanism of action of ADRB2 in sepsis and its therapeutic potential still need further study, providing a possible direction for the development of new therapeutic strategies. Summary of the invention

[0004] In order to fill the gap in the prior art, the present invention provides an application of an ADRB2 agonist in the preparation of a medicament for preventing and / or treating sepsis.

[0005] In one embodiment, the use comprises enhancing IL-10 expression in regulatory B cells by using an ADRB2 agonist.

[0006] In one embodiment, the use includes reducing a patient's disease susceptibility to sepsis by using an ADRB2 agonist.

[0007] In one embodiment, the use includes directly promoting the anti-inflammatory function of IL-10 positive regulatory B cells via the ADRB2 receptor.

[0008] In a preferred embodiment, the ADRB2 agonist is formoterol. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0010] Figure 1 : After LPS-induced sepsis in Cd19-Cre; Adrb2 fl / fl and littermate Adrb2 fl / fl control mice, qPCR was used to detect the IL-10 mRNA levels in the spleen, liver, and inguinal lymph nodes of mice 1 hour after LPS treatment;

[0011] Figure 2 : After LPS-induced sepsis in Cd19-Cre; Adrb2 fl / fl and littermate Adrb2 fl / fl control mice, ELISA was used to detect the IL-10 protein level in mouse plasma 1 hour after LPS treatment;

[0012] Figure 3 :After LPS-induced sepsis in Cd19-Cre; Adrb2 fl / fl and littermate Adrb2 fl / fl control mice, the survival rate of the mice was measured daily;

[0013] Figure 4 : CD45+CD19+CD24hi CD27+B cells were sorted from human spleen and divided into three groups: saline-treated group (control group), LPS-treated group (B cells under simulated sepsis conditions) and LPS+formoterol-treated group (B cells under simulated sepsis and treated with formoterol). The expression levels of IL-10 mRNA and protein were detected by qPCR and ELISA.

[0014] Figure 5 : After splenic sympathectomy, C57BL / 6 wild-type mice were treated with LPS to induce sepsis. Then, the mice were treated with normal saline (control group) or formoterol (experimental group). qPCR was used to detect the IL-10 mRNA level in the spleen of mice 1 hour after LPS treatment. ELISA was used to detect the IL-10 protein level in the plasma of mice 1 hour after LPS treatment.

[0015] Figure 6 : After splenic sympathectomy, C57BL / 6 wild-type mice were treated with LPS to induce sepsis. The mice were then treated with normal saline (control group) or formoterol (experimental group), and the survival rate of the mice was measured daily. DETAILED DESCRIPTION

[0016] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0017] Example 1 Establishment of mouse sepsis model

[0018] Cd19-Cre; Adrb2 was obtained by crossing B cell-specific tool mouse Cd19-Cre mice (Saiye Biotechnology, #C001204) with Adrb2 fl / fl mice (Saiye Biotechnology, #S-CKO-01084). fl / fl Mouse strains that achieve specific deletion of Adrb2 in B cells.

[0019] For the LPS-induced sepsis model, LPS (dissolved in sterile PBS at a concentration of 1 mg / mL) was intravenously injected at 10 mg / kg body weight into mice of the specified conditions (including wild-type mice (C57BL / 6J) in the control group and CD19-Cre; Adrb2 fl / fl B cell conditional knockout mice).

[0020] For the cecal ligation and puncture (CLP)-induced sepsis model, mice of the indicated conditions were anesthetized with 3% isoflurane, and the abdominal skin was shaved and treated with iodine and alcohol. A longitudinal incision was made along the midline of the skin and peritoneum to expose the cecum. The cecum was ligated with 5 / 0 silk thread approximately 1 cm from the distal end. The distal end of the ligated cecum was punctured twice with a 22-gauge needle. The sham operation included all the steps except that the cecum was not ligated or punctured. The incisions on the peritoneum and skin were then sutured.

[0021] After the animal model was established, the levels of cytokines (such as IL-10, TNF-α, IFN-β) in the spleen and plasma were detected by qPCR and ELISA. Significant changes in IL-10 levels are an important indicator of the success of the endotoxemia model. In addition, changes in the spleen immune microenvironment were observed by immunolabeling techniques (such as TH marking sympathetic nerves and CD169 marking marginal zone macrophages). More importantly, the survival rate of the experimental animals was recorded to evaluate the severity of sepsis and the success of the model. A decrease in survival rate indicates the establishment of sepsis.

[0022] Example 2 Specific knockout of the Adrb2 receptor on B cells in sepsis model mice leads to increased mortality in mice

[0023] Compared with the control mice (C57BL / 6J) treated with LPS, Cd19-Cre; Adrb2 fl / fl The expression of IL-10 in the spleen of mice was significantly inhibited after LPS treatment, while the expression level of IL-10 in the liver or inguinal lymph nodes did not seem to be affected ( Figure 1 This defective splenic IL-10 response resulted in reduced plasma IL-10 levels ( Figure 2 ), resulting in decreased survival rate of Cd19-Cre; Adrb2 fl / fl mice ( Figure 3 ).

[0024] The above results indicate that specific knockout of the Adrb2 receptor on B cells in sepsis model mice leads to increased mortality in mice.

[0025] Example 3 Adrb2 agonist formoterol can promote the expression of IL-10 in human B cells

[0026] CD45 Isolation from Human Spleens by Flow Cytometry + CD19 + CD24 hi CD27 + B cells were divided into three groups: saline treatment group (control group), LPS treatment group (B cells under simulated sepsis conditions) and LPS + formoterol treatment group (B cells under simulated sepsis and treated with formoterol). The expression of IL-10 mRNA and protein was detected by qPCR and ELISA. It was found that the ADRB2 agonist formoterol effectively enhanced the LPS-induced human CD24hi CD27+ B cell IL-10 mRNA expression and protein production ( Figure 4 ). This indicates that formoterol directly promotes the anti-inflammatory function of IL-10 positive regulatory B cells through the ADRB2 receptor.

[0027] Example 4 Adrb2 agonist formoterol can inhibit the mortality of septic mice

[0028] After LPS-induced sepsis in mice, formoterol (Selleckchem, dissolved in 0.5 mg / ml sterile PBS) was immediately injected intravenously at a dose of 5 mg / kg body weight. Compared with the control group (LPS-induced C57BL / 6J wild-type mice), this Adrb2 agonist promoted the production and secretion of IL-10 in mice ( Figure 5 ), significantly inhibited the mortality of septic mice and completely restored the disease susceptibility of these mice ( Figure 6 ).

[0029] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Use of an ADRB2 agonist in the preparation of a medicament for preventing and / or treating sepsis.

2. The use according to claim 1, characterized in that The application includes enhancing IL-10 expression in regulatory B cells by using an ADRB2 agonist.

3. The use according to claim 1, characterized in that The use includes reducing a patient's disease susceptibility in sepsis by using an ADRB2 agonist.

4. The use according to claim 1, characterized in that The application includes directly promoting the anti-inflammatory function of IL-10 positive regulatory B cells through the ADRB2 receptor.

5. The use according to claim 1, wherein the ADRB2 agonist is formoterol.

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