Trace CSE-modified polypeptide gold nanoparticles and application thereof in preparation of drugs for treating acute lung inflammatory diseases

Abstract

The invention relates to an application of trace CSE modified polypeptide gold nanoparticles in preparation of drugs for treating acute lung inflammatory diseases. The polypeptide-modified gold nanoparticle is a polypeptide-modified gold nanoparticle as shown in a sequence SEQ ID NO: 1. The invention proves that the gold nanoparticle compound CSE-P12 of polypeptide (CLPFFD, SEQ ID NO: 1) modified by trace CSE (0.1%-1%) can degrade STING by inducing autophagy of macrophages for the first time, so that a cGAS-STING signal channel is inhibited to play a role in anti-inflammatory activity, and the gold nanoparticle compound CSE-P12 plays a role in efficiently protecting an acute lung injury model induced by DNA (deoxyribonucleic acid) virus (adenovirus HAdV4), RNA (ribonucleic acid) virus (influenza virus PR8) and bacterial sepsis. Therefore, the compound can be used for preparing medicines for treating acute lung inflammation and injury diseases caused by infection. The medicine has the advantages of being remarkable in curative effect, capable of acting on macrophages in the lung and other tissues in a targeted mode, convenient to track in vivo and the like, and a new treatment means is provided for acute lung inflammation injury caused by infection.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and more specifically, to the use of polypeptide gold nanoparticles modified with trace CSE (0.1%-1%) in the preparation of drugs for treating acute pulmonary inflammatory diseases induced by adenovirus, influenza virus, and sepsis. Background Art

[0002] Acute lung injury (ALI) and its more severe form, acute respiratory distress syndrome (ARDS), are acute progressive hypoxic respiratory failures caused by various pathogenic factors inside and outside the lungs other than cardiogenic factors. Bacterial and viral pneumonia, as well as non-pulmonary infections (such as sepsis and trauma), are common causes of acute pulmonary inflammation. The main pathogenic feature of such diseases is the uncontrolled inflammatory response in the lungs, such as cytokine storms and inflammatory cell infiltration, leading to increased pulmonary capillary permeability and diffuse alveolar damage. Bacterial and viral infections, as well as particulate poisons, can induce host cell stress, cell death, and tissue damage, resulting in the release of self-DNA in the intracellular or extracellular environment. Nuclear DNA or mtDNA can abnormally activate the cGAS-STING signaling pathway in macrophages, leading to a strong type I interferon (IFN-I) response and the release of pro-inflammatory factors, thereby exacerbating the pulmonary inflammation induced by viral and bacterial infections. Currently, the main drugs used to treat acute lung injury include those that inhibit the inflammatory response (corticosteroids, N-acetylcysteine, macrolides, statins, and sivelestat), reduce pulmonary interstitial or alveolar edema (pulmonary surfactant and β2-receptor agonists), promote pulmonary vasodilation (nitric oxide and prostacyclin), act on the coagulation pathway (aspirin and platelet-activating factor receptor antagonists), and promote the repair of alveolar epithelial cells and vascular endothelial cells (keratinocyte growth factor). However, these drugs have not reduced the mortality rate of ALI in clinical trials. Summary of the Invention

[0003] The object of the present invention is to provide the use of polypeptide gold nanoparticles modified with trace CSE (0.1%-1%) in view of the deficiencies in the prior art.

[0004] A polypeptide gold nanoparticle modified with trace cigarette smoke extract CSE, wherein the trace CSE modifies its sequence SEQ ID NO: 1, and the polypeptide gold nanoparticle is designated as CSE-P12. Trace CSE refers to 0.1%-1% cigarette smoke extract.

[0005] The use of a polypeptide gold nanoparticle modified with trace CSE in inhibiting the cGAS-STING signaling pathway in vitro.

[0006] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in inducing autophagy in macrophages and degrading STING through autophagy

[0007] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by DNA virus, adenovirus HAdV4

[0008] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by RNA virus, influenza virus PR8, wherein the polypeptide gold nanoparticles are gold nanoparticles modified by a polypeptide with the sequence shown in SEQ ID NO:1

[0009] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in a drug for treating acute lung injury induced by sepsis bacterial infection

[0010] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by DNA virus, wherein the DNA virus is adenovirus

[0011] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by DNA virus, wherein the adenovirus is HAdV4

[0012] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by RNA virus, wherein the RNA virus is influenza virus

[0013] Application of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute lung inflammation induced by RNA virus, wherein the influenza virus is PR8

[0014] The DNA virus proposed in this application is human adenovirus (HAdV). The HAdV related to human respiratory tract infection mainly includes subgroup B (HAdV-3, 7, 11, 14, 16, 21, 50, 55 types), subgroup C (HAdV-1, 2, 5, 6, 57 types) and subgroup E (HAdV-4 type), etc. The DNA virus proposed in this application also includes canine adenovirus (CAV), porcine adenovirus (PAV), ovine adenovirus (OAV) and bovine adenovirus (BAV), etc

[0015] The RNA virus proposed in this application is influenza virus, such as influenza A virus and influenza B virus, etc

[0016] The trace amount of CSE (0.1%-1%) modifies the polypeptide gold nanoparticles with the sequence shown in SEQ ID NO:1, and the code name is CSE-P12

[0017] Polypeptide P12: CLPFFD (SEQ ID NO:1) is a short-chain polypeptide composed of five amino acids. The following is a detailed explanation of this polypeptide:

[0018] Amino acid sequence: CLPFFD consists of the following amino acids:

[0019] C: Cysteine (C)

[0020] L: Leucine (L)

[0021] P: Proline (P)

[0022] F: Phenylalanine (F)

[0023] SEQ ID NO:1: This represents the unique identifier of this sequence in the bioinformatics database, used to identify and reference a specific sequence.

[0024] The preparation method of polypeptide gold nanoparticles modified with trace CSE (0.1%-1%) is as follows:

[0025] (1) Synthesis of gold nanoparticles:

[0026] The synthesis of gold nanoparticles uses the method of reducing chloroauric acid with sodium citrate. The specific operation method is as follows:

[0027] Add 161.2 μL of 420 mg / mL chloroauric acid to a conical flask containing 200 mL of 18.2 MΩ ultrapure water. After heating to boiling on a magnetic stirrer, quickly add 2 mL of 10% sodium citrate, continue heating and stirring for 15 minutes, and then stir and cool to room temperature.

[0028] Polypeptide P12: CLPFFD (SEQ ID NO:1) powder with a purity >95% is dissolved in ultrapure water to prepare a polypeptide solution (1 mM);

[0029] Mix the polypeptide and gold nanoparticle solution in a volume ratio of 1:10 and let it stand at room temperature for 24 h to allow the polypeptide and gold nanoparticles to fully bind;

[0030] Filter the polypeptide gold nanoparticle suspension using a 0.22 μm syringe filter head, then perform centrifugation at 15000 rpm for 30 min, and then wash three times with PBS and concentrate to 100 nM (nanoparticle concentration) for use;

[0031] (2) Preparation of cigarette smoke extract CSE:

[0032] Obtain CSE using commercially available cigarettes containing 10 mg of tar and 0.8 mg of nicotine,

[0033] The specific operation method is as follows:

[0034] First, seal and connect two 10 mL gas sampling bottles through a rubber tube. Fix a cigarette with the filter tip cut off to one side of the first gas sampling bottle using the rubber tube. Add 10 mL of RPMI 1640 medium to this gas sampling bottle. Connect a 50 mL syringe to one side of the second gas sampling bottle through the rubber tube. After the sealed connection is completed, light the cigarette and quickly draw air using the 50 mL syringe. The cigarette smoke dissolves in the 10 mL RPMI 1640 medium to form 100% CSE of cigarette extract. Then filter and dilute it, and fully mix and incubate it with the polypeptide gold nanocomposite (P12) for 1 h, finally obtaining CSE-P12 containing 0.1% - 1% CSE.

[0035] (3) Modify the polypeptide gold nanoparticles with trace CSE

[0036] It can be achieved by physical adsorption, covalent binding, surface modification, etc.

[0037] The beneficial effects of the present invention are as follows:

[0038] The present invention for the first time proves that the polypeptide (CLPFFD, SEQ ID NO:1) gold nanoparticle complex CSE-P12 modified by trace CSE (0.1% - 1%) can inhibit the cGAS-STING signaling pathway and exert anti-inflammatory activity by inducing autophagy degradation of STING in macrophages, and can play a highly protective role in acute lung injury models induced by DNA virus (adenovirus HAdV4), RNA virus (influenza virus PR8), and bacterial sepsis. Therefore, it can be used to prepare drugs for treating acute pulmonary inflammation and injury diseases caused by infection. This drug has the advantages of significant curative effect, being able to target macrophages in the lungs and other tissues, and being convenient for in vivo tracking, providing a new treatment means for acute pulmonary inflammation and injury caused by infection.

[0039] Pathogenic infections such as bacteria and viruses, sepsis, or other inducing conditions such as immunotherapy can cause excessive stress in the body's immune cells, resulting in the production of excessive cytokines, especially type I interferons, which exacerbate the inflammatory response and induce acute pulmonary inflammatory diseases, including pneumonia caused by novel coronavirus infection, adenovirus pneumonia, influenza virus pneumonia, and acute lung injury caused by sepsis bacterial infection. These diseases have high morbidity and mortality, and uncontrolled inflammatory response is their main feature, and there are no effective therapeutic drugs yet. The present invention for the first time confirms that CSE-P12 can inhibit the cGAS-STING signaling pathway by inducing autophagy to degrade STING, and can reduce DNA virus (adenovirus HAdV4), RNA virus (influenza virus PR8) induced pulmonary inflammatory injury and sepsis-related acute lung injury. Therefore, it can become an effective drug for treating acute pulmonary inflammatory diseases caused by abnormal activation of cGAS-STING. This drug has the advantages of significant efficacy, being able to target specific cells, and being convenient for in vivo tracking, etc., providing a new treatment means for ALI.

[0040] The present invention provides a polypeptide-based nanoagent, which is a novel pharmaceutical preparation that has been studied more in recent years. It has a series of advantages such as being able to target specific cells, increasing the selectivity of drug delivery, improving the treatment efficiency, and reducing side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figures 1(a) - 1(c) show that CSE-P12 inhibits the cGAS-STING signaling pathway in THP-1-derived macrophages.

[0042] Figure 1(a) shows the activation of IRF after CSE-P12 stimulates macrophages derived from THP-1 reporter cells with cGAMP for 24 h (n = 3);

[0043] Figure 1(b) shows the activation of NF-κB / AP-1 after CSE-P12 stimulates macrophages derived from THP-1 reporter cells with cGAMP for 24 h (n = 3);

[0044] Figure 1(c) shows the gene expression of MX1 after CSE-P12 stimulates THP-1-derived macrophages with cGAMP for 24 h (n = 4);

[0045] Figures 2(a) - 2(b) show that CSE-P12 induces autophagy in THP-1-derived macrophages and degrades STING through autophagy.

[0046] Figure 2(a) shows the expression of autophagy-related proteins p62 and LC3II after CSE-P12 stimulates THP-1-derived macrophages with cGAMP for 4 h;

[0047] Figure 2(b) Expression of STING protein after cGAMP-stimulated THP-1-derived macrophages were treated with CSE-P12 and Rapamycin for 4 h;

[0048] Figures 3(a)-3(b) show that CSE-P12 can effectively inhibit mouse pulmonary inflammation induced by adenovirus (HAdV4) infection;

[0049] Figure 3(a) Expression of type I interferon gene Ifnb1 in lung tissues of mice in the mouse pulmonary inflammation model induced by adenovirus (HAdV4) infection in the CSE-P12 treatment group;

[0050] Figure 3(b) Protective effect of CSE-P12 on pathological damage of lung tissues of mice infected with HAdV4 in the mouse pulmonary inflammation model induced by adenovirus (HAdV4) infection;

[0051] Figures 4(a)-4(b) show that CSE-P12 can effectively inhibit mouse pulmonary inflammation induced by influenza virus (PR8) infection;

[0052] Figure 4(a) Effect of CSE-P12 on the survival rate of mice infected with PR8 in the mouse pulmonary inflammation model induced by influenza virus (PR8) infection;

[0053] Figure 4(b) Protective effect of CSE-P12 on pathological damage of lung tissues of mice infected with PR8 in the mouse pulmonary inflammation model induced by influenza virus (PR8) infection;

[0054] Figures 5(a)-5(c) show that CSE-P12 improves the survival rate and pulmonary inflammation of mice in the CLP-induced acute lung injury mouse model;

[0055] Figure 5(a) Effect of CSE-P12 on the survival rate of mice in the CLP-induced mouse acute lung injury model;

[0056] Figure 5(b) Effect of CSE-P12 on the bacterial load in the spleen tissues of mice in the CLP-induced mouse acute lung injury model;

[0057] Figure 5(c) Effect of CSE-P12 on pathological damage of lung tissues of mice in the CLP-induced mouse acute lung injury model (n = 5); Detailed implementation manners

[0058] The following provides a detailed description of the specific implementation manners provided by the present invention in conjunction with the accompanying drawings.

[0059] Example 1

[0060] Preparation of CSE-P12

[0061] 1. Experimental method

[0062] The synthesis of gold nanoparticles was carried out by the method of reducing chloroauric acid with sodium citrate. The specific operation method is as follows: Add 161.2 μL of chloroauric acid (420 mg / mL) to a conical flask containing 200 mL of ultrapure water (18.2 MΩ). After heating to boiling on a magnetic stirrer, quickly add 2 mL of sodium citrate (10%). Continue heating and stirring for 15 minutes, and then stir and cool to room temperature. Polypeptide P12: CLPFFD (SEQ ID NO: 1), with a purity > 95% powder, was dissolved in ultrapure water to prepare a polypeptide solution (1 mM). The polypeptide and the gold nanoparticle solution were mixed at a volume ratio of 1:10 and allowed to stand at room temperature for 24 h to allow the polypeptide and the gold nanoparticles to fully bind. The polypeptide-gold nanoparticle suspension was filtered through a 0.22 μm syringe filter head, then centrifuged (15000 rpm, 30 min), and then washed three times with PBS and concentrated to 100 nM (nanoparticle concentration) for use.

[0063] To prepare cigarette smoke extract (CSE), commercially available cigarettes (containing 10 mg of tar and 0.8 mg of nicotine) were used to obtain CSE. The specific operation method is as follows: First, two 10 mL gas sampling bottles were hermetically connected through a rubber tube, and a cigarette with the filter tip cut off was fixed on one side of the first gas sampling bottle using a rubber tube. 10 mL of RPMI 1640 medium was added to this gas sampling bottle, and a 50 mL syringe was connected to the other side of the second gas sampling bottle through a rubber tube. After the hermetic connection was completed, the cigarette was lit, and the 50 mL syringe was used to quickly draw air. The cigarette smoke was dissolved in 10 mL of RPMI 1640 medium to form 100% CSE of cigarette smoke extract. Then, it was filtered, diluted, and fully mixed and incubated with the polypeptide-gold nanoparticle complex (P12) for 1 h to finally obtain CSE-P12 containing 0.1% - 1% CSE.

[0064] Example 2

[0065] CSE-P12 can inhibit the activation of the cGAS-STING signaling pathway

[0066] 1. Experimental method

[0067] To detect the activation of IRF and NF-κB / AP-1 in the cGAS-STING downstream signaling pathway, reporter cells THP-1-ISG or THP-1-XBlue cells were seeded in 96-well culture plates and differentiated into macrophages with 50 ng / mL phorbol 12-myristate 13-acetate (PMA); after 24 h, they were washed twice with PBS and left standing for 48 h before the experiment. Cells were treated with the STING agonist cGAMP alone or in combination with P12 (100 nM), CSE, and CSE-P12 (100 nM) for 24 h. The culture medium from each well was collected and centrifuged (14,000 rpm, 4 °C, 30 min) to precipitate the polypeptide gold nanoparticles; 20 μL of the supernatant was mixed with 180 μL of the QUANTIBlue solution in a 96-well plate and incubated at 37 °C until color development. The color change of the solution was evaluated by measuring the absorbance at 655 nm in a microplate reader, and an increase in the optical density indicated the activation of IRF or NF-κB / AP-1.

[0068] To investigate the effect of CSE-P12 on type I interferon effector factors produced by the cGAS-STING signaling pathway, THP-1 cells were seeded in 24-well culture plates and differentiated into macrophages with 50 ng / mL PMA; after 24 h, they were washed twice with PBS and left standing for 48 h before the experiment. Cells were treated with the STING agonist cGAMP alone or in combination with P12 (100 nM), CSE, and CSE-P12 (100 nM) for 24 h, the supernatant was discarded, and cell RNA was extracted by the Trizol method. The RNA concentration and purity were detected using Nanodrop to ensure that the OD260 / OD280 value was 1.8 - 2.0. The RNA of each sample was diluted to a uniform concentration according to the reverse transcription system. The expression of the MX1 gene was detected using RT-qPCR.

[0069] 2. Experimental Results

[0070] We used the THP-1 reporter cell system to evaluate the effect of CSE-P12 on the cGAS-STING downstream signaling pathways IRF and NF-κB / AP-1. The experimental results showed that CSE-P12 had a significant inhibitory effect on the activation of IRF and NF-κB / AP-1 induced by cGAMP in THP-1-derived macrophages ( Figure 1a and 1b ). By analyzing the expression of type I interferon effector factors using RT-qPCR, CSE-P12 could effectively inhibit the expression of the type I interferon-stimulated gene MX1 in cGAMP-stimulated THP-1-derived macrophages ( Figure 1c ).

[0071] Example 3

[0072] CSE-P12 Induces Autophagy and Degrades STING through Autophagy

[0073] 1. Experimental Methods

[0074] THP-1 cells were seeded in 24-well culture plates and differentiated into macrophages with 50 ng / mL PMA; after 24 h, they were washed twice with PBS and allowed to stand for 48 h before the experiment. Cells were treated with the STING agonist cGAMP alone or in combination with P12 (100 nM), CSE, or CSE-P12 (100 nM) for 4 h, or with the STING agonist cGAMP alone or in combination with Rapamycin (10 μM) and CSE-P12 (100 nM) for 4 h. The supernatant was discarded, and cell proteins were extracted. The expression of autophagy-related proteins p62, LC3II, and the key protein molecule STING was detected by Western blotting.

[0075] 2. Experimental Results

[0076] The expression levels of the key proteins p62 and LC3II for autophagosome formation were detected by Western blotting. We found that both CSE-P12 and P12 could upregulate the expression of p62 and LC3II, and CSE-P12 had a more significant effect on inducing autophagy ( Figure 2a ). To further verify the connection between CSE-P12-induced autophagy and STING, an autophagy inducer was used as a positive control. The results showed that both CSE-P12 and Rapamycin could degrade STING ( Figure 2b ).

[0077] Example 4

[0078] CSE-P12 Can Effectively Improve Adenovirus (HAdV4)-Induced Acute Pulmonary Inflammation 1. Experimental Methods

[0079] A mouse pneumonia model infected with adenovirus HAdV4 was constructed. Female C57BL / 6 wild-type mice (6 - 8 weeks old, SPF grade) were selected. First, the mice were anesthetized by intraperitoneal injection of 2.5% avertin. In the infection group, HAdV4 (5 × 10^6 PFU / Mouse, 20 μL) was administered by intranasal instillation, and in the administration group, CSE-P12 (500 nM, 50 μL) was administered by tracheal instillation through a laryngoscope 1 h before HAdV4 infection. The control group was treated with PBS in the same manner. On the 2nd and 4th days after infection, the drugs were administered continuously at the same dose and in the same manner. The experiment ended on the 5th day. The left lungs of the mice were collected and fixed with 4% paraformaldehyde for H&E staining. The right lung tissues were used for qPCR experiments to detect type I interferon cytokines. The histopathology of pulmonary inflammation was evaluated based on the following five features: alveolar neutrophils, interstitial neutrophils, alveolar hyaline membranes, protein exudation, and alveolar septal thickness.

[0080] 2. Experimental Results

[0081] Using an adenovirus (HAdV4)-induced mouse pneumonia model, we verified that CSE-P12 could effectively reduce the expression of the type I interferon gene Ifnb1 in lung tissues. By analyzing histopathological damage through H&E staining, it was confirmed that CSE-P12 could improve the pulmonary pathological damage caused by adenovirus infection, including the number of neutrophils in the alveolar cavity, the number of neutrophils in the lung interstitium, alveolar hyaline membrane formation, protein exudation, and alveolar septal thickening. Thus, CSE-P12 has a significant therapeutic effect on adenovirus (HAdV4)-induced pulmonary inflammation in mice ( Figures 3a - 3b ).

[0082] Example 5

[0083] CSE-P12 Can Effectively Treat Influenza Virus (PR8)-Induced Acute Pulmonary Inflammation

[0084] A mouse pneumonia model infected with influenza virus PR8 was constructed, and C57BL / 6 wild female mice (6 - 8 weeks old, SPF grade) were selected. The mice were anesthetized by intraperitoneal injection of 2.5% avertin. In the infection group, PR8 (5500 PFU / Mouse, 30 μL) was administered by nasal drip, and in the administration group, CSE-P12 (500 nM, 50 μL) was instilled into the trachea by laryngoscope 1 h before PR8 virus infection. The control group was treated with PBS in the same manner. On the 2nd, 4th, and 6th days, the drugs were continued to be administered at the same dose and in the same manner. The survival of mice in each group was observed continuously for 2 weeks, and the body weight changes of the mice were recorded daily. A mouse pneumonia model infected with influenza virus PR8 was constructed by the same modeling method as above. The experiment was terminated on the 6th day after infection, and the left lungs of the mice were collected and fixed with 4% paraformaldehyde for H&E staining.

[0085] 2. Experimental Results

[0086] In a mouse pneumonia model induced by influenza virus (PR8), we found that CSE-P12 could significantly improve the survival rate of PR8-infected mice. By analyzing histopathological damage through H&E staining, it was confirmed that CSE-P12 could improve the pulmonary inflammation and tissue damage caused by influenza virus infection. Thus, CSE-P12 has a significant therapeutic effect on influenza virus (PR8)-induced pulmonary inflammation in mice ( Figures 4a - 4b ).

[0087] Example 6

[0088] CSE-P12 Can Effectively Treat CLP-Induced Acute Lung Injury

[0089] To establish a mouse model of CLP-induced acute lung injury, C57BL / 6 wild male mice (6-8 weeks old, SPF grade) were selected. The body weight of the mice was weighed and recorded before the operation. The mice were placed in a vaporizer containing sevoflurane for several seconds. After the mice were motionless, gently shaking the vaporizer and the mice did not wake up, indicating successful anesthesia. The mice were fixed in the supine position on the operating board, marked and numbered at the tail. The abdominal surgical area was disinfected with iodophor. A longitudinal incision about 2 cm long was made along the midline of the abdomen, and the peritoneum was cut open to expose the abdominal cavity. The cecum was pulled out from the abdominal cavity with forceps, the direction of the mesenteric blood vessels was identified and the blood vessels were dissected. The cecum was ligated at 1 / 3 of the blind end with No. 3 silk thread, and a No. 7 injection needle was used to puncture the ligated end. The intestinal contents were gently squeezed out by pressing the intestinal tube, and the intestinal tube was replaced into the abdominal cavity. The abdomen of the mice was gently kneaded and sutured layer by layer to close the abdomen, and an appropriate amount of compound lidocaine cream was applied for pain relief. 1 mL of 0.9% sodium chloride injection was injected into the back of the mice for fluid replacement. After modeling, CSE-P12 (500 nM, 100 μL) was administered by intraperitoneal injection (ip). The mice were placed in an incubator to keep warm until they woke up. Then, CSE-P12 was continuously administered at the same dose at 24 h, 48 h, and 72 h after modeling for 7 consecutive days to observe the survival of mice in each group.

[0090] The mouse model of acute lung injury induced by CLP was established by the same modeling method as above. The experiment was terminated 72 h after modeling, and various organ tissues (whole blood, lung, liver, kidney, spleen) were collected after dissection. The left lung, the largest lobe of the liver, the left kidney, and half of the spleen of the mice were fixed in 4% neutral formaldehyde buffer for pathological analysis. The heart and the remaining lung, liver, kidney, and spleen tissues were frozen quickly in liquid nitrogen, ground thoroughly, and tissue homogenates were prepared with pre-cooled sterile PBS according to the ratio of body weight / volume (100 mg / mL). The homogenates were centrifuged at 2000 rpm for 10 min at 4°C, and the supernatants were taken. The obtained tissue supernatants were diluted at the original concentration or at a ratio of 1:10. Each time, 100 μL was taken and evenly spread on the LB solid medium, and cultured overnight at 37°C in a bacterial incubator. Then, the number of bacterial colonies (CFU) per mg of tissue was calculated to determine the severity of bacterial infection in each organ of the CLP model mice.

[0091] 2. Experimental Results

[0092] After CLP modeling, the survival rate of the mice was observed, and lung tissues were collected after the experiment for subsequent analysis of lung inflammation and injury. It was found that CSE-P12 could significantly improve the survival rate of CLP mice. The bacterial load in each organ of CLP mice was detected, and it was found that CSE-P12 could reduce the bacterial load in the spleen tissue of mice. The pathological sections of the lung tissues of CLP model mice stained with H&E were analyzed, and CSE-P12 could reduce the lung tissue injury and inflammatory response caused by sepsis bacterial infection ( Figures 5a - 5c ). The above results indicate that CSE-P12 can improve the survival rate of CLP mice and reduce the pathological injury of the lungs of mice caused by sepsis bacterial infection.

[0093] The foregoing has shown and described the basic principles, main features and advantages of the present invention. Each component mentioned in the present invention is a common technology in the existing art. Those skilled in the art should understand that the present invention is not limited by the above embodiments. The above embodiments and the descriptions in the specification only illustrate the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of the present invention claimed is defined by the appended claims and their equivalents.

Claims

1. A trace amount of cigarette extract CSE modified polypeptide gold nanoparticles, characterized in that The trace amount of CSE modifies its sequence SEQ ID NO: 1, the polypeptide gold nanoparticles are coded as CSE-P12, and the trace amount of CSE refers to 0.1%-1% cigarette extract.

2. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 1 or 2 to inhibit the cGAS-STING signaling pathway in vitro.

3. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 1 or 2 in inducing macrophage autophagy and degrading STING through autophagy.

4. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 1 or 2 in the treatment of acute lung inflammation induced by DNA viruses.

5. Use of a trace amount of CSE modified polypeptide gold nanoparticles according to claim 1 or 2 in the treatment of RNA virus-induced acute pulmonary inflammation, wherein the polypeptide gold nanoparticles are polypeptide-modified gold nanoparticles having a sequence as shown in SEQ ID NO:

1.

6. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 1 or 2 in a drug for treating sepsis-related acute lung injury.

7. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 4 in the treatment of acute lung inflammation induced by DNA viruses, wherein the DNA viruses are various adenoviruses.

8. Use of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of DNA virus-induced acute lung inflammation according to claim 7, wherein the adenovirus is human adenovirus HAdV4 or an adenovirus capable of infecting canine, porcine, ovine, or bovine mammals.

9. Use of a trace amount of CSE-modified polypeptide gold nanoparticles according to claim 5 in treating acute pulmonary inflammation induced by RNA viruses, wherein the RNA virus is influenza virus.

10. Use of a trace amount of CSE-modified polypeptide gold nanoparticles in the treatment of acute pulmonary inflammation induced by RNA viruses according to claim 9, wherein the influenza virus is PR8.

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