Application of PBX1 in preparation of medicine for preventing and treating acute lung injury

Abstract

The invention is applicable to the technical field of biological medicine, and provides application of PBX1 in preparation of medicine for preventing and treating acute lung injury, and the coding gene sequence of the PBX1 is as shown in SEQ ID No.1. The embodiment of the invention also provides a TAT-PBX1 recombinant protein and a virus for overexpressing the TAT-PBX1 recombinant protein. The invention further provides a medicine for preventing and treating the acute lung injury and a medical instrument for preventing and treating the acute lung injury, and the medicine is loaded on the medical instrument. According to the invention, the TAT-PBX1 recombinant protein is constructed, PBX1 can enter cells by virtue of the transmembrane effect of TAT to play a corresponding role in resisting oxidative damage, the TAT-PBX1 can relieve lung tissue pathological damage, inflammatory response and the like of ALI mice, and the TAT-PBX1 has a protective effect on LPS-induced inflammation.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the application of PBX1 in the preparation of drugs for preventing and treating acute lung injury. Background Art

[0002] As a tiny life form, viruses are widely present in nature, and as the natural environment changes, they constantly change their genetic and epigenetic information in an evolutionary way to adapt to environmental changes so that they can survive. As the scope of human social activities continues to increase, viruses that originally existed in biological organisms in the natural environment are very likely to invade the human body, causing serious harm to the population and even leading to the occurrence of public health emergencies. For example, acute and severe lung injury caused by acute respiratory viruses such as MERS, SARS, and COVID-19 has caused great harm to humans and seriously endangered human health. A series of antiviral drugs, such as ribavirin, acyclovir, ganciclovir, oseltamivir, etc., have been developed in the prior art to resist respiratory viral infections and have achieved certain clinical effects, but the efficacy of antiviral drugs depends largely on the type of virus and the specific condition of the patient. It is currently unknown whether these known antiviral drugs have a definite effect on unknown new respiratory viruses. The development of respiratory virus vaccines has always been a hot topic and focus of research in the field of biomedicine, but respiratory viruses mutate quickly, and the development of vaccines for unknown new respiratory viruses has a long cycle and is difficult to meet treatment needs. There is no way to say whether existing respiratory virus vaccines can produce an immune response to new respiratory viruses.

[0003] Given that acute and severe lung injury caused by acute respiratory viruses such as MERS, SARS, and COVID-19 share a common pathogenesis, that is, after the virus enters the respiratory tract, it induces an excessive immune response and severe inflammatory response in the body, causing large-scale necrosis in the infected person's lungs, and ultimately leading to death from respiratory failure. Therefore, how to inhibit inflammatory necrosis is a problem that needs to be solved in the prevention and treatment of acute lung injury. Summary of the invention

[0004] The purpose of the embodiments of the present invention is to provide the use of PBX1 in the preparation of drugs for preventing and treating acute lung injury, aiming to solve the problems raised in the above background technology.

[0005] The embodiment of the present invention is achieved by using PBX1 in the preparation of a drug for preventing and treating acute lung injury, wherein the coding gene sequence of the PBX1 is shown in SEQ ID No.1.

[0006] Another object of the embodiment of the present invention is to provide a TAT-PBX1 recombinant protein, the preparation of which includes the following steps: amplifying the coding gene of the PBX1 by PCR, connecting the PCR product to the cloning vector by cloning, and picking the correctly connected white clones by blue-white screening; after bacterial liquid PCR, enzyme digestion identification and gene sequencing identification, the PBX1 coding gene fragment in the positive clone is inserted into the prokaryotic expression vector pTAT-HA by enzyme digestion and T4 connection, and then transferred into the prokaryotic expression strain, and the positive transformant is obtained by ampicillin resistance screening; activating in the expression culture medium, expanding the culture, and collecting the precipitate by centrifugation; placing the precipitate in the lysis solution for ultrasonic crushing and dissolution, collecting the supernatant by centrifugation, filtering, purifying, and then renaturing by pulse dilution, dialyzing and ultrafiltration of the renaturing liquid to obtain the TAT-PBX1 recombinant protein.

[0007] Another object of an embodiment of the present invention is to provide a virus that overexpresses TAT-PBX1 recombinant protein, wherein the viral vector is one of a lentivirus, a retrovirus, an adenovirus, and an adeno-associated virus.

[0008] Another object of an embodiment of the present invention is to provide a drug for preventing and treating acute lung injury, wherein the drug comprises TAT-PBX1 recombinant protein or a virus overexpressing TAT-PBX1 recombinant protein.

[0009] Preferably, the medicine further comprises excipients.

[0010] Another object of an embodiment of the present invention is to provide a medical device for preventing and treating acute lung injury, wherein the medical device is loaded with the above-mentioned drug.

[0011] The use of PBX1 provided in an embodiment of the present invention in the preparation of a drug for preventing and treating acute lung injury, the pre-B-cell leukemia homeobox 1 (PBX1) is a member of the homeobox gene family, which encodes the transcription factor PBX1, which is widely expressed in a variety of tissues, and its functions include self-renewal, proliferation and differentiation of stem cells. PBX1 not only plays an important role in the biological development process, but also has a regulatory effect on many organs. In terms of nerves, PBX1 participates in neurogenesis and is a promoter of neurogenesis in the subventricular zone of the adult ventricle; in the development of NK cells, PBX1 plays a key role by directly binding to the Nfil3 promoter; PBX1 can also promote proliferation and inhibit cell apoptosis, thereby inducing abnormal cardiac development. It is essential in bone formation, the development of the adrenal glands, pancreas, urogenital organs, kidney generation and maintenance of hematopoiesis; however, as a transcription factor, PBX1 cannot be expressed by cells due to its large molecular weight. The cell membrane enters the cell. The embodiment of the present invention constructs the TAT-PBX1 recombinant protein. PBX1 can enter the cell with the help of the transmembrane effect of TAT, and exerts a corresponding antioxidant damage effect. The acute lung injury mouse model is caused by tracheal instillation of lipopolysaccharide (LPS), simulating severe lung injury induced by an unknown new acute respiratory virus, and observing the effects of TAT-PBX1 on reducing lung tissue pathological damage and inflammatory response in acute lung injury (ALI) mice; LPS is used to stimulate human alveolar epithelial cells A549 to create an in vitro model, and the protective effect of TAT-PBX1 on LPS-induced inflammation is observed. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 The plasmid map of TAT-PBX1 provided in the embodiment of the present invention;

[0013] Figure 2 a is the construction of the TAT-PBX1 prokaryotic expression system provided in the embodiment of the present invention and the denaturation, purification and renaturation results of the recombinant protein; Figure 2 b: Three hours after intraperitoneal injection of TAT-PBX1 (2.5 mg / Kg), His-tag was detected in the lung tissue of mice, indicating that TAT-PBX1 can enter the lung tissue.

[0014] Figure 3 a, b, and c are the results of the mouse body weight, organ index, and lung tissue wet-to-dry weight ratio provided in the examples of the present invention, respectively (CON is the control group; LPS is the model group; before-L, before-M, and before-H are the low, medium, and high dose groups of TAT-PBX1 injected intraperitoneally before LPS induction; after-L, after-M, and after-H are the low, medium, and high dose groups of TAT-PBX1 injected intraperitoneally after LPS induction);

[0015] Figure 4 HE staining results of mouse lung tissue provided in the embodiment of the present invention;

[0016] Figure 5 a is the protein concentration test result of bronchoalveolar lavage fluid of each group of mice provided in the embodiment of the present invention, Figure 5 b is the total cell count results of bronchoalveolar lavage fluid of mice in each group.

[0017] Figure 6 a is the detection result of inflammatory factors in bronchoalveolar lavage fluid of each group of mice provided in the embodiment of the present invention, Figure 6 b is the detection results of inflammatory factors in the serum of mice in each group.

[0018] Figure 7 The ELISA provided in the embodiment of the present invention detects the inflammatory factors of human alveolar epithelial cells;

[0019] Figure 8 The flow cytometry provided in the embodiment of the present invention is used to detect the ROS situation of human alveolar epithelial cells;

[0020] Fig. 9 The flow cytometry provided in the embodiment of the present invention is used to detect the mitochondrial membrane potential of human alveolar epithelial cells;

[0021] Fig.10 The Western Blot protein detection results of mouse lung tissue provided in the embodiments of the present invention. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0023] The specific implementation of the present invention is described in detail below in conjunction with specific embodiments.

[0024] Example 1: Preparation of TAT-PBX1 recombinant protein, comprising the following steps:

[0025] The coding gene of PBX1 (its sequence is shown in SEQ ID No. 1) was amplified by PCR, and the PCR product was connected to the cloning vector pT-EASY by TA cloning, and then the correctly connected white clone was picked by blue-white screening; after bacterial liquid PCR, restriction digestion identification and gene sequencing identification, the PBX1 coding gene fragment in the positive clone was inserted into the prokaryotic expression vector pTAT-HA by restriction digestion and T4 connection, and then transferred into the prokaryotic expression strain Rosetta (DE3), and the positive transformant was obtained by ampicillin resistance screening. Activate overnight at 37°C in the expression medium, expand the culture, and collect the precipitate by centrifugation; the precipitate was placed in the lysis solution for ultrasonic crushing and dissolution, centrifuged at 12000g×30min, and the supernatant was collected; the supernatant was filtered with a 0.45μm filter, purified by nickel column affinity chromatography, and then renatured by pulse dilution, and the renatured solution was dialyzed and ultrafiltered to obtain a high concentration of TAT-PBX1 recombinant protein. The plasmid map of TAT-PBX1 is shown in Figure 1 As shown, the DNA sequence of PBX1 was added to the C-terminus of the TAT peptide within the plasmid vector pTAT-HA, which contained an N-terminal His 6 tag for purification and an HA tag for distinguishing TAT from PBX1;

[0026] The specific purification steps of the TAT-PBX1 recombinant protein are as follows: using a Ni2+-NTA-agarose column, after equilibration with a binding buffer, loading the bacterial supernatant onto the column, washing the column with a washing buffer, and then eluting the TAT-PBX1 recombinant protein bound to the column with an elution buffer;

[0027] The specific renaturation steps of the TAT-PBX1 recombinant protein are as follows: the eluted sample is renatured in a renaturation buffer at a dilution ratio of 1:1-20. After the renaturation is completed, the supernatant is introduced into a dialysis bag and dialyzed with phosphate buffer at 4°C overnight. After that, the denatured protein is concentrated using a 30-kDa ultrafiltration tube, and the concentration of the recombinant protein is determined using a BCA protein detection kit.

[0028] Example 2: Experiment on TAT-PBX1 alleviating acute lung injury:

[0029] In vitro experiments: LPS was used to model the cells and the cells were divided into blank control group, TAT-PBX1 recombinant protein group, LPS group, and LPS+TAT-PBX1 recombinant protein group. ELISA kits were used to detect inflammatory factors in cell supernatants, DCFH-DA probes were used to detect cell ROS, and JC-1 was used to detect cell mitochondrial membrane potential.

[0030] In vivo experiment: 6-8 week old male SPF grade C57BL / 6 mice were raised at a constant temperature of 22℃ and constant humidity in a quiet environment. The mice were randomly divided into 5 groups, namely: blank control group, LPS group, TAT-PBX1 recombinant protein low-dose group, TAT-PBX1 recombinant protein medium-dose group, and TAT-PBX1 recombinant protein high-dose group; low (2.5 mg / kg), medium (5 mg / kg), and high (10 mg / kg) doses of TAT-PBX1 recombinant protein were intraperitoneally injected into the mice, respectively, and LPS was instilled into the trachea 3 hours later to establish the model; 24 hours later, the mice were killed by cervical dislocation, and the required organs and samples were taken for pathological observation, ELISA inflammatory factor detection, bronchoalveolar lavage fluid protein concentration and total cell number detection, flow cytometry detection of lung tissue ROS and mitochondrial membrane potential, and Western blot detection of related protein expression;

[0031] Statistical analysis: All data are expressed as mean ± standard deviation. Statistical analysis and statistical graph generation were performed using GraphPad Prism 8 software. When comparing between groups, if the data conformed to normal distribution, one-way ANOVA test was used, and the least significant difference test (LSID) was used for multiple comparisons between groups; if the data variance was unequal, the Kruskal-Wallis test was used, and the test level was a-0.05. p<0.05 was considered statistically significant, and p<0.01 was considered statistically significant.

[0032] The construction of TAT-PBX1 prokaryotic expression system and the results of denaturation, purification and renaturation of recombinant protein are shown in Figure 2 As shown in a, according to the Coomassie Brilliant Blue results of the bacterial solution (also called the starting solution, Start), the flowthrough solution (Flowthrough), the washing solution (Wash) and the elution solution (Elution), it is proved that the purity of the purified TAT-PBX1 is greater than 90%, the molecular weight is correct, and it can be successfully detected; Figure 2 b: Three hours after intraperitoneal injection of TAT-PBX1 (2.5 mg / Kg), His-tag was detected in the lung tissue of mice, indicating that TAT-PBX1 can enter the lung tissue.

[0033] Result analysis:

[0034] TAT-PBX1 recombinant protein alleviates LPS-induced acute lung injury in mice:

[0035] First, the effect of TAT-PBX1 recombinant protein was evaluated by observing the differences in body weight, organ index and lung tissue wet-to-dry weight ratio of each group of mice. Figure 3 As shown, Figure 3a is the weight of mice in each group. Compared with the control group, the weight of mice in the LPS group was significantly reduced, while the weight of mice in the before-H group was significantly increased; Figure 3 b is the lung tissue organ index of mice in each group. Compared with the control group, the lung tissue organ index of mice in the LPS group was significantly increased, while the lung index of mice in the before-H group was significantly decreased; Figure 3 c is the wet-to-dry weight ratio of lung tissue of mice in each group. Compared with the control group, the wet-to-dry weight ratio of lung tissue of mice in the LPS group was significantly increased, while the lung index of mice in the before-M, before-H and after-H groups was significantly decreased. The results of the lung tissue organ index and wet-to-dry weight ratio indicate that LPS causes a certain degree of lung tissue edema in mice, and TAT-PBX1 recombinant protein can alleviate lung edema.

[0036] Through the observation of pathological sections, we explored the effect of TAT-PBX1 recombinant protein on the recovery of LPS-induced lung tissue damage in mice. Figure 4 The lung tissue pathology of mice with acute lung injury induced by LPS was observed before and after intraperitoneal injection of TAT-PBX1 recombinant protein. The results of liver tissue HE staining were observed under an optical microscope. The alveolar walls of mice in the control group were thinner, with only a small amount of inflammatory cells infiltrating in the alveolar cavity, no bleeding spots, and intact lung tissue structure; while the alveolar walls of LPS mice were thickened, accompanied by a large number of inflammatory cells infiltrating, and the lung tissue structure was severely damaged; whether TAT-PBX1 was injected first or later, the alveolar cell structure was significantly restored, the inflammatory cell infiltration was significantly reduced, and the alveolar wall became thinner;

[0037] The permeability of mouse lung tissue was evaluated by detecting the protein concentration and total cell number of mouse bronchoalveolar lavage fluid. The results were as follows: Figure 5 As shown, Figure 5 a is the protein concentration test results of bronchoalveolar lavage fluid of mice in each group. Compared with the control group, the protein concentration of bronchoalveolar lavage fluid of mice in the LPS group was significantly increased, while the protein concentration of bronchoalveolar lavage fluid of mice in the before-H group was significantly decreased; Figure 5 b is the total cell count results in the alveolar lavage fluid of mice in each group. Compared with the control group, the total cell number in the alveolar lavage fluid of mice in the LPS group increased significantly, while the total cell number in the alveolar lavage fluid of mice in the before-H group decreased significantly. The above results indicate that LPS causes increased permeability of mouse lung tissue and dysfunction of alveolar capillary barrier, while TAT-PBX1 recombinant protein can alleviate lung damage;

[0038] The ELISA method was used to detect the levels of inflammatory factors in the mouse bronchoalveolar lavage fluid and serum to evaluate the effect of TAT-PBX1 recombinant protein on inflammatory response. Figure 6 As shown, Figure 6a is the detection results of inflammatory factors in the bronchoalveolar lavage fluid of mice in each group. Compared with the control group, the levels of inflammatory factors in the bronchoalveolar lavage fluid of mice in the LPS group were significantly increased, while those in the bronchoalveolar lavage fluid of mice in the before-H group were significantly decreased; Figure 6 b is the detection results of inflammatory factors in the serum of mice in each group. Compared with the control group, the level of inflammatory factors in the serum of mice in the LPS group was significantly increased, while the level of inflammatory factors in the serum of mice in the before-H group was significantly decreased. The above results indicate that LPS caused inflammatory response in the lung tissue and systemic of mice, while TAT-PBX1 recombinant protein can reduce the inflammation in the lung tissue and systemic of mice.

[0039] TAT-PBX1 recombinant protein can reverse LPS-stimulated inflammation, ROS and changes in mitochondrial membrane potential in human alveolar epithelial cells:

[0040] An in vitro model of acute lung injury was established by stimulating human type II alveolar epithelial cells A549 with LPS. Inflammatory factors were detected by ELISA, and ROS and mitochondrial membrane potential of human alveolar epithelial cells were detected by flow cytometry to evaluate whether TAT-PBX1 recombinant protein could reverse LPS-induced changes in human alveolar epithelial cell inflammation, ROS and mitochondrial membrane potential.

[0041] Figure 7 The results of ELISA detection of inflammatory factors showed that compared with the control group, the inflammatory factors in the LPS group increased significantly, while those in the before-H group decreased significantly, indicating that the TAT-PBX1 recombinant protein can alleviate the inflammatory response; Figure 8 As for the ROS situation of human alveolar epithelial cells, compared with the control group, the ROS in the LPS group increased significantly, while the ROS in the before-H group decreased significantly, indicating that TAT-PBX1 recombinant protein can reduce ROS; Fig. 9 As for the mitochondrial membrane potential of human alveolar epithelial cells, compared with the control group, the mitochondrial membrane potential of the LPS group was significantly decreased, while the mitochondrial membrane potential of the before-H group was significantly increased, indicating that the TAT-PBX1 recombinant protein can restore the mitochondrial membrane potential.

[0042] TAT-PBX1 recombinant protein reduces apoptosis in mouse lung tissue:

[0043] The expression of apoptosis-related proteins was detected by Western Blot, and the results were as follows Fig.10As shown in the figure, compared with the control group, the Cleaved Caspase3 in the LPS group was significantly increased, and the difference was statistically significant (p < 0.05). The expression levels of apoptosis-related proteins in the low, medium and high dose groups of TAT-PBX1 were significantly downregulated compared with the LPS group. Compared with the control group, the γH2A.X in the LPS group was significantly increased, and TAT-PBX1 significantly downregulated the expression level of γH2A.X protein, and the difference was statistically significant (p < 0.05). In addition, TAT-PBX1 can significantly upregulate the levels of AMPKγ2, PGC-1α and TFAM proteins (p < 0.05), indicating that PBX1 may promote mitochondrial biogenesis; it can be seen that the TAT-PBX1 recombinant protein may reduce LPS-induced lung tissue DNA damage and apoptosis by regulating the AMPKγ2 / PGC-1α / TFAM pathway.

[0044] In summary, PBX1 slows down cell aging and inhibits apoptosis by reducing the accumulation of reactive oxygen species and inhibiting mitochondrial damage mediated by inflammation and excessive immune response. Cell-penetrating peptides (CPPs) have been used to deliver a variety of molecules, especially large molecular weight active proteins from the extracellular to the intracellular, such as protamine, TAT peptide or R9, among which transcription transactivator (TAT) is a peptide derived from human immunodeficiency virus and is one of the most commonly used cell-penetrating peptides. When TAT binds to cells, it can enhance the cell's absorption of proteins.

[0045] The PBX1 provided in the embodiment of the present invention is present in the culture medium of cells (such as mesenchymal stem cells, neural crest stem cells, pigment stem cells, and human cells such as fibroblasts). In order to facilitate membrane permeation, PBX1 generally further includes a membrane-penetrating peptide, such as TAT;

[0046] The TAT-PBX1 overexpressing cells or culture supernatant provided in the embodiment of the present invention are cells infected with lentivirus to overexpress TAT-PBX1 protein, because the TAT membrane-penetrating peptide enables the PBX1 protein contained in the cell culture supernatant to enter the cell;

[0047] The TAT-PBX1 recombinant protein provided in the embodiment of the present invention is an exogenous protein expressed by Escherichia coli, yeast or CHO cells. Since TAT cell-penetrating peptide enhances the absorption of protein by alveolar epithelial cells, neurons, mesenchymal stem cells and other cells, PBX1 can pass through the cell membrane and enter the cell nucleus. Experimental results show that the prokaryotically expressed TAT-PBX1 still has transcriptional activity after entering the cell nucleus.

[0048] The TAT-PBX1 recombinant protein provided in the embodiment of the present invention is packaged with lentivirus, adenovirus, adeno-associated virus and retrovirus, and then transduced into human cells, including but not limited to mesenchymal stem cells, neural crest stem cells, pigment stem cells and fibroblasts;

[0049] Upregulation of PBX1 can significantly reduce LPS-induced excessive accumulation of intracellular ROS, mitochondrial damage and cell apoptosis. Infection of mesenchymal stem cells with lentivirus overexpressing TAT-PBX1 can upregulate the expression of PBX1 protein.

[0050] TAT-PBX1 recombinant protein entered the lung and other tissues of mice after intraperitoneal injection or gastrointestinal administration;

[0051] Using PBX1 or TAT-PBX1 as an active ingredient for preventing and treating acute lung injury, specifically, mixing PBX1 or TAT-PBX1 with other excipients to prepare a medicine (preparation) for preventing and treating acute lung injury;

[0052] Pharmaceutical excipients may be those conventionally used in various preparations, such as but not limited to isotonic agents, buffers, flavoring agents, excipients, fillers, adhesives, disintegrants and lubricants, etc.; they may also be selected for use in order to be compatible with the substance, such as emulsifiers, solubilizers, antibacterial agents, analgesics and antioxidants, etc. Such excipients can effectively improve the stability and solubility of the compounds contained in the composition or change the release rate and absorption rate of the compounds, thereby improving the metabolism of various compounds in the organism, and further enhancing the administration effect of the composition;

[0053] In aqueous solution injections, excipients generally include isotonic agents and buffers, as well as necessary emulsifiers (such as Tweeen-80, Pluronic and Poloxamer), solubilizers and antibacterial agents, etc. In addition, other pharmaceutically acceptable excipients are also included, such as antioxidants, pH regulators and analgesics, etc.

[0054] The excipients used to prepare liquid preparations generally include solvents, water, oils (such as fatty acids), emulsifiers, and necessary preservatives;

[0055] Various excipients and the culture medium supernatant prepared in the embodiments of the present invention are made into dosage forms that are conducive to drug delivery, such as but not limited to aqueous solution injections, nasal drops, eye drops, powder injections, powders, patches, suppositories, emulsions, creams, gels, aerosols, sprays, powder aerosols, sustained-release agents and controlled-release agents. In addition, excipients such as but not limited to gelatin, albumin, chitosan, polyether and polyester polymer materials, such as but not limited to polyethylene glycol, polyurethane, polycarbonate and its copolymers, etc., can be used to achieve specific drug delivery purposes or methods, such as sustained-release drug delivery, controlled-release drug delivery and pulse drug delivery. The main manifestations of the so-called "conducive to drug delivery" include but are not limited to improving therapeutic effects, improving bioavailability, reducing toxic side effects and improving patient compliance, etc.;

[0056] In addition, drugs and medical devices can also be combined to make medicated medical devices, such as dressings containing TAT-PBX1 according to the embodiments of the present invention, where TAT-PBX1 according to the embodiments of the present invention is loaded or coated on the material as an active ingredient to make medical devices for preventing and treating acute lung injury. Common scaffold materials such as PLA, PLGA, GelMA and metals, as well as microneedles and microneedle arrays thereof mixed with biocompatible degradable materials, or loaded in metal microneedles to make microneedle chips. When the microneedles penetrate the skin, TAT-PBX1 is released in the epithelial tissue, enters the blood, reaches the damaged organ, and prevents and treats acute lung injury.

[0057] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. The use of PBX1 in the preparation of a drug for preventing and treating acute lung injury, characterized in that: The coding gene sequence of PBX1 is shown in SEQ ID No.

1.

2. A TAT-PBX1 recombinant protein, characterized in that: The preparation of the recombinant protein comprises the following steps: The PBX1 coding gene as claimed in claim 1 is amplified by PCR, the PCR product is connected to the cloning vector by cloning, and the correctly connected white clone is picked by blue-white screening; after bacterial liquid PCR, enzyme digestion identification and gene sequencing identification, the PBX1 coding gene fragment in the positive clone is inserted into the prokaryotic expression vector pTAT-HA by enzyme digestion and T4 connection, and then transferred into the prokaryotic expression strain, and the positive transformant is obtained by ampicillin resistance screening; activation in expression culture medium, expansion culture, centrifugation to collect precipitation; placing the precipitation in a lysis solution for ultrasonic crushing and dissolution, centrifugation to collect the supernatant, filtering, purification, and then renaturation by pulse dilution, dialysis and ultrafiltration of the renaturation solution to obtain the TAT-PBX1 recombinant protein.

3. A virus overexpressing TAT-PBX1 recombinant protein, characterized in that: The viral vector is one of a lentivirus, a retrovirus, an adenovirus, and an adeno-associated virus.

4. A drug for preventing and treating acute lung injury, characterized in that: The drug comprises the TAT-PBX1 recombinant protein as claimed in claim 2 or the TAT-PBX1 recombinant protein overexpressing virus as claimed in claim 3.

5. The drug according to claim 4, characterized in that The medicine also includes excipients.

6. A medical device for preventing and treating acute lung injury, characterized in that: The medical device is loaded with the medicine as claimed in claim 4.

Images