Use of follicle stimulating hormone in preparation of medicine for preventing and treating sepsis acute lung injury

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to application of follicle stimulating hormone in preparation of a medicine for preventing and treating sepsis acute lung injury. The application adopts a clinically relevant sepsis acute lung injury model, i.e. a LPS-induced mouse sepsis acute lung injury model and a human lung microvascular endothelial cell inflammation model. Experiments prove that intraperitoneal injection of follicle stimulating hormone can significantly inhibit LPS-induced mouse sepsis acute lung injury, improve the survival rate, reduce lung edema, bleeding and inflammatory cell infiltration, and reduce the lung injury pathological score; and can inhibit endothelial cell activation and stabilize endothelial barrier function, and can be used for prevention and treatment of sepsis acute lung injury.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of follicle-stimulating hormone in the preparation of drugs for the prevention and treatment of acute lung injury in sepsis. Background Technology

[0002] Sepsis is a clinical syndrome caused by immune-inflammatory homeostasis imbalance due to infection or injury. The lungs are the most susceptible target organ for sepsis, with over 50% of sepsis patients developing acute lung injury (ALI) or even acute respiratory distress syndrome (ADRS). ADRS manifests clinically as refractory hypoxemia, non-cardiogenic pulmonary edema, and acute respiratory failure, essentially due to the disruption of the pulmonary microvascular endothelial barrier caused by systemic inflammatory response. Therefore, inhibiting pulmonary microvascular endothelial cell activation is a key step in stabilizing the pulmonary microvascular endothelial barrier and holds promise for providing a new strategy for early intervention in sepsis-induced acute lung injury.

[0003] However, current clinical practice mainly relies on symptomatic support measures such as anti-infection and respiratory support, and there is a lack of specific drugs to stabilize the pulmonary microvascular endothelial barrier and thus treat acute lung injury caused by sepsis. Follicle-stimulating hormone (FSH) is a glycoprotein secreted by the pituitary gland, whose main function is to regulate follicle development and estrogen synthesis. Recombinant human FSH has become a first-line ovulation-inducing drug in the field of assisted reproduction. Currently, there are no reports on the effects of FSH on sepsis. Summary of the Invention

[0004] This invention addresses the treatment of acute lung injury in sepsis, aiming to provide the application of follicle-stimulating hormone (FSH) in the preparation of drugs for the prevention and treatment of acute lung injury in sepsis. This invention discovers that FSH protects the pulmonary microvascular endothelial barrier by inhibiting the activation of pulmonary microvascular endothelial cells (PMVECs), thereby improving sepsis and sepsis-induced acute lung injury.

[0005] The research process of this invention is as follows: First, a septic mouse model was established by intraperitoneal injection of lipopolysaccharides (LPS) to observe the protective effect of FSH on acute lung injury caused by sepsis; then, an inflammation model was established by stimulating PMVECs with LPS, and in vitro experiments were performed using Western blot, immunofluorescence staining, and RT-qPCR. In vivo animal experiments showed that FSH treatment could significantly improve the survival rate of septic mice, reduce pulmonary edema, hemorrhage, and inflammatory cell infiltration, and reduce the pathological score of lung injury; in vitro cell experiments showed that after FSH acted on PMVECs, it could reduce the expression of the endothelial activation marker ICAM1, improve the continuity of intercellular connections, and downregulate inflammatory genes. IL1B and IL6 The expression level indicates that FSH improves acute lung injury caused by sepsis by inhibiting PMVEC activation and stabilizing the pulmonary microvascular endothelial barrier.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows: In a first aspect, the invention provides the use of follicle-stimulating hormone in the preparation of a medicament for the prevention and / or treatment of acute lung injury caused by sepsis in a subject.

[0007] Furthermore, the acute lung injury caused by sepsis is an acute complication of sepsis, which is caused by bacterial infection.

[0008] In the highly inflammatory environment of sepsis, PMVECs, stimulated by bacterial endotoxins, oxygen free radicals, and inflammatory mediators secreted by inflammatory cells, rapidly transition from a resting state to an activated state characterized by pro-apoptosis, pro-inflammation, procoagulation, and high adhesion. This leads to increased vascular permeability, increased transendothelial migration of inflammatory cells, and exudation of erythrocytes and proteins, resulting in tissue edema and impaired lung function. In particular, when sepsis is caused by bacterial infection, toxic substances such as LPS released by bacteria can exacerbate multi-organ damage, intensifying the symptoms and treatment difficulty of acute lung injury in sepsis. Activated PMVECs, on the one hand, secrete inflammatory mediators that directly exacerbate their own damage; on the other hand, they secrete chemokines to recruit more inflammatory cells, forming a malignant cascade reaction that ultimately leads to the destruction of the pulmonary microvascular barrier.

[0009] This invention, through in vitro and in vivo experiments, reveals that FSH can directly target the FSH receptor expressed on PMVECs, inhibiting the overactivation of PMVECs under inflammatory stimuli such as LPS. Specifically, FSH can reduce the expression of the endothelial activation marker ICAM1, alleviate the disruption of intercellular connections, and downregulate inflammatory genes. IL1B and IL6FSH treatment significantly improved endothelial barrier function, reduced vascular leakage, and decreased inflammatory cell infiltration by lowering FSH expression levels. In a mouse model of sepsis, FSH treatment significantly improved survival rates, reduced pulmonary edema, hemorrhage, and alveolar structural damage, and lowered lung injury pathology scores. Therefore, FSH effectively prevents and treats acute lung injury caused by sepsis by stabilizing the pulmonary microvascular endothelial barrier.

[0010] The beneficial effects of this invention are: This invention is the first to propose the application of follicle-stimulating hormone (FSH) in the preparation of drugs for the prevention and treatment of acute lung injury caused by sepsis. In vivo animal experiments confirmed that FSH significantly improves the survival rate of septic mice, reduces pulmonary edema, hemorrhage, and inflammatory cell infiltration, lowers the pathological score of lung injury, and improves overall prognosis. In vitro cell experiments confirmed that FSH can reduce the expression of the activation marker ICAM1 in PMVECs, improve intercellular junction integrity, and downregulate inflammatory genes. IL1B and IL6 The expression of follicle-stimulating hormone (FSH) inhibits the activation of pulmonary microvascular endothelial cells, thereby stabilizing endothelial barrier function. Therefore, this invention demonstrates the efficacy of FSH in the preparation of drugs for the prevention and treatment of acute lung injury caused by sepsis, providing new evidence for the treatment of acute lung injury caused by sepsis. Attached Figure Description

[0011] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0012] Figure 1 Survival curves showing how FSH improves the survival rate of septic mice; Figure 2 Lung tissue images for FSH-mediated lung injury in septic mice (scale bar = 2 mm). Figure 3 A statistical graph showing the reduction of the wet-to-dry weight ratio of lung tissue in septic mice by FSH. Figure 4 Lung tissue sections and H&E staining images for FSH-mediated lung injury in septic mice; Figure 5 Statistical graph showing how FSH reduces lung injury pathological scores in septic mice; Figure 6 Figure 1. Western blot results confirming FSH receptor expression in human PMVECs; Figure 7 Immunofluorescence staining results for FSH to improve endothelial barrier damage (arrows indicate the interruption of intercellular connections); Figure 8 To reduce the inflammatory genes of PMVECs by FSH IL1B and IL6 A statistical graph of relative expression levels, in which Figure 8 A is IL1B Statistical graph of relative expression levels Figure 8 B is IL6 Statistical graph of relative expression levels; Figure 9 Western blot results for FSH-induced reduction of endothelial activation marker ICAM1 expression; in, P <0.001, P <0.01, P <0.05. Detailed Implementation

[0013] Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be construed as limiting the scope of the invention. Specific conditions not specified in the embodiments are performed under conventional conditions or conditions recommended by the manufacturer. Components whose manufacturers are not specified are all commercially available conventional products.

[0014] In this invention, "and / or" includes any and all combinations of one or more of the listed related items.

[0015] In this invention, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0016] It should be noted that, in this invention, the terms "comprising," "including," or any other variations thereof are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0017] As used in this specification, the terms “about” or “approximately” typically mean + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, even more typically + / -0.5% of the value.

[0018] In this specification, some embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and the individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0019] This invention provides the use of follicle-stimulating hormone in the preparation of a medicament for the prevention and / or treatment of acute lung injury caused by sepsis in a subject.

[0020] The terms "follicle-stimulating hormone" and "FSH" are used interchangeably, both referring to a class of hormones or variants thereof used to promote sperm or follicle production and ovarian development. They are naturally secreted by the anterior pituitary gland and can also be extracted from the urine of postmenopausal women or obtained through recombinant technology. The FSH used in this invention can be obtained by any method commonly used in the art, such as naturally occurring, obtained through recombinant technology, or synthesized. In one embodiment of this invention, the follicle-stimulating hormone is human follicle-stimulating hormone or a variant thereof, including but not limited to follicle-stimulating hormone derived from human urine or recombinant human follicle-stimulating hormone, as well as high-purity human menopausal gonadotropins, without specific limitations herein. Before purifying FSH using the method of this invention, the raw material FSH can be preliminarily purified using conventional methods in the art.

[0021] In some embodiments, the acute lung injury caused by sepsis is an acute complication of sepsis, which is caused by bacterial infection; specifically, the bacteria are Gram-negative bacteria; more specifically, the acute lung injury caused by sepsis is LPS-induced acute lung injury in mice with sepsis.

[0022] In some embodiments, sepsis can be caused by pathogens such as fungi or viruses in addition to bacterial infection; the source of infection for sepsis is not limited here.

[0023] In some embodiments, the drug may be introduced into the body, such as into muscles, intradermal tissues, subcutaneous tissues, veins, or mucous membranes, via injection, oral administration, spraying, penetration, absorption, or physical or chemical means; or it may be introduced into the body after being mixed with or encapsulated by other substances. In some embodiments, the drug may be administered via intraperitoneal injection, intramuscular injection, intravenous injection, or oral administration; further, the drug may be administered via intraperitoneal injection.

[0024] In some embodiments, the single dose of the drug, calculated based on follicle-stimulating hormone (FSH), is 15-45 IU / kg; further, the single dose of the drug, calculated based on FSH, is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, or 45 IU / kg; and even further, the single dose of the drug, calculated based on FSH, is 30 IU / kg.

[0025] In some embodiments, the medicament may also include a pharmaceutically acceptable carrier and / or adjuvant.

[0026] In some embodiments, the drug is a unit dosage form of a solid or liquid preparation; the dosage form includes capsules, pills, tablets, powders, granules, lozenges, lozenges, melts, powders, solutions, suspensions, or emulsions. Drugs in all of the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0027] In some embodiments, the solid or liquid formulation is provided in individual dosage units, thereby providing a predetermined amount of active ingredient upon administration.

[0028] As used in this invention, the term "pharmaceutical acceptable" means a compound, raw material, composition, and / or dosage form that, within a reasonable scope of medical judgment, is suitable for contact with a subject's tissues without excessive toxicity, irritation, allergic reactions, or other problems and complications exceeding a reasonable benefit / risk ratio, and is effective for its intended use.

[0029] In some embodiments, the pharmaceutically acceptable carriers and / or excipients of the present invention include, but are not limited to, solvents, diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorbent carriers, lubricants, etc., which are conventional in the pharmaceutical field and are added at appropriate steps in the preparation process.

[0030] In some embodiments, the drug is prepared in the form of a dose unit, configured as an independent dose for single use containing 900-2700 IU of follicle-stimulating hormone, or configured as a dose unit in at least one independent dose form; further, the drug is prepared in the form of a dose unit, configured as an independent dose for single use containing 1500-2100 IU of follicle-stimulating hormone, or configured as a dose unit in at least one independent dose form; even further, the drug is prepared in the form of a dose unit, configured as an independent dose for single use containing 1800 IU of follicle-stimulating hormone, or configured as a dose unit in at least one independent dose form.

[0031] In some embodiments, the drug is directly targeted to the lungs of the subject and taken up by the subject's lung tissue.

[0032] In some embodiments, the symptoms of acute lung injury caused by sepsis include one or more of the following: localized bleeding in the lung tissue, perialveolar inflammation, and alveolar inflammatory lesions.

[0033] In some embodiments, the drug is a drug that improves vascular endothelial cell function under sepsis conditions or improves the pathological condition of acute lung injury in sepsis.

[0034] In some embodiments, the drug is used to reduce the inflammatory response in the lungs of the subject.

[0035] As used in this invention, "subject" means a subject who needs treatment for a disease, and more specifically, means a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse or cow, but the invention is not limited thereto.

[0036] In some embodiments, the drug for improving vascular endothelial cell function under sepsis conditions is a drug that improves endothelial cell barrier function, reduces monocyte adhesion, or lowers the level of cell inflammation; the drug for improving the pathological condition of acute lung injury in sepsis includes drugs that improve the pathological structure of lung tissue, reduce pulmonary edema and tissue protein exudation, or improve the pulmonary vascular barrier capacity.

[0037] In some embodiments, the inflammation-related genes include, but are not limited to, those mentioned above. IL1B , IL2 , IL4 , IL6 , IL17A , IL18 , IL23A , IFNF , TNFA , TGFB1 In a specific embodiment of the present invention, the inflammation-related gene is... IL1B IL6 .

[0038] Unless otherwise specified, the experimental methods described in the following examples are conventional methods.

[0039] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0040] All percentages mentioned in the following examples are mass percentages, unless otherwise stated.

[0041] This invention discloses the application of follicle-stimulating hormone (FSH) in the preparation of drugs for the prevention and treatment of acute lung injury in sepsis, and conducts in vitro cell experiments and in vivo animal experiments, as detailed below: Reagent sources: FSH for animal experiments was purchased from MERCK Biotech, and FSH for cell experiments was purchased from R&D Systems; LPS was purchased from MERCK Biotech; RNA extraction kit for RT-qPCR was purchased from Shanghai Yishan Biotechnology Co., Ltd.; SYBR Green qPCR premix was purchased from Aibote Biotechnology Co., Ltd.; primers were purchased from Shanghai Beyotime Biotechnology Co., Ltd.; follicle-stimulating hormone receptor (FSH receptor, FSHR) antibody was purchased from Wuhan Sanying Biotechnology Co., Ltd.; VE-cadherin antibody was purchased from Immunoway, which is an antibody used to detect vascular endothelial cadherin (VE-cadherin), also known as CD144 antibody or Cadherin-5 antibody; ICAM1 antibody was purchased from Immunoway.

[0042] Statistical methods: First, the Shapiro-Wilk test was used to test the normality of the data. For normally distributed data with homogeneous variances, independent samples were used between the two groups. t For comparisons among multiple groups, one-way ANOVA was used. For non-normally distributed data, the Mann-Whitney test was employed. U The test can be performed using either the Kaplan-Meier test or the Kruskal-Wallis test. Kaplan-Meier curves represent survival rates, and the Log-Rank test compares survival rates between groups. P <0.05 is statistically significant.

[0043] Example 1: Animal Experiment 1. Mouse model construction 1.1 Animal preparation and endocrine environment control 1) Ovariectomy: Eight-week-old wild-type C57BL / 6J female mice were purchased from Shandong Pengyue Experimental Animal Technology Co., Ltd. After one week of acclimatization, bilateral ovariectomy (OVX) was performed. The control group underwent sham surgery, which involved removing adipose tissue around the ovaries that was equivalent in volume to the ovaries.

[0044] 2) Estrogenic cycle detection: Starting from the 5th day post-surgery, vaginal cell smears were prepared from all mice at a fixed time each morning. Vaginal exfoliated cells from the Sham group mice were stained with H&E and examined microscopically. The estrous cycle stage was determined based on the relative proportions of leukocytes, nucleated epithelial cells, and keratinocytes. Mice in the interestrus phase were selected for subsequent experiments. Since the OVX group mice were in the interestrus phase for an extended period, their vaginal smears were only used to maintain operational consistency and did not require cycle determination.

[0045] 3) Estrogen supplementation: During the estrous cycle detection, mice in the OVX group were given estradiol valerate 0.15 mg / kg / day by gavage, while mice in the Sham group were given an equal amount of control solvent (Vehicle) to ensure that the estrogen levels in the OVX group mice and the Sham group mice in the estrus period were the same.

[0046] 1.2 Construction of sepsis ARDS model and FSH intervention 1) Model construction: Sham group mice in the interestrus period and OVX group mice of the same batch and the same number were injected intraperitoneally with LPS 10 mg / kg to construct a sepsis and sepsis-induced acute lung injury model (sepsis ADRS model). The control group was injected with an equal amount of control solvent (Vehicle).

[0047] 2) FSH intervention: One hour after model establishment, 30 IU / kg FSH or an equal volume of control solvent (Vehicle) was injected intraperitoneally. Based on OVX surgery, sepsis model establishment, and FSH intervention, the patients were divided into the following 6 groups:

[0048] 2. Survival Curve Plotting After the sepsis model was established, the time of death of mice in each group was observed and recorded, and Kaplan-Meier curves were plotted to represent the survival rate.

[0049] 3. H&E staining and lung injury pathology scoring Twenty-four hours after establishing the sepsis model, the lower lobe of the right lung of each group of mice was harvested, fixed in 4% paraformaldehyde for 24 hours, embedded in paraffin, sectioned, and stained with H&E. Pathological changes in the lung tissue were observed under a light microscope, including alveolar septal thickening, inflammatory cell infiltration, and alveolar hemorrhage. Semi-quantitative analysis was performed using the Smith scoring system. Six high-power fields (400×) were randomly selected from each section, and scores were assigned according to the severity of the lesions (0–4 points). The average score was used as the pathological damage score for that sample.

[0050] The pathological manifestations of a lung injury score of 4 include: significant thickening and congestion of the alveolar walls, a reduced number of alveolar cavities, some cavities occupied by serous fluid, a further increase in nucleated cells on the alveolar walls, focal aggregation of local inflammatory cells, and focal hemorrhage. A lung injury score of 3 also includes: significant thickening and congestion of the alveolar walls, a reduced number of alveoli, some cavities occupied by serous fluid, and a further increase in nucleated cells on the alveolar walls. A lung injury score of 2 includes: marked thickening and congestion of the alveolar walls, a significant increase in nucleated cells on the walls, serous fluid in the alveolar cavities, and a small amount of leaked red blood cells. A lung injury score of 1 indicates a mild lesion, slight thickening of the alveolar walls, a slight increase in red blood cells and inflammatory cells on the walls, and a small amount of serous exudate in the alveolar cavities. A lung injury score of 0 indicates normal lung tissue structure and no obvious histopathological damage.

[0051] 4. Detection of wet-to-dry density of lung tissue The upper lobe of the left lung of each group of mice was taken, the surface moisture was absorbed with filter paper and the wet weight was measured. The lung tissue was dried in a 60℃ oven for 48 h until constant weight was achieved and the dry weight was measured. The wet-to-dry weight ratio of lung tissue was calculated to quantify the degree of pulmonary edema.

[0052] 5. Experimental Results A mouse model of sepsis was established by intraperitoneal injection of LPS (corresponding to groups 2, 4, and 6). Figure 1 The results showed that during the 48-hour observation period, the survival rate of mice in the OVX+FSH group was significantly higher than that in the Sham group, and the difference was statistically significant. P <0.05), indicating that FSH can improve the prognosis of sepsis.

[0053] A mouse model of sepsis was established by intraperitoneal injection of LPS 24 h later. Figure 2 The results showed that the lung tissue of mice in the Sham group was significantly edematous and had multiple superficial hemorrhages; compared with the Sham group, the OVX+FSH group had reduced lung tissue edema and fewer superficial hemorrhages. Figure 3 The results showed that, compared with the Sham group, the wet-to-dry weight ratio of lung tissue was significantly lower in the OVX+FSH group. P The result was <0.01, indicating that FSH can improve vascular leakage. Figure 4 The results showed that alveolar structure destruction and inflammatory cell infiltration were observed in lung tissue sections of mice in the Sham group, while the degree of alveolar structure destruction and inflammatory cell infiltration was milder in lung tissue sections of the OVX+FSH group. Figure 5The results showed that after intraperitoneal injection of LPS in mice, the mean score was 3.5 in the Sham group, 3.4 in the OVX group, and 2.6 in the OVX+FSH group; the results indicated that the pathological score of lung injury was significantly lower in the OVX+FSH group compared with that in the Sham group. P <0.001). These results indicate that FSH can improve acute lung injury in septic mice.

[0054] Example 2 Cell Experiment 1. Cell Culture The immortalized human PMVECs were purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd., and cultured using a special culture medium. When the confluence density was about 95%, they were passaged, inoculated at a ratio of 1:3, and passaged every 3 days.

[0055] 2. Western blot assay 1) Detection of FSHR expression: Proteins were extracted from human PMVECs and human ovarian granulosa cell line (KGN), with human KGN as a positive control. The concentration was detected by bicinchoninic acid assay (BCA). After denaturation at 37℃ for 30 min, the protein was loaded onto the membrane. After constant voltage electrophoresis, the protein was transferred to a polyvinylidene fluoride (PVDF) membrane. Non-specific binding sites on the PVDF membrane were blocked with skim milk powder. The membrane was then incubated with FSHR antibody overnight, followed by secondary antibody incubation and imaging the next day.

[0056] 2) Detection of ICAM1 expression: Human PMVECs were seeded in 12-well plates. When the cell density reached 90%, the cells were treated with 10 μg / ml LPS or Vehicle for 24 h, and 10 ng / ml FSH or Vehicle was added simultaneously. Protein was extracted and the concentration was detected using BCA. After denaturation at 100℃ for 10 min, the protein was loaded onto a PVDF membrane. After constant voltage electrophoresis, the protein was transferred to a PVDF membrane, and non-specific binding sites were blocked with skim milk powder. The membrane was then incubated overnight with ICAM1 antibody, followed by secondary antibody incubation and imaging the next day.

[0057] 3. Immunofluorescence staining Human PMVECs were seeded onto cell slides. When the cell density reached 90%, the cells were treated with 10 μg / ml LPS or Vehicle for 24 h, with 10 ng / ml FSH or Vehicle added simultaneously. The cell slides were then fixed with 4% paraformaldehyde at room temperature for 15 min, and incubated with blocking buffer containing 25% donkey serum at room temperature for 1 h to block non-specific binding sites. The cells were then incubated overnight with VE-cadherin antibody, followed by secondary antibody incubation the next day and Hoechst nuclear staining. Images were acquired using a laser confocal microscope.

[0058] 4. RT-qPCR Human PMVECs were treated using the same method as immunofluorescence staining. After 24 h of LPS and FSH treatment, the reaction system was prepared according to the reverse transcription kit instructions. Genomic DNA was first removed, followed by reverse transcription to obtain cDNA. An RT-qPCR system was then prepared according to the kit instructions, and amplification was performed for 40 cycles at 95℃ for 5 s and 60℃ for 30 s. Glyceraldehyde-3-phosphate dehydrogenase (GDL) was used as the amplifying agent. GAPDH () is an internal parameter, used for calculation IL1B and IL6 The relative expression level of genes.

[0059] 5. Experimental Results Figure 6 The results showed that Western blot analysis confirmed the expression of FSHR in PMVECs, which appeared as a specific band. Figure 7 The results showed that after LPS treatment of human lung microvascular endothelial cells (PMVECs) for 24 h, immunofluorescence staining revealed multiple interruptions in intercellular connections in the control group (Vehicle) (as indicated by arrows); compared with the control group, the continuity of intercellular connections in the FSH-treated group was significantly improved and the interruptions were reduced. Figure 8 The results showed that, compared with the control group, FSH treatment significantly reduced the expression levels of inflammatory genes in PMVECs; among them, Figure 8 A displays the FSH group. IL1B Expression levels decreased significantly ( P <0.05), Figure 8 B displays the FSH group. IL6 The expression level was also significantly reduced. P <0.01). Figure 9The results showed that Western blot analysis indicated that the expression of intercellular adhesion molecule 1 (ICAM1), a marker of endothelial activation, was increased in the control group PMVECs; compared with the control group, the expression level of ICAM1 was significantly decreased in the FSH group.

[0060] This indicates that the control group showed elevated levels of ICAM1, a marker of endothelial activation, multiple interruptions in intercellular connections, and increased expression of inflammatory genes, suggesting damage to the pulmonary microvascular endothelial barrier. In contrast, the FSH group showed decreased ICAM1 levels, improved intercellular connectivity, and significantly reduced expression of inflammatory genes, indicating that FSH can inhibit PMVEC activation and thus stabilize the pulmonary microvascular endothelial barrier.

[0061] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The use of follicle-stimulating hormone in the preparation of medicaments for the prevention and / or treatment of acute lung injury caused by sepsis in subjects.

2. The application according to claim 1, characterized in that, The acute lung injury caused by sepsis is an acute complication of sepsis, which is caused by bacterial infection.

3. The application according to claim 1, characterized in that, The drug can be introduced into the body, such as into muscles, intradermal, subcutaneous, venous, or mucosal tissues, through injection, oral administration, spraying, penetration, absorption, or physical or chemical means; or it can be introduced into the body after being mixed with or encapsulated by other substances.

4. The application according to claim 1, characterized in that, Based on follicle-stimulating hormone (FSH), the single dose of the drug is 15-45 IU / kg.

5. The application according to claim 4, characterized in that, The drug is prepared in the form of dosage units, configured as an independent dose containing 900-2700 IU of follicle-stimulating hormone for single use, and configured as a dosage unit in at least one independent dose form.

6. The application according to claim 4, characterized in that, The drug is a unit dosage form of a solid or liquid preparation; the dosage form includes capsules, pills, tablets, powders, granules, lozenges, lozenges, melts, powders, solutions, suspensions, or emulsions.

7. The application according to claim 1, characterized in that, The drug is directly targeted to the lungs of the subject and taken up by the subject's lung tissue.

8. The application according to claim 1, characterized in that, The drug is a drug that improves vascular endothelial cell function under sepsis conditions or improves the pathological condition of acute lung injury in sepsis.

9. The application according to claim 8, characterized in that, The drugs that improve vascular endothelial cell function under sepsis conditions are those that improve endothelial cell barrier function, reduce monocyte adhesion, or lower the level of cell inflammation; the drugs that improve the pathological condition of acute lung injury in sepsis include those that improve the pathological structure of lung tissue, reduce pulmonary edema and tissue protein exudation, or improve the pulmonary vascular barrier capacity.

10. The application according to claim 9, characterized in that, The gene for inflammation is IL1B , IL6 .

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