Lung sympathetic nerve local photogenetic activation system and method for acute respiratory distress syndrome

Abstract

The invention discloses a lung sympathetic nerve local photogenetic activation system and method for acute respiratory distress syndrome. The system comprises a genetic expression injection module for injecting a drug for expressing photosensitive protein; the drug administration positioning module is used for injecting up-conversion nano particles UCNPs to a region adjacent to a sympathetic chain beside a thoracic vertebra T2-T5; and the excitation source irradiation module is used for irradiating the lung by adopting a 980 nm NIR laser / LED excitation light source and converting NIR into blue light through UCNPs so as to activate and express photosensitive protein. The system can realize a'reversible and programmable 'sympathetic nerve accurate activation means on the local part of the lung, selectively activates the lung sympathetic nerve endings / postpartum neurons and induces local NE release under the condition that an optical fiber does not need to be implanted, and reduces ARDS under the condition that the side effect of NE on the whole body system is reduced.

Description

Technical Field

[0001] This invention relates to the field of non-invasive neuromodulation and treatment of severe lung diseases, and in particular to a local optogenetic activation system and method for pulmonary sympathetic nerves in acute respiratory distress syndrome. Background Technology

[0002] Acute respiratory distress syndrome (ARDS) is a pathological condition characterized by disruption of the alveolar-capillary barrier and inflammatory imbalance.

[0003] Treatment for ARDS primarily focuses on supportive therapy, but targeted drugs are lacking. Besides supportive therapy, treatments include systemic administration of β2-receptor agonists and vagal / central sympathetic modulation. However, the efficacy of systemic administration of β2-receptor agonists is limited in large clinical studies. While vagal / central sympathetic modulation can affect inflammation, a Nature article indicates that activating DBH-positive neurons (sympathetic neurons) in the brain can alleviate inflammatory diseases such as colitis. This suggests that traditional optogenetics has limitations, including the need for visible light fiber optic implantation, significant invasiveness, and insufficient spatial selectivity.

[0004] In summary, among the current treatment options, (1) central C1 neuron activation is used to suppress inflammation through the spleen-adrenal pathway, but the overall sympathetic output of the central nervous system will mobilize the endocrine arm (adrenaline), and repeated stimulation is prone to response decay, resulting in unstable efficacy; (2) nebulized norepinephrine (NE) or systemic β-receptor agonists are used for intervention, but systemic NE / β2 receptor agonists are difficult to maintain an effective and safe concentration gradient in the lungs and there is a risk of adverse cardiovascular reactions; (3) deep optogenetics based on UCNP is used for non-invasive regulation of the central or peripheral nervous system, but the safety of UCNP particles in the human body still needs further verification.

[0005] Given the aforementioned traditional technical problems, current treatments for ARDS lack a precise method for sympathetic nerve activation that is "local, reversible, and programmable" in the lungs. Summary of the Invention

[0006] To overcome the shortcomings of the prior art, one of the objectives of this invention is to provide a local optogenetic activation system for the pulmonary sympathetic nerves in acute respiratory distress syndrome, so as to solve the above-mentioned traditional technical problems. Through this system, a precise means of sympathetic nerve activation that is "local, reversible, and programmable" in the lungs can be achieved.

[0007] The second objective of this invention is to provide a method for local optogenetic activation of pulmonary sympathetic nerves for acute respiratory distress syndrome (ARDS). This method selectively activates pulmonary sympathetic nerve endings / postganglionic neurons without the need for fiber optic implantation, inducing local release of norepinephrine (NE), thereby alleviating ARDS by reducing the systemic side effects of NE.

[0008] One of the objectives of this invention is achieved through the following technical solution: A local optogenetic activation system for pulmonary sympathetic nerves in patients with acute respiratory distress syndrome, comprising: The genetic expression injection module is used to inject drugs that express light-sensitive proteins, delivering the drugs to target neurons to achieve light-controlled activation properties in noradrenergic neurons. The drug delivery positioning module is used to inject upconversion nanoparticles (UCNPs) into the sympathetic chain adjacent to the T2–T5 thoracic vertebrae. The excitation source irradiation module is used to irradiate the lungs with an excitation source of 980 nm NIR (near-infrared) laser / LED. The NIR is converted into blue light by UCNPs to activate the expression of photosensitive proteins.

[0009] Furthermore, the local optogenetic activation system of the pulmonary sympathetic nervous system also includes: The monitoring module is used to monitor lung leakage, inflammatory markers, and respiratory mechanics indicators. The adjustment module is used to adjust the parameters of the excitation light source based on the lung indicators provided by the monitoring module. This allows for adaptive adjustment of the total dose of the excitation light source, the drug expressing the photosensitive protein, and the upconversion nanoparticles (UCNPs), thereby inducing local NE (norepinephrine) release. Preferably, the relationship between the frequency (1-20Hz), pulse width, and total dose of the excitation light source is adjusted to achieve the desired effect.

[0010] Furthermore, the expressed photosensitive protein is one of ChR2 or a functionally equivalent variant, DBH-Cre×AAV-DIO-ChR2, a selective photosensitive / magnetic channel, or a chemically targeted genetic drug.

[0011] Among them, functionally equivalent variants are one or more of hChR2(H134R), ChrimsonR, and C1V1, which can be activated by blue light (such as 473 nm), triggering cation influx, depolarizing the cell membrane, and thus rapidly activating neurons.

[0012] When applied to animal models, the light-sensitive protein is expressed as DBH-Cre×AAV-DIO-ChR2. DBH-Cre×AAV-DIO-ChR2 consists of two parts: DBH-Cre and AAV-DIO-ChR2. DBH-Cre is a transgenic mouse strain that expresses Cre recombinase under the promoter control of the dopamine β-hydroxylase (DBH) gene. Since DBH is a marker enzyme of norepinephrine neurons, Cre recombinase is specifically expressed in these neurons. AAV-DIO-ChR2 is a recombinant adeno-associated virus (AAV) vector. The ChR2 (channel rhodopsin-2) gene it carries is surrounded by two LoxP sites (DIO, i.e., double inverted open reading frames), ensuring that the ChR2 gene is expressed and translated into protein only in the presence of Cre recombinase.

[0013] When applied clinically, the expression of photosensitive proteins can be selective drug-sensitive / magnetic channels or chemogenetic targeted drugs, drug-sensitive / magnetic ion channels that have a highly selective response to specific small molecules or magnetic field stimulation (such as drug-dependent ChR2 variants, magnetic TRPV1 / TRPC channels, etc.), and targeted receptors based on chemogenetic tools such as DREADDs, to achieve reversible and precise regulation of specific neuronal populations.

[0014] Furthermore, the upconversion nanoparticles UCNPs are NaYF4:30%Yb, 0.5%Tm@NaYF4:75%Lu, and their surface is densely coated. Layer, in which, The layer thickness is preferably 2.5-3.5 nm. The thickness of the layer is 3 nm.

[0015] UCNPs have a core-shell structure, with the core being a NaYF4 matrix doped. and The outer shell is The co-doped NaYF4 layer is coated with a dense layer of silicon dioxide on its outermost layer. The protective layer, within the core layer, accounts for 30%. As a sensitizer, it efficiently absorbs near-infrared light (e.g., 980 nm) and transfers energy to the activator. 0.5% As an activator, it emits upconversion fluorescence (such as blue or near-infrared light) after energy transfer; within the shell, 75%... Co-doped NaYF4 shells can effectively suppress nonradiative relaxation, improve luminescence efficiency, and reduce the quenching effect of surface defects on luminescence. In the synthesis of core-shell structures, solvothermal or thermal decomposition methods are usually used to grow the core and shell layers stepwise. In the coating layer, dense The layer, formed via a sol-gel method, enhances the water solubility, biocompatibility, and chemical stability of nanoparticles, while providing active sites for surface functionalization (such as linking target molecules). It is typically formed on the nanoparticle surface through the hydrolysis of tetraethyl orthosilicate (TEOS) under alkaline conditions. layer.

[0016] Furthermore, upconversion nanoparticles (UCNPs) grafted with fluorescent tracer structures, such as Cy7, can be used to track the distribution of UCNPs in vivo.

[0017] Furthermore, the power of the excitation source is 1-3mW (e.g., 1mW, 2mW, 3mW, etc.), and this power range is adjustable. This power range can be adjusted by programming the duty cycle and frequency. Preferably, the power of the excitation source is 3mW.

[0018] The second objective of this invention is achieved by the following technical solution: A method for local optogenetic activation of pulmonary sympathetic nerves for acute respiratory distress syndrome includes the following steps: S1: Injecting a drug that expresses a photosensitive protein to deliver the drug to the target neuron; S2: Inject upconversion nanoparticles (UCNPs) into the sympathetic chain adjacent to the T2–T5 thoracic vertebrae; S3: The upconversion nanoparticles UCNPs are stably retained in the lungs. The lungs are irradiated with a 980nm NIR laser / LED excitation source according to the preset NIR parameters. The NIR is converted into blue light (~475 nm) by UCNPs to activate the expression of photosensitive proteins. S4: Monitoring and closed-loop regulation to achieve dynamic adjustment of NIR photostimulation parameters of the excitation source.

[0019] Further, in step S1, the expressed photosensitive protein is one of ChR2 or a functionally equivalent variant, DBH-Cre×AAV-DIO-ChR2, a selective photosensitive / magnetic channel, or a chemically targeted genetic drug.

[0020] Furthermore, in step S2, the upconversion nanoparticles UCNPs are NaYF4:30%Yb, 0.5%Tm@NaYF4:75%Lu, and their surface is densely coated. Layer, in which, The layer thickness is preferably 2.5-3.5 nm. The thickness of the layer is 3nm.

[0021] Furthermore, upconversion nanoparticles (UCNPs) grafted with fluorescent tracer structures, such as Cy7, can be used to track the distribution of UCNPs in vivo.

[0022] Furthermore, in step S3, the waiting time t ≥ 24 h is used in the step of stabilizing the upconversion nanoparticles UCNPs in the lungs, and the preset NIR parameters are 1-3 mW, 3-5 s / time, once every 2-4 h; preferably, the preset NIR parameters are 3 mW, 5 s / time, once every 4 h.

[0023] Further, in step S4, adjusting the NIR light stimulation parameters of the excitation source involves adjusting the frequency and pulse width of the NIR, wherein the frequency is 1-20Hz.

[0024] Furthermore, when the virus used to induce the expression of the photosensitive protein is At that time, the working titer of the drug expressing the photosensitive protein was 1×10¹² vg / mL, and the drug was administered by tracheal injection with an injection volume of 40 μL per animal. Among them, the upconversion nanoparticles used for in vivo optogenetic activation are UCNPs (and their Cy7-labeled form UCNPs-Cy7) were coated with upconversion nanoparticles at a concentration of 8 mg / mL, and a total volume of 25 μL was injected at both ends of the paravertebral sympathetic chain region in the T2–T5 segment.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. The local optogenetic activation system for pulmonary sympathetic nerves of the present invention is composed of modules such as a genetic expression injection module, a drug delivery positioning module, and an excitation source irradiation module, which can realize a precise activation method for sympathetic nerves that is "local, reversible, and programmable" in the lungs.

[0026] 2. The local optogenetic activation method of the pulmonary sympathetic nerves of the present invention selectively activates the pulmonary sympathetic nerve endings / postganglionic neurons without the need for optical fiber implantation, inducing local NE release, thereby reducing ARDS while minimizing the systemic side effects of NE. Attached Figure Description

[0027] Figure 1 The diagram shows the conversion luminescence mechanism and characterization of upconversion nanoparticles (UCNPs) in Example 1. Figure 1 A is a schematic diagram of the upconversion luminescence mechanism of UCNPs. Figure 1 B is a TEM image of UCNPs, scale bar: 50 nm; Figure 1 C is TEM image, scale bar: 50 nm; Figure 1 D is Upconversion emission spectrum and corresponding photographs under 980 nm excitation; Figure 1 E is the elemental distribution analysis diagram of UCNPs, scale bar: 20 nm; Figure 2 This is a schematic diagram of the mouse experiment in Example 4; Figure 3 This is a representative image of lung tissue from the EBA exudation experiment in mice in Example 4; Figure 4 The graph shows the quantitative analysis of EBA in mice from Example 4 (n=3). Figure 5 The graph shows the plasma TNF-α levels in mice from Example 4 (n=8). Figure 6 The graph shows the IL-6 levels in the plasma of mice in Example 4 (n=8). Figure 7 The graph shows the total protein concentration in the BALF of mice in Example 4 (n=5). Figure 8 The total lung injury score (n=5) of mice in Example 4 was calculated based on the histological features of H&E stained sections. Figure 9 Representative image of H&E stained lung tissue section from mice in Example 4, scale bar: 100 μm; Figure 10 This is a diagram illustrating the acute toxicological safety of mice in Example 4. Figure 10 A is a flowchart of an animal experiment on acute lung injury (ALI) in mice; Figure 10 B is a dynamic curve showing the change in mouse body weight over time; Figure 10 C is a statistical bar chart of lung injury scores in mice; Figure 10 D is a statistical bar chart of AST levels in mice; Figure 11 This is a representative anatomical diagram of the mouse paravertebral sympathetic trunk from Example 5. Figure 12 This is a quantitative analysis chart of norepinephrine (NE) concentration in bronchoalveolar lavage fluid (BALF) of Example 5 (n=8). Figure 13 The whole-cell patch-clamp recordings of Example 5 show the action potentials induced by light stimulation at 1 Hz, 10 Hz, and 20 Hz (pulse width 1 s, interval 1 s, laser power 3 mW). Detailed Implementation

[0028] The present invention will now be further described in conjunction with specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments. In the following embodiments, all materials are purchased externally and will not be described in detail here.

[0029] Example 1: Preparation of Upconversion Nanoparticles (UCNPs): The upconversion nanoparticles (UCNPs) were prepared first using a solvothermal method to create NaYF4:30%Yb,0.5%Tm@NaYF4:75%Lu core-shell structured nanocrystals. Specifically, 1 mmol of rare earth chlorides (YCl3, YbCl3, TmCl3, or LuCl3 in a molar ratio of NaYF4:30%Yb,0.5%Tm as the core and NaYF4:75%Lu as the shell) were added to a 100 mL three-necked flask. After removing the water, 15 mL of 1-octadecene (ODE) and 6 mL of oleic acid (OA) were added. The mixture was stirred at 156 °C for at least 20 min to form a rare earth-oleic acid complex. After the system was cooled to below 60 °C, a methanol solution containing 4 mmol NH4F and 2.5 mmol NaOH was added. After thorough stirring, the methanol was evaporated, and the mixture was purged with argon gas at least three times. The temperature was then raised to 300 °C and held for 50 min to obtain NaYF4:30%Yb,0.5%Tm core UCNPs. The product was precipitated with acetone and washed three times by centrifugation in a cyclohexane / acetone system. It was then redispersed in 20 mL of cyclohexane for later use. Subsequently, using these core UCNPs as seed crystals, the above steps of ODE / OA complexation, NH4F / NaOH introduction, and high-temperature reaction at 300 °C were repeated, with the high-temperature reaction time extended to 1.5 h. A NaYF4:75%Lu shell layer was epitaxially grown on the core surface to obtain NaYF4:30%Yb,0.5%Tm@NaYF4:75%Lu core-shell UCNPs.

[0030] Then, a sol-gel method was used to construct on its surface. Coating layer: Add 4 mL of cyclohexane dispersion containing UCNPs to 21 mL of cyclohexane, then add 1.5 mL of Igepal CO-520, sonicate for 30 s, and stir at 750 rpm for 5 min to form a stable reverse micelle system. Add 160 μL of 33% ammonia solution dropwise and continue stirring for 30 min. Then slowly add 80 μL of TEOS and react at room temperature for at least 16 h. In-situ hydrolysis and condensation on the particle surface forms a dense shell. The reaction solution is dispensed into 50 mL centrifuge tubes, and ethanol and acetone are added sequentially. After multiple rounds of vortexing, sonication, and washing by centrifugation at 12,000 rpm for 20 min, the precipitate is resuspended sequentially in an ethanol / water mixture and finally in deionized water. After sonication and homogenization, it is stored in glass bottles to obtain a product with good dispersibility and biocompatibility. Coated upconversion nanoparticles .

[0031] UCNPs composition: NaYF4:30%Yb,0.5%Tm@NaYF4:75%Lu.

[0032] The generated upconversion nanoparticles UCNPs, such as Figure 1 As shown, where, Figure 1 A is a schematic diagram of the upconversion luminescence mechanism of UCNPs. Figure 1 B is a TEM image of UCNPs, scale bar: 50 nm; Figure 1 C TEM image, scale bar: 50 nm; Figure 1 D is Upconversion emission spectrum and corresponding photographs under 980 nm excitation; Figure 1 E is the elemental distribution analysis diagram of UCNPs, scale bar: 20 nm.

[0033] from Figure 1 B and Figure 1 As can be seen from C, this Example 1 confirms the successful synthesis of UCNPs coated with a silica shell.

[0034] Example 2: Preparation of upconversion nanoparticles (UCNPs) grafted with fluorescent tracer structures: The upconversion nanoparticles (UCNPs) obtained in Example 1 were grafted with Cy7 via an esterification chemical reaction. Details are as follows: First, regarding UCNPs@SiO2 Aminoation modification: 20 mL Aqueous suspension (1 mg / mL) was mixed with 20 mL of anhydrous ethanol. Under magnetic stirring, 200 μL of 33% ammonia solution was added and pre-stirred for 30 min. Then, 20 μL of APTES was added dropwise, and the reaction was continued at room temperature with stirring for 24 h, allowing APTES to condense on the silica shell surface to form an amino-rich organosilicon layer. After the reaction, unreacted APTES and small molecule impurities were removed by multiple vortexing, sonication, and centrifugation with acetone / ethanol. Finally, the precipitate was resuspended in ethanol to obtain... Then, 10 mL The solution was changed to PBS buffer (1 mg / mL), and an equal volume of CY7-NHS ester solution (1 mg / mL) was added under stirring. The reaction was carried out at room temperature for 2–4 h or overnight at 4°C in the dark. Grafting of CY7 dye was achieved through amide bond coupling between the NHS ester and the surface primary amine. After the reaction was complete, the free CY7-NHS was removed by repeated centrifugation and washing with PBS or deionized water, finally obtaining surface covalently modified CY7. (Recorded as UCNPs-Cy7), and stored separately away from light for later use.

[0035] Implement a local optogenetic activation system for pulmonary sympathetic nerves in acute respiratory distress syndrome, comprising: a genetic expression injection module for injecting ChR2 to deliver ChR2 to target neurons to achieve photocontrolled activation in noradrenergic neurons; wherein, in the selection of ChR2, DBH-Cre×AAV-DIO-ChR2 can be used in animal models, and can be replaced by selective drug photosensitive / magnetic channels or chemical genetic targeting in clinical practice; A drug delivery positioning module is used to inject upconversion nanoparticles (UCNPs) obtained in Example 1 into the sympathetic chain adjacent to the T2–T5 thoracic vertebrae. The excitation source irradiation module is used to irradiate the lungs with an excitation source of 980 nm NIR (near-infrared) laser / LED, and convert NIR into blue light through UCNPs to activate the expression of photosensitive proteins; The monitoring module is used to monitor lung leakage, inflammatory markers, and respiratory mechanics indicators. The adjustment module is used to adjust the parameters of the excitation light source based on the lung indicators provided by the monitoring module, so as to adjust the total dose of the excitation light source, the drug expressing photosensitive protein, and the drug of upconversion nanoparticles UCNPs, so as to induce local NE (norepinephrine) release. The specific adjustment method depends on the specific situation and will not be described in detail here.

[0036] Example 4 Mouse experiment: as Figure 2 The diagram shown illustrates a mouse experiment, administered via intratracheal injection. Retrograde transduction of sympathetic neurons was achieved in DBH-Cre mice. After 21 days, UCNPs were percutaneously injected near the paravertebral sympathetic trunk. ARDS was induced by intratracheal administration of LPS (5 mg / kg), followed by 980 nm near-infrared irradiation (3 mW, 5 seconds every 4 hours for 24 hours). Lung tissue and fluid were collected 3 hours later for evaluation.

[0037] Among them, the virus that induces the expression of photosensitive protein is At that time, the working titer of the drug expressing the photosensitive protein was 1×10¹² vg / mL, and the drug was administered by tracheal injection with an injection volume of 40 μL per animal. Upconversion nanoparticles for in vivo optogenetic activation are UCNPs (and their Cy7-labeled form UCNPs-Cy7) were coated with upconversion nanoparticles at a concentration of 8 mg / mL, and a total volume of 25 μL was injected at both ends of the paravertebral sympathetic chain region in the T2–T5 segment.

[0038] Evaluation content such as Figures 3-10 As shown, where, Figure 3 Representative images of lung tissue from mice in the EBA exudation experiment; Figure 4 Graphs showing quantitative analysis of EBA in mice (n=3); Figure 5 A graph showing the plasma TNF-α levels in mice (n=8); Figure 6 A graph showing the plasma IL-6 levels in mice (n=8); Figure 7 The graph shows the total protein concentration in the BALF of mice (n=5). Figure 8 Total lung injury score map calculated for mice based on histological features of H&E stained sections (n=5); Figure 9 Representative image of H&E stained lung tissue sections from mice, scale bar: 100 μm. Figure 10 The diagram shows the acute toxicological safety of mice. Figure 10 A is a flowchart of an animal experiment on acute lung injury (ALI) in mice. Through pretreatment, modeling, sacrifice and analysis, tissue samples such as blood, liver, and kidney are collected for subsequent indicator detection and pathological analysis. Figure 10 B is a dynamic curve of mouse body weight change over time, showing the effect of UCNPs intervention on the overall condition (body weight) of mice after LPS-induced ALI, reflecting the trend of the overall health level of mice over time. Figure 10 C is a statistical bar chart of lung injury scores in mice, comparing the degree of pathological damage to lung tissue under different intervention conditions, and quantifying the protective effect of UCNPs against LPS-induced lung injury. Figure 10 D is a statistical bar chart of AST levels in mice. AST is a sensitive indicator of liver damage. By comparing the serum AST levels in different groups through the bar chart, the intervention effect of UCNPs on LPS-induced liver inflammation and functional impairment can be evaluated.

[0039] As can be seen from the above, UCNPs-mediated optogenetic activation can alleviate LPS-induced ARDS in mice, specifically: 1) AAV is delivered to the lungs via nebulization, retrogradely transducing pulmonary sympathetic projection neurons; 2) In vitro / ex vivo segmental recordings show that 1-20 Hz light stimulation can induce action potentials and NE release; 3) In the LPS-induced ARDS model, the combination of "UCNPs+NIR+ChR2" significantly reduces EBA extravasation, BALF protein, plasma TNF-α / IL-6, and pathological scores; 4) Particles are locally enriched in the paravertebral sympathetic chain, with low accumulation in major organs; 5) Acute toxicological safety is good (no significant increase in AST / ALT / BUN).

[0040] Example 5 Monitoring and Closed-Loop Regulation Experiment: Based on Example 4, Example 5 uses patch-clamp technique to compare changes in sympathetic nerve excitation and norepinephrine (NE) release levels under different light stimulation parameters. Specific results are as follows: Figures 11-13 ,in, Figure 11 This is a representative anatomical diagram of the paravertebral sympathetic trunk in mice (shown by white arrows). Three weeks after intrapulmonary injection of AAV-DIO-ChR2 into DBH-Cre mice, viral expression was localized in this region. Subsequently, the sympathetic trunk was dissected under a stereomicroscope for electrophysiological and related functional experiments. Figure 12 A quantitative analysis graph of norepinephrine (NE) concentration in bronchoalveolar lavage fluid (BALF) (n=8). Figure 13 Whole-cell patch-clamp recordings show action potential maps induced by light stimulation at 1 Hz, 10 Hz, and 20 Hz (pulse width 1 s, interval 1 s, laser power 3 mW).

[0041] The results above show that continuous electrical activity peaks were only detected when UCNPs and NIR light were present simultaneously.

[0042] In summary, the method of the present invention has the following advantages: 1. Long-term retention of UCNPs in the paravertebral sympathetic chain of the T2–T5 thoracic vertebrae and upconversion triggered by NIR blue light achieve “in vivo–local–reversible” neural activation.

[0043] 2. Genetic or equivalent methods can be used to make the pulmonary sympathetic projection nerves sensitive to blue light, thereby inducing local NE release and exerting a barrier protection through the β2-AR–cAMP–PKA–VASP axis.

[0044] 3. Closed-loop optimization method for therapeutic photostimulation prescriptions (frequency / pulse width / total dose / interval pattern) for ARDS.

[0045] Compared with existing "central sympathetic activation / systemic NE or β agonists", this invention emphasizes "local, non-invasive, and programmable" activation of sympathetic terminals and barrier protection in the lungs. Its technical effect is to significantly reduce permeability and inflammation without introducing systemic side effects.

[0046] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A local optogenetic activation system for pulmonary sympathetic nerves in acute respiratory distress syndrome, characterized in that, include: The genetic expression injection module is used to inject drugs that express light-sensitive proteins, delivering the drugs to target neurons to achieve light-controlled activation properties in noradrenergic neurons. The drug delivery positioning module is used to inject upconversion nanoparticles (UCNPs) into the sympathetic chain adjacent to the T2–T5 thoracic vertebrae. The excitation source irradiation module is used to irradiate the lungs with an excitation source of 980 nm NIR laser / LED, and converts NIR into blue light through UCNPs to activate the expression of photosensitive proteins.

2. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 1, characterized in that, The expressed photosensitive protein is one of ChR2 or a functionally equivalent variant, DBH-Cre×AAV-DIO-ChR2, a selective photosensitive / magnetic channel, or a chemically targeted genetic drug.

3. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 1, characterized in that, The upconversion nanoparticles (UCNPs) are NaYF4:30%Yb, 0.5%Tm@NaYF4:75%Lu, and their surface is densely coated. layer.

4. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 3, characterized in that, The The thickness of the layer is 2.5-3.5 nm.

5. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 3, characterized in that, The upconversion nanoparticles UCNPs are grafted with fluorescent tracer structures.

6. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 5, characterized in that, The fluorescent tracer structure is Cy7.

7. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 1, characterized in that, The power of the excitation light source is 1-3mW.

8. The pulmonary sympathetic local optogenetic activation system for acute respiratory distress syndrome according to claim 1, characterized in that, The pulmonary sympathetic local optogenetic activation system also includes: The monitoring module is used to monitor lung leakage, inflammatory markers, and respiratory mechanics indicators. The adjustment module is used to adjust the parameters of the excitation light source according to the lung indicators provided by the monitoring module, so that the total dose of the excitation light source, the drug expressing photosensitive protein, and the drug upconversion nanoparticles UCNPs can be adaptively adjusted to induce local NE release.

9. A method for local optogenetic activation of pulmonary sympathetic nerves for acute respiratory distress syndrome, characterized in that, Includes the following steps: S1: Injecting a drug that expresses a photosensitive protein to deliver the drug to the target neuron; S2: Inject upconversion nanoparticles (UCNPs) into the sympathetic chain adjacent to the T2–T5 thoracic vertebrae; S3: The upconversion nanoparticles UCNPs are stably retained in the lungs. The lungs are irradiated with a 980 nm NIR laser / LED excitation source according to the preset NIR parameters. The NIR is converted into blue light by the UCNPs to activate the expression of photosensitive proteins. S4: Monitoring and closed-loop regulation to achieve dynamic adjustment of NIR photostimulation parameters of the excitation source.

10. The method for local optogenetic activation of pulmonary sympathetic nerves for acute respiratory distress syndrome according to claim 9, characterized in that, In step S3, the preset NIR parameters are 1-3 mW, 3-5 s / time, once every 2-4 h.

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