Protective agent for lung injury caused by intestinal ischemia reperfusion
By chemically modifying triptycholin, a protective agent for lung cells was prepared, which solved the problem of lung damage caused by intestinal ischemia and reperfusion injury and achieved effective protection of lung cells.
Patent Information
- Application Number
- CN202411779234.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-05
- Publication Date
- 2025-03-18
AI Technical Summary
Intestinal ischemia and reperfusion injury often leads to lung injury, and its mechanism has not been fully clarified, resulting in increased treatment difficulty.
By chemically modifying triptyrene to improve its bioavailability and antioxidant ability, a protective agent for lung cells was prepared to reduce lung cell coking and oxidative stress damage.
This protective agent can effectively reduce the scorching of lung cells, prevent the cells from releasing inflammatory mediators, avoid further aggravating the inflammatory response and cell damage, thereby protecting the lungs.
Smart Images
Figure CN119632995A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of lung injury protective agents, in particular to a protective agent for lung injury caused by intestinal ischemia-reperfusion. Background Art
[0002] Intestinal ischemia-reperfusion injury is one of the common tissue and organ injuries. It plays an important role in the pathophysiological evolution of diseases such as intestinal obstruction, severe infection, shock, and surgeries such as extracorporeal circulation and small intestinal transplantation. Intestinal ischemia-reperfusion injury can not only induce local intestinal damage, but also induce multiple organ failure due to intestinal bacterial translocation and bacterial toxins, and eventually lead to death. It has the characteristics of high incidence and mortality, which seriously endangers human health and safety.
[0003] The lung is one of the most vulnerable organs after intestinal ischemia-reperfusion injury. The main clinical manifestations are acute lung injury and respiratory distress syndrome. Lung injury is more serious than intestinal injury and is the main cause of death in patients with intestinal ischemia-reperfusion injury. Therefore, acute lung injury induced by intestinal ischemia-reperfusion injury is an issue worthy of attention.
[0004] After a patient suffers from lung injury, they may suffer from long-term lung function damage, which seriously affects the patient's quality of life and survival rate. When treating intestinal ischemia-reperfusion injury, the protection of the lungs should be strengthened as soon as possible to prevent and treat the damage to the body caused by it and improve the patient's prognosis and quality of life. Preliminary research has been conducted on the mechanism of lung injury caused by intestinal ischemia-reperfusion injury, but the exact molecular mechanism of its occurrence and development has not yet been clarified. Therefore, in-depth research on the occurrence and development of acute lung injury caused by intestinal ischemia-reperfusion injury will help reveal the specific molecular mechanism of the occurrence and development of acute lung injury caused by intestinal ischemia-reperfusion injury, and provide new targets and measures for the treatment of acute lung injury caused by intestinal ischemia-reperfusion injury. It is of great significance to further improve the treatment level of acute lung injury induced by intestinal ischemia-reperfusion injury.
[0005] To this end, we provide a protective agent against intestinal ischemia-reperfusion-induced lung injury to solve the above-mentioned problems. Summary of the invention
[0006] The purpose of the present invention is to provide a protective agent for lung injury caused by intestinal ischemia-reperfusion, and to solve the problem of lung cell charring caused by intestinal ischemia-reperfusion injury by chemically modifying tripterygium wilfordii to improve its bioavailability and enhance its antioxidant effect in lung cells.
[0007] In order to solve the above technical problems, the present invention is achieved through the following technical solutions:
[0008] The present invention is a protective agent for lung injury caused by intestinal ischemia-reperfusion, comprising the following steps: Step a: Synthesis step: S1. Purification process of tripterygium wilfordii: Stir the obtained concentrated solution with silica gel to fully mix the silica gel and the concentrated solution, dry them into dispersed particles to obtain a granular raw material for standby use, place the silica gel in a dichloromethane-methanol solution for wet column loading to ensure that the silica gel is evenly distributed in the column, balance the separation column with a dichloromethane-methanol solution to remove impurities and bubbles in the column, and prepare to receive the granular raw material, after the granular raw material is loaded on the column, use an eluent for gradient elution, and add a small amount of anhydrous sodium sulfate to remove moisture in the eluent. During the elution process, according to the Select a suitable eluent ratio according to the polarity of celastrol to separate and purify celastrol, collect the eluent to obtain a crude celastrol solution, recover the crude celastrol solution under reduced pressure, remove the eluent, obtain pure celastrol, wash the pure celastrol with methanol or anhydrous ethanol, further remove impurities, and obtain celastrol monomer; S2. Chemical modification: dissolve the celastrol raw material in an appropriate solvent, dissolve the esterification agent in the same solvent, and slowly add it dropwise to the celastrol solution while stirring and mixing, carry out esterification reaction of celastrol by heating and stirring, and monitor by thin layer chromatography, high performance liquid chromatography and other methods. The reaction conditions are adjusted according to the reaction results. After the reaction is completed, the reaction solution is cooled to room temperature, solid impurities are removed by filtration to obtain a clarified reaction solution, and the reaction solution is extracted with an appropriate solvent to extract the target product. The extracts are combined and dried to remove water from the solvent. The extracted products are purified by column chromatography, recrystallization and other methods, and the purified products are collected and dried to obtain the target esterified derivatives. S3. Product identification and testing stage: The target esterified derivatives are structurally identified using methods such as nuclear magnetic resonance and infrared spectroscopy to confirm whether the chemical structure of the product meets expectations, and the target esterified derivatives are subjected to biological activity testing. Step b: Model establishment: S1. Preparation for the cultivation of human alveolar epithelial cells (A549 cells): Set the temperature of the cell culture incubator to provide a suitable environment for cell growth, take out the frozen A549 cells from liquid nitrogen, and quickly put them into a water bath to thaw. During this period, gently shake the cryotube to promote rapid thawing of the cells; S2. Cultivation of human alveolar epithelial cells (A549 cells): Add the thawed cell suspension to a centrifuge tube, discard the supernatant after centrifugation, and then add an appropriate amount of preheated culture medium to inoculate the resuspended cells into a culture bottle, add an appropriate amount of culture medium, and then place it in a cell culture incubator for cultivation; S3.Cytotoxicity test: using CCK-8 reagent detection method, cells with different concentrations of tripterygium wilfordii derivatives were inoculated into 96-well plates, incubated for a certain period of time after adding CCK-8 reagent, and then the absorbance value was measured, the cell viability was calculated according to the absorbance value, and the effect of different concentrations of tripterygium wilfordii derivatives on cell viability was measured, and the cell apoptosis rate after adding different concentrations of tripterygium wilfordii derivatives was detected by flow cytometry. After the cells were collected and processed, they were stained with fluorescently labeled annexin V and viability dyes (such as propidium iodide), and then analyzed by flow cytometry to obtain the cell apoptosis rate; Step c: Animal model: S1. Preoperative preparation of animal model: Select healthy mice with moderate weight, usually male mice, and the age and breed should be consistent. 1% sodium pentobarbital was injected intraperitoneally at a dose of 50 mg / kg into the mice. After anesthesia, use a razor to remove the abdominal hair and fix the mouse in a supine position on a warming pad to reduce contamination during the operation; S2. Surgical operation: vertically cut the skin and peritoneum along the midline of the mouse's abdomen, separate and clamp the superior mesenteric artery after opening the abdomen, release the non-invasive artery clamp after clamping for 45min-1h, perform reperfusion for 4h, suture the surgical incision with sterile sutures, and then disinfect the surgical area again, put the mouse back into the cage, give normal feeding and observation; S3. Drug administration and detection: Administer tripterygium wilfordii derivatives by oral or intravenous injection. 24h after administration, evaluate the degree of lung damage by lung tissue section and pathological examination, measure the levels of inflammatory factors (such as IL-6, TNF-α, etc.) in blood and lung tissue, and measure oxidative stress indicators (such as MDA, SOD, etc.) in lung tissue. .
[0009] The present invention is further configured that in the step a, the concentrated solution of tripterygium wilfordii and silica gel are stirred by a stirrer for 50-80 minutes, the granular raw material mixed with the solution is dried by a dryer, the eluent is composed of a dichloromethane-methanol solution and ammonia water, and the eluent is decompressed and recovered by a vacuum decompression recovery machine.
[0010] The present invention is further configured such that the solvent for dissolving the tripterygium wilfordii raw material in step a is ethanol, the heating temperature of the esterification reagent and the solution is 50-150°, which needs to be adjusted according to the reaction conditions, and the esterified derivative is dried by a dryer.
[0011] The present invention is further configured such that in step b, the culture temperature inside the cell culture box is set to 37°C, the carbon dioxide concentration is 5%, and the cell thawing water bath temperature is 37°C.
[0012] The present invention is further configured as follows: the centrifugal speed of the centrifuge tube in step b is 1000RPM, the centrifugation is 3-5min, the culture medium is a high-glucose DMEM culture medium containing 10% fetal bovine serum, and the culture medium needs to be preheated to 37° before use.
[0013] The present invention is further configured that, in step c, the mice are fasted for 12 hours before modeling, the mice should be allowed to drink water freely before surgery, the surgical area is disinfected with alcohol or iodine, and sterile gauze is covered on the thermal pad.
[0014] The present invention is further configured as follows: in step c, the length of the abdominal incision of the mouse is about 3-5 cm, the surgical area is covered with sterile gauze soaked in saline, the superior mesenteric artery is clamped with a microvascular clamp, the free intestine is wrapped with sterile gauze, and 37°C saline is dripped to maintain body temperature and moisture.
[0015] The present invention is further configured that, in step c, after the operation on the mice is completed, an appropriate amount of antibiotics is given to the mice, and the weight changes, survival rates and related indicators after intestinal ischemia-reperfusion injury of the mice are recorded.
[0016] The present invention has the following beneficial effects:
[0017] 1. The present invention is verified by establishing cell models and biological models, and can effectively reduce lung cell coking and protect the lungs from oxidative stress damage. When conducting biological model building, an intestinal ischemia-reperfusion injury model is established in mice using active modeling surgery to simulate clinical lung injury conditions. The tripterygium wilfordii derivatives are administered orally or intravenously to verify that they have the ability to inhibit lung cell coking, prevent cells from releasing histidine and a variety of inflammatory mediators, and avoid further aggravation of inflammatory reactions and cell damage.
[0018] 2. The present invention structurally transforms celastrol through chemical modification and successfully synthesizes a celastrol derivative with higher bioavailability, so that it can bind to TIPE2, avoid the shearing of GSDMD under the action of NLRP3, release GSDMD-NT, form holes in the cell membrane, lead to the release of cell contents, and effectively prevent it from inducing a strong inflammatory response that triggers cell pyroptosis. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for describing the embodiments are briefly introduced below.
[0020] Figure 1 The overall flow chart of a protective agent against intestinal ischemia-reperfusion-induced lung injury;
[0021] Figure 2 This is a HE staining validation diagram of a protective agent against lung injury caused by intestinal ischemia-reperfusion;
[0022] Figure 3 Schematic diagram of the establishment of an animal model of protective agents for lung injury caused by intestinal ischemia-reperfusion. DETAILED DESCRIPTION
[0023] The technical solutions in the embodiments of the present invention will be described below in conjunction with the drawings in the embodiments of the present invention. The described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.
[0024] Example 1
[0025] Reference Figure 1-Figure 3, which is the first embodiment of the present invention, and this embodiment provides a protective agent for lung injury caused by intestinal ischemia-reperfusion, including the following steps: Step a: Synthesis step: S1. Purification process of tripterygium wilfordii: Stir the obtained concentrated solution with silica gel to fully mix the silica gel and the concentrated solution, dry them into dispersed particles to obtain a granular raw material for use, place the silica gel in a dichloromethane-methanol solution for wet column loading to ensure that the silica gel is evenly distributed in the column, balance the separation column with a dichloromethane-methanol solution to remove impurities and bubbles in the column, and prepare to receive the granular raw material, after the granular raw material is loaded on the column, use an eluent for gradient elution, and add a small amount of anhydrous sodium sulfate to remove the eluent. The water in the deionizer, during the elution process, a suitable eluent ratio is selected according to the polarity of tripterygium wilfordii to separate and purify tripterygium wilfordii, the eluent is collected to obtain a crude solution of tripterygium wilfordii, the crude solution of tripterygium wilfordii is decompressed and recovered, the eluent is removed to obtain a pure product of tripterygium wilfordii, the pure product of tripterygium wilfordii is washed with methanol or anhydrous ethanol, and impurities are further removed to obtain a monomer of tripterygium wilfordii; in step a, the concentrated solution of tripterygium wilfordii and silica gel are stirred by a stirrer for 50-80 minutes, and the granular raw material mixed with the solution is dried by a dryer, the eluent washing is composed of a dichloromethane-methanol solution and aqueous ammonia, and the washing The deagent is recovered by vacuum decompression recovery machine; S2. Chemical modification: dissolve the raw material of tripterygium wilfordii in an appropriate solvent, dissolve the esterification agent in the same solvent, and slowly add it dropwise to the tripterygium wilfordii solution, stirring and mixing at the same time, esterifying the tripterygium wilfordii by heating and stirring, monitoring the progress of the reaction by thin layer chromatography, high performance liquid chromatography and other methods, adjusting the reaction conditions according to the reaction results, cooling the reaction solution to room temperature after the reaction, filtering to remove solid impurities, obtaining a clarified reaction solution, extracting the reaction solution with an appropriate solvent to extract the target product, combining the extracts, and drying to remove the impurities in the solvent. Water, purify the extracted product by column chromatography, recrystallization and other methods, collect the purified product, and dry it to obtain the target esterified derivative; the solvent for dissolving the raw material of tripterygium wilfordii in step a is ethanol, the heating temperature of the esterification reagent and the solution is 50-150°, which needs to be adjusted according to the reaction situation, and the esterified derivative is dried by a dryer; S3. Product identification and testing stage: use nuclear magnetic resonance, infrared spectroscopy and other methods to identify the structure of the target esterified derivative, confirm whether the chemical structure of the product meets the expectations, and conduct biological activity tests on the target esterified derivative to evaluate its activity; Step b: Model establishment: S1.Preparation for the cultivation of human alveolar epithelial cells (A549 cells): Set the temperature of the cell culture incubator to provide a suitable environment for cell growth, take out the frozen A549 cells from liquid nitrogen, and quickly put them into a water bath to thaw, and gently shake the cryotubes during the process to promote rapid thawing of the cells; in step b, the culture temperature inside the cell culture incubator is set to 37°C, the carbon dioxide concentration is 5%, and the cell thawing water bath temperature is 37°C; S2. Cultivation of human alveolar epithelial cells (A549 cells): Add the thawed cell suspension to a centrifuge tube, discard the supernatant after centrifugation, and then add an appropriate amount of preheated culture medium to inoculate the resuspended cells into a culture bottle, add an appropriate amount of culture medium, and then put the cell culture bottle into the culture bottle. Incubator for culturing; Step b: The centrifugal tube is centrifuged at a speed of 1000RPM for 3-5min, and the culture medium is a high-glucose DMEM culture medium containing 10% fetal bovine serum, which needs to be preheated to 37° before use; S3. Cytotoxicity test: Using CCK-8 reagent detection method, cells with different concentrations of tripterygium wilfordii derivatives are inoculated into 96-well plates, CCK-8 reagent is added and incubated for a certain period of time, and then the absorbance value is measured, and the cell viability is calculated according to the absorbance value, and the effect of different concentrations of tripterygium wilfordii derivatives on cell viability is measured, and the cell apoptosis rate after adding different concentrations of tripterygium wilfordii derivatives is detected by flow cytometry, and the cells are collected and processed, and then the membrane is labeled with fluorescent markers. The cells were stained with protein V and a viability dye (such as propidium iodide), and then analyzed by flow cytometry to obtain the cell apoptosis rate; Step c: Animal model: S1. Animal model preoperative preparation: Select healthy mice with moderate weight, usually male mice, and keep the same age and breed. 1% sodium pentobarbital was injected intraperitoneally at a dose of 50 mg / kg. After the mice were anesthetized, the abdominal hair was removed with a razor and the mice were fixed in a supine position on a warming pad to reduce contamination during the operation; Step c: The mice were fasted for 12 hours before modeling. Before the operation, the mice should be allowed to drink water freely. The surgical area was disinfected with alcohol or iodine, and sterile gauze was placed on the warming pad; S2. Surgical operation: The skin was cut vertically along the midline of the mouse's abdomen. Skin and peritoneum, etc., separate and clamp the superior mesenteric artery after opening the abdomen, release the non-invasive artery clamp after clamping for 45min-1h, perform reperfusion for 4h, suture the surgical incision with sterile sutures, disinfect the surgical area again, put the mice back into the cage, and give normal feeding and observation; Step c The length of the abdominal incision of the mouse is about 3-5cm, cover the surgical area with sterile gauze soaked in saline, clamp the superior mesenteric artery with a microvascular clamp, wrap the free intestine with sterile gauze, and drip 37℃ saline to maintain body temperature and moisture. Step c After the operation, the mouse is given an appropriate amount of antibiotics, and the weight change, survival rate and related indicators after intestinal ischemia-reperfusion injury of the mouse are recorded; S3.Administration test: The tripterygium wilfordii derivatives were administered orally or intravenously. 24 hours after administration, the lung tissue sections and pathological examinations were performed to evaluate the degree of lung damage, measure the levels of inflammatory factors (such as IL-6, TNF-α, etc.) in the blood and lung tissues, and measure the oxidative stress indicators (such as MDA, SOD, etc.) in the lung tissues.
[0026] Example 2
[0027] Reference Figure 1-Figure 3 , which is the second embodiment of the present invention, and this embodiment is based on the previous embodiment.
[0028] Amidation modification of tripterygium wilfordii
[0029] Weigh an appropriate amount of tripterygium wilfordii as a reaction substrate, dissolve tripterygium wilfordii and PyBOP in an appropriate amount of DMF solvent, place the reaction solution in an ice-water mixture to cool for a period of time to lower the reaction temperature and promote the dissolution of the reactants, slowly drip DIPEA into the reaction solution under stirring to neutralize the acid produced in the reaction and promote the formation of amide bonds, and after the reaction solution returns to room temperature, add glucosamine solution dropwise, and continue stirring for a period of time until the reaction is complete.
[0030] Post-reaction treatment
[0031] The reaction solution was extracted with dichloromethane and saturated brine to remove impurities, the organic layer was dried with anhydrous sodium sulfate to remove residual moisture, and reduced pressure rotary evaporation was performed at an appropriate temperature to remove the solvent and obtain a crude product, and the crude product was purified by silica gel column chromatography to obtain the target product, celastrol-glucosamine amide conjugate.
[0032] Example 3
[0033] Reference Figure 3 , which is the third embodiment of the present invention, and is based on the first two embodiments.
[0034] Comparative Animal Model Preparation
[0035] Healthy male SD rats were selected to ensure that the rats were adaptively raised before the experiment to reduce stress response. The rats were randomly divided into a sham operation group, a model group, and a drug-treated group. The number of each group was determined according to the experimental design. The sham operation group was used as a control, the model group was used to construct an intestinal ischemia-reperfusion injury model, and the drug-treated group was used to observe the effects of drugs on the model. The blood perfusion of the superior mesenteric artery was restored by clamping the superior mesenteric artery for 45 minutes, and the abdominal incision was sutured. The rats were given postoperative care, including maintaining a suitable environmental temperature and avoiding severe vibration and noise interference.
[0036] Comparative test results
[0037] Esterified tripterygium wilfordii and amidated tripterygium wilfordii were administered orally and intravenously. The rats in the drug-treated group were dissected and lung tissue sections were obtained 24 hours after administration. The protective effects of the derivatives were evaluated by the levels of inflammatory factors in tissue sections, blood and lung tissues, and oxidative stress indicators.
[0038] The preferred embodiments of the present invention disclosed above are only used to help explain the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that technicians in the relevant technical field can better understand and utilize the present invention.
Claims
1. A protective agent for lung injury caused by intestinal ischemia-reperfusion, characterized in that: The following steps are involved: Step a: Synthesis steps: S1. Purification process of tripterygium wilfordii: stirring the obtained concentrated solution with silica gel to fully mix the silica gel and the concentrated solution, drying into dispersed particles to obtain a granular raw material for standby use, placing the silica gel in a dichloromethane-methanol solution for wet column loading to ensure that the silica gel is evenly distributed in the column, balancing the separation column with a dichloromethane-methanol solution to remove impurities and bubbles in the column, and preparing to receive the granular raw material, after the granular raw material is loaded on the column, gradient elution is performed with an eluent, and a small amount of anhydrous sodium sulfate is added to remove the water in the eluent. During the elution process, a suitable eluent ratio is selected according to the polarity of tripterygium wilfordii to separate and purify tripterygium wilfordii, and the eluent is collected to obtain a crude solution of tripterygium wilfordii, the crude solution of tripterygium wilfordii is decompressed and recovered, and the eluent is removed to obtain a pure product of tripterygium wilfordii, and the pure product of tripterygium wilfordii is washed with methanol or anhydrous ethanol to further remove impurities to obtain a monomer of tripterygium wilfordii; S2. Chemical modification: dissolve the raw material of tripterygium wilfordii in an appropriate solvent, dissolve the esterification reagent in the same solvent, and slowly add it dropwise to the tripterygium wilfordii solution while stirring and mixing, and perform esterification reaction on tripterygium wilfordii by heating and stirring, and monitor the progress of the reaction by thin layer chromatography, high performance liquid chromatography and other methods, and adjust the reaction conditions according to the reaction results. After the reaction is completed, cool the reaction solution to room temperature, filter to remove solid impurities, and obtain a clarified reaction solution. Use an appropriate solvent to extract the reaction solution to extract the target product, combine the extracts, and dry them to remove water from the solvent. Purify the extracted product by column chromatography, recrystallization and other methods, collect the purified product, and dry it to obtain the target esterified derivative; S3. Product identification and testing stage: Use nuclear magnetic resonance, infrared spectroscopy and other methods to identify the structure of the target esterified derivatives, confirm whether the chemical structure of the product meets expectations, and conduct biological activity tests on the target esterified derivatives to evaluate their activity; Step b: Model establishment: S1. Preparation for the cultivation of human alveolar epithelial cells (A549 cells): Set the temperature of the cell culture incubator to provide a suitable environment for cell growth, take out the frozen A549 cells from liquid nitrogen, and quickly thaw them in a water bath. During this period, gently shake the cryotube to promote rapid thawing of the cells; S2. Cultivation of human alveolar epithelial cells (A549 cells): add the thawed cell suspension into a centrifuge tube, discard the supernatant after centrifugation, then add an appropriate amount of preheated culture medium, inoculate the resuspended cells into a culture bottle, add an appropriate amount of culture medium, and then place in a cell culture incubator for cultivation; S3. Cytotoxicity test: using CCK-8 reagent detection method, cells with different concentrations of tripterygium wilfordii derivatives were inoculated into 96-well plates, incubated for a certain period of time after adding CCK-8 reagent, and then the absorbance value was measured, and the cell viability was calculated according to the absorbance value, and the effects of different concentrations of tripterygium wilfordii derivatives on cell viability were measured, and the cell apoptosis rate after adding different concentrations of tripterygium wilfordii derivatives was detected by flow cytometry, and the cells were collected and processed, and then stained with fluorescently labeled annexin V and viability dyes (such as propidium iodide), and then analyzed by flow cytometry to obtain the cell apoptosis rate; Step c: Animal model: S1. Animal model preoperative preparation: Select healthy mice with moderate weight, usually male mice, of the same age and breed, and inject 1% sodium pentobarbital into the mouse's peritoneum at a dose of 50 mg / kg. After the mouse is anesthetized, use a razor to remove the abdominal hair and fix it in a supine position on a warming pad to reduce contamination during the operation; S2. Surgical operation: Cut the skin and peritoneum vertically along the midline of the mouse abdomen, separate and clamp the superior mesenteric artery after opening the abdomen, release the non-invasive artery clamp after clamping for 45 minutes to 1 hour, perform reperfusion for 4 hours, suture the surgical incision with sterile sutures, and then disinfect the surgical area again, put the mouse back into the cage, and give normal feeding and observation; S3. Drug administration test: Administer tripterygium wilfordii derivatives orally or intravenously. 24 hours after administration, evaluate the degree of lung damage by lung tissue sectioning and pathological examination, measure the levels of inflammatory factors (such as IL-6, TNF-α, etc.) in blood and lung tissue, and measure oxidative stress indicators (such as MDA, SOD, etc.) in lung tissue.
2. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In the step a, the concentrated solution of tripterygium wilfordii and silica gel are stirred by a stirrer for 50-80 minutes, and the granular raw material mixed with the solution is dried by a dryer. The eluent is composed of dichloromethane-methanol solution and ammonia water, and the eluent is recovered under reduced pressure by a vacuum decompression recovery machine.
3. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In the step a, the solvent for dissolving the tripterygium wilfordii raw material is ethanol, the heating temperature of the esterification reagent and the solution is 50-150°, which needs to be adjusted according to the reaction conditions, and the esterified derivative is dried by a dryer.
4. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In step b, the culture temperature inside the cell culture incubator is set to 37° C., the carbon dioxide concentration is 5%, and the cell thawing water bath temperature is 37° C.
5. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In step b, the centrifuge tube is centrifuged at a speed of 1000 RPM for 3-5 minutes. The culture medium is a high-glucose DMEM culture medium containing 10% fetal bovine serum. The culture medium needs to be preheated to 37° before use.
6. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In step c, the mice were fasted for 12 hours before modeling, and were allowed to drink water freely before surgery. The surgical area was disinfected with alcohol or iodine, and sterile gauze was placed on the warming pad.
7. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In step c, the length of the abdominal incision of the mouse is about 3-5 cm. The surgical area is covered with sterile gauze soaked in saline, the superior mesenteric artery is clamped with a microvascular clamp, the free intestine is wrapped with sterile gauze, and 37° C. saline is dripped to maintain body temperature and moisture.
8. The protective agent for lung injury caused by intestinal ischemia-reperfusion according to claim 1, characterized in that: In step c, after the operation, mice are given appropriate amounts of antibiotics, and the weight changes, survival rates, and related indicators after intestinal ischemia-reperfusion injury of the mice are recorded.