CN116421664A - Application of coptis detoxification soup in preparation of medicines for treating respiratory tract induced lung injury - Google Patents
Application of coptis detoxification soup in preparation of medicines for treating respiratory tract induced lung injury Download PDFInfo
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Abstract
The invention provides an application of coptis chinensis detoxification soup in preparing a medicament for treating respiratory induced lung injury, belonging to the field of medicines. The coptis detoxification soup disclosed by the invention promotes the polarization of M1/M2 of macrophages and promotes the regression of M2 by regulating and controlling the macrophage non-classical Wnt5a/Fz8 channels, thereby relieving the pulmonary fibrosis after VILI, repairing the lung injury, and has important significance for the prevention and treatment of the VILI, improving the curative effect of positive pressure ventilation and discovering new targets for treatment.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to application of coptis chinensis detoxification soup in preparation of medicines for treating respiratory tract induced lung injury.
Background
Mechanical ventilation is the clinically most important respiratory support means, but from the beginning of the application, it is noted that lung injury is rather further exacerbated in partially treated patients. Ventilator-induced lung injury (VILI) refers to the damage of mechanical ventilation to normal lung tissue or further aggravating the damage of diseased lung tissue, resulting in direct mechanical lung injury, secondary biological injury and oxygen toxicity resulting from increased cross-lung pressure, shear force, and pathology manifests as inflammatory cell infiltration of lung tissue, altered transparent membrane formation and increased vascular permeability. Clinically, the occurrence rate of VILI is as high as 15-50%, which greatly influences the clinical curative effect of mechanical ventilation and even increases the patient's fatality rate. The VILI hazard may persist until after mechanical ventilation is evacuated, and the pulmonary inflammatory response may persist for a longer period of time, inducing abnormal repair of pulmonary tissue, and fibrosis formation. The destruction of lung tissue structure and inflammatory activation by VILI affects lung repair and the onset of fibrosis through a variety of mechanisms, thereby affecting patient recovery of lung function and long-term prognosis. At present, the most widely used clinical application is lung protective ventilation which takes small tidal volume and higher level positive end expiratory pressure as main means, but any positive pressure ventilation strategy cannot avoid the occurrence of VILI, and no symptomatic treatment scheme exists in VILI.
The coptis detoxification soup is a classical representation of a heat-clearing and detoxification method, and is derived from the secret key of the foreign table written by Tang dynasty Wang, and consists of four traditional Chinese medicines of coptis chinensis, phellodendron bark, baical skullcap root and gardenia, and has the effects of clearing heat, purging fire and detoxifying. It has certain curative effect on various diseases, and is not only in accordance with the theory of 'simultaneous treatment of different diseases' in traditional Chinese medicine, but also has the function of regulating systemic immune response and local immune response. At present, no in-vivo and in-vitro study for treating VILI by using coptis chinensis detoxification soup exists.
Disclosure of Invention
Accordingly, the invention aims to provide an application of coptis chinensis detoxification soup in preparing medicines for treating respiratory induced lung injury, so as to relieve lung fibrosis after VILI and repair lung injury.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an application of coptis chinensis detoxification soup in preparing a medicament for treating respiratory induced lung injury.
Preferably, the coptis detoxification soup is one or only one of the active ingredients of the medicine for treating respiratory induced lung injury.
Preferably, the ventilator-induced lung injury includes injury to normal lung tissue or diseased lung tissue caused by mechanical ventilation, and lung injury induced by prolonged inflammatory response of the lung following withdrawal of mechanical ventilation.
Preferably, the damage to normal or diseased lung tissue caused by mechanical ventilation includes direct mechanical lung injury, secondary lung biological injury, and pulmonary type oxygen poisoning.
Preferably, the lung injury induced by the persistence of the inflammatory response of the lung after withdrawal of mechanical ventilation comprises pulmonary fibrosis.
Preferably, the coptis detoxification soup reduces the pulmonary fibrosis area.
Preferably, the coptis detoxification soup promotes the polarization of M1/M2 of macrophages and promotes the regression of M2 type macrophages.
Preferably, the coptis detoxification soup reduces the expression levels of Wnt5a and Fz8 on macrophages.
Preferably, the medicine comprises an effective dose of coptis detoxification soup or a medicine extract in the coptis detoxification soup.
Preferably, pharmaceutically acceptable auxiliary materials are added to prepare powder, decoction, pills, capsules, tablets, granules and oral liquid.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides an application of coptis chinensis detoxification soup in preparing a medicament for treating respiratory induced lung injury. The coptis detoxification soup disclosed by the invention can promote the polarization of M1/M2 of macrophages and promote the regression of M2 macrophages by regulating and controlling the macrophage non-classical Wnt5a/Fz8 channels, thereby relieving the pulmonary fibrosis after VILI and repairing the lung injury, and has important significance for the prevention and treatment of VILI, the improvement of the curative effect of positive pressure ventilation and the discovery of new targets for treatment.
Drawings
Fig. 1: lung tissue fibrosis formation after VILI, a: masson staining examined fibrosis at various time points after VILI withdrawal (blue-green area shows collagen deposition); b: the fibrosis area was counted at each time point after withdrawal, n=6, # p <0.05vs control group, # p <0.05vs 5 days after withdrawal;
fig. 2: lung tissue macrophage infiltration after VILI, a: mac3 immunohistochemical staining detects macrophage infiltration at various time points after VILI withdrawal (tan shows Mac3 positive macrophages); b: counting macrophage infiltration quantity at each time point after withdrawal, wherein n=6, p <0.05vs control group;
fig. 3: effect of coptis detoxification soup on pulmonary fibrosis after VILI, a-C: masson staining examined lung fibrosis between the different groups (blue-green areas show collagen deposition); d: counting the fibrosis areas of each group, wherein n=6;
fig. 4: flow cytometry detection of M1/M2 macrophage subtype distribution in alveolar lavage fluid at various time points after withdrawal of the VILI model, a: flow cytometry F4/80 + CD206 - (M1 type macrophage) and F4/80 + CD206 + (M2 type macrophages); b: proportion statistics of M1, M2 subpopulations (n=6); c: the coptis detoxification soup can advance the reversal time point of the M1/M2 proportion of lung tissue repair (n=6);
fig. 5: effect of coptis detoxification soup on expression level of Wnt5a on macrophages after VILI, a: qRT-PCR (quantitative reverse transcription-polymerase chain reaction) is used for detecting the expression level of mRNA (messenger ribonucleic acid) of a plurality of Wnt family members such as Wnt1, wnt2, wnt3a, wnt5a and the like of a lung tissue of a VILI model; b: qRT-PCR shows that the coptis detoxification soup can reduce the mRNA expression level of Wnt5a in lung tissues; c: immunofluorescence showed that Wnt5a co-localized with macrophage marker F4/80; d: westernblot shows that the coptis chinensis detoxification soup can reduce the expression level of Wnt5a protein in lung tissues;
fig. 6: effect of coptis detoxification soup on expression level of Fz8 on macrophages after VILI, a: qRT-PCR (quantitative reverse transcription-polymerase chain reaction) detection of mRNA (messenger ribonucleic acid) expression level of lung tissue Fz8 of a VILI model; b: qRT-PCR shows that the coptis detoxification soup can reduce the mRNA expression level of lung tissue Fz 8; c: immunofluorescence showed that Fz8 co-localized with macrophage marker F4/80; d: westernblot shows that the coptis detoxification soup can reduce the expression level of Fz8 protein in lung tissues.
Detailed Description
The invention provides an application of coptis chinensis detoxification soup in preparing a medicament for treating respiratory induced lung injury.
The coptis detoxification soup is a classical representation of a heat-clearing and detoxification method, and is derived from the secret key of the foreign table written by Tang dynasty Wang, and consists of four traditional Chinese medicines of coptis chinensis, phellodendron bark, baical skullcap root and gardenia, and has the effects of clearing heat, purging fire and detoxifying.
The coptis detoxification soup disclosed by the invention is prepared from the following raw materials in parts by weight.
In the invention, the respiratory machine induced lung injury comprises injury of normal lung tissue or pathological change lung tissue caused by mechanical ventilation and lung injury induced by lung inflammatory reaction persistence after mechanical ventilation evacuation.
The damage to normal lung tissue or diseased lung tissue caused by mechanical ventilation according to the present invention includes direct mechanical lung injury, secondary lung biological injury and pulmonary oxygen poisoning. Mechanical ventilation causes damage to normal lung tissue, or further exacerbates damage to diseased lung tissue, resulting in increased cross-lung pressure, shear force leading to direct mechanical lung injury, secondary lung biological injury, and pulmonary oxygen toxicity, pathology manifests as alterations characterized by inflammatory cell infiltration of lung tissue, clear membrane formation, and increased vascular permeability.
Pulmonary inflammatory response-mediated lung injury following mechanical ventilation withdrawal described herein includes pulmonary fibrosis. The VILI hazard may persist until after mechanical ventilation is evacuated, and the pulmonary inflammatory response may persist for a longer period of time, inducing abnormal repair of pulmonary tissue, and the formation of pulmonary fibrosis.
The present study found that lung macrophages can be continuously recruited after VILI, activated M1 type macrophages and activated M2 type macrophages in place of activated M1 type macrophages are two subpopulations that are polarized. Early stages of lung tissue injury, i.e., acute inflammatory phase, necrotic apoptotic cells recruit pro-monocytes to aggregate, differentiate into classically activated M1-type macrophages, clear necrotic tissue and express a number of pro-inflammatory cytokines, such as Interleukin-1 beta (IL-1 beta), IL-1, IL-6, interferon gamma (Interferon-gamma), etc. The subsequent inhibition of pro-inflammatory signals, which are more prone to differentiate into activated M2-type macrophages, express high levels of anti-inflammatory cytokines and growth factors such as IL-4, IL-10, transforming growth factor beta (TGF- β), etc., promote epithelial and endothelial cell differentiation and proliferation, restore tissue morphology, and promote cell regeneration and tissue repair; however, if M2 macrophages continue to activate and are accompanied by hypersecretion, inappropriate repair such as alveolar epithelial metaplasia, fibroblast activation, local collagen fiber deposition in the lung, etc. will also occur after VILI, ultimately leading to pulmonary fibrosis, affecting long-term lung function.
In the invention, the coptis detoxification soup is taken as one or only one of the active ingredients, so that the expression level of Wnt5a and Fz8 on macrophages can be effectively reduced, the polarization of M1/M2 of the macrophages is promoted, the regression of M2 type macrophages is promoted, and the pulmonary fibrosis area is reduced. The research of the invention shows that the coptis detoxification soup promotes the polarization of M1/M2 of macrophages by regulating and controlling the macrophage non-classical Wnt5a/Fz8 pathway and promotes the regression of M2 type macrophages, thereby reducing the pulmonary fibrosis after VILI and repairing the lung injury.
In the invention, the prepared medicine for treating respiratory induced lung injury comprises an effective dose of coptis detoxification soup or a medicine active ingredient in the coptis detoxification soup.
In the invention, the coptis chinensis detoxification soup is orally taken. As an implementation mode, the invention is characterized in that coptis chinensis, phellodendron bark, radix scutellariae and gardenia are weighed according to the mass ratio, and the feed liquid ratio is 1: soaking 5-7g/mL in water for 60-120min, preferably 80-100min; soaking, boiling with strong fire, decocting with slow fire for 25-35min, preferably 28-32min, and collecting medicinal liquid; mixing the residues according to a feed liquid ratio of 1: adding water into 3-5g/mL, boiling with strong fire, decocting with slow fire for 20-30min, preferably 24-26min, and collecting medicinal liquid; the two liquid medicines are filtered after being combined, and as an implementation mode, the invention uses a 80-100 mesh filter screen for filtering; decocting the filtered liquid medicine with slow fire, and concentrating until the content of crude drug is 0.5-1g/mL; cooling, packaging, and storing at 4deg.C. The slow fire and the strong fire are Chinese medical nouns, and the normal operation in the field is adopted.
The coptis detoxification soup also comprises the mixture of the raw material extracts. As an implementation mode, the coptis chinensis, the phellodendron bark, the radix scutellariae and the gardenia are weighed according to the mass ratio and then are respectively extracted, and the extracts are taken as the components of the coptis chinensis detoxification soup. The invention does not limit the extraction mode of the extracts of the raw material components in the coptis detoxification soup.
In the medicine for treating the respiratory-induced lung injury, the coptis detoxification soup or the medicine active ingredient in the coptis detoxification soup is taken as the only active ingredient, and the coptis detoxification soup or the medicine active ingredient in the coptis detoxification soup and other active ingredients with the function of treating the respiratory-induced lung injury can be jointly used as the active ingredient.
In the invention, the medicament also comprises pharmaceutically acceptable auxiliary materials, and different dosage forms are prepared by adding the auxiliary materials, wherein the dosage forms comprise powder, decoction, pills, capsules, tablets, granules and oral liquid.
In the present invention, all raw material components are commercially available products well known to those skilled in the art unless specified otherwise.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
This example establishes a large tidal volume induced VILI pulmonary fibrosis model
Male mice of C57/B6J for 6-8 weeks, 18-22g in weight, were selected from Experimental animal technologies, inc., beijing, uighur. The culture was carried out for 7 days before the experiment to adapt to the environment. The room temperature is 22+/-1 ℃, the relative humidity is 50+/-5%, the illumination and darkness are alternated for 12 hours, and the food and water are freely drunk; mice were randomly divided into experimental and control groups.
Experimental group treatment: the mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium, then fixed on an operating table, placed in a supine position, soaked with 75% alcohol, and the anterior cervical skin was cut along the median cervical position, the subcutaneous tissue was separated, the trachea was fully exposed, then 20G venous cannula was inserted into the annular cartilage lower margin and into the trachea, the inner pillow core was pulled out, and the venous cannula and trachea were fixed with silk threads. The small animal breathing machine is connected, and the regulation mode is a capacity control mode. The ventilator parameters were set as follows: tidal volume: 20mL/kg; respiratory rate: 40 times/min; positive end-expiratory pressure (PEEP): 0cmH 2 O;FiO 2 :0.21. mechanical ventilation for 4h, during which anesthesia was maintained and pantoprazole maintained muscle relaxation (2 mg/kg/h). The body temperature of the mice during mechanical ventilation was maintained at 37 ℃ using a heating plate. After mechanical ventilation is finished, the venous cannula is pulled out, the suture is used for suturing, and the neck wound is fixed by using a medical adhesive tape after the iodophor is disinfected.
Control group treatment: the mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital, then fixed on an operating table, placed in supine position, and soaked with 75% alcohol. The anterior cervical skin was incised along the median cervical region, and the subcutaneous tissue was separated tenaciously to fully expose the trachea. Then, a 20G venous cannula is inserted into the trachea at the lower edge of the cricoid cartilage, the inner pillow core is pulled out, and the venous cannula and the trachea are fixed by silk threads. Mice remained spontaneously breathing. For 4h, anesthesia was maintained during which time pantoprazole maintained muscle relaxation (2 mg/kg/h). The body temperature of the mice was maintained at 37 ℃ using a heating plate. After the cannula is completed for 4 hours, the venous cannula is pulled out, the suture is used for suturing, and after the iodophor is sterilized, the neck wound is fixed by using a medical adhesive tape.
Lung tissue sampling was performed on the experimental and control mice at 0d, 1d, 3d, 5d, 7d, 14d and 28d, respectively, after withdrawal. The chest was cut, the lungs of the mice were fully exposed, the portals were found, and the portals were gripped with forceps. Carefully pull the lungs up hard, take the lungs out intact and place in a petri dish. The obtained lung tissue was fixed by soaking overnight in 4% paraformaldehyde at 4deg.C, paraffin sections were prepared by conventional dehydration, hematoxylin-eosin (HE) staining and Masson staining were performed, and lung tissue repair and fibrosis were observed under a microscope, and the results are shown in FIG. 1.
Large area collagen deposition occurs in the lung interstitial mass at day 5 after withdrawal of mice in the experimental group, early fibrosis (Masson stained bluish-green area, indicated by arrow) is shown, progressive exacerbation occurs, and after 14 and 28 days, the fibrosis area is significantly further enlarged.
After withdrawal of the mice 1d, 3d, 5d, 7d, 14d and 28d, the experimental group and the control group mice were subjected to immunohistochemical staining of macrophage marker Mac3 on paraffin sections of lung tissue, and the infiltration of macrophages in the lung tissue was observed, and the results are shown in FIG. 2.
The large amount of macrophage infiltration (immunohistochemical brown yellow region) is visible in the lung interstitium and alveoli at day 3 after the withdrawal of the mice in the experimental group, and the continuous infiltration time is long, and the infiltration of the large amount of macrophage amount can be still detected at day 14 and day 28.
Example 2
The embodiment explores the effect of the coptis detoxification soup on the pulmonary fibrosis of the VILI mouse model
Preparation of coptis detoxification soup: weighing 9g of coptis chinensis, 6g of phellodendron bark, 6g of baical skullcap root and 9g of cape jasmine, adding 180mL of water, soaking for 1 hour, boiling with strong fire, decocting with slow fire for 30 minutes, and pouring out the liquid medicine; adding 120mL of water, boiling with strong fire, and decocting with slow fire for 25 minutes; mixing the two medicinal liquids, filtering with 100 mesh sieve, decocting the filtered medicinal liquid with slow fire, and concentrating to crude drug content of 0.5g/mL; cooling, packaging, and storing at 4deg.C.
Mice were divided into a blank control group, a VILI model control group and a coptis detoxification soup treatment group.
The blank control group is a control group mouse in the example 1, and after the trachea cannula is pulled out, the stomach is irrigated with 0.6mL of water for 14 days continuously;
the experimental group mice in example 1 were randomly divided into two groups; a group of 0.6mL water was infused continuously for 14 days after withdrawal, which was the VILI model control group; a group of coptis chinensis detoxification soup prepared by the embodiment and filled with 0.6mL of stomach for 14 continuous days after the machine is removed is a coptis chinensis detoxification soup treatment group.
Lung tissue sampling was performed on the VILI model control mice at day 5 and day 14 after withdrawal, and lung tissue sampling was performed on the blank control mice and the coptis chinensis detoxification soup treatment mice at day 14 after withdrawal. The lung tissue obtained by the method is fixed, paraffin sections are manufactured through conventional dehydration, HE staining and Masson staining are carried out, lung tissue repair and fibrosis conditions are observed under a microscope, and as a result, the lung fibrosis area of the coptis chinensis detoxification soup group is found to be obviously reduced (as shown in figure 3).
Example 3
The example explores the effect of HUANGLIANJIEDU decoction on macrophage M1/M2 polarization and M2 macrophage resolution of VILI mouse model
The experimental subjects are mice in a blank control group, a VILI model control group and a coptis chinensis detoxification soup treatment group; grouping of mice and intervention into each group of mice was the same as in example 2.
The proportion of M1 and M2 subtype macrophages in the alveolar lavage fluid of mice in the VILI model control group and the coptis detoxification soup treatment group were measured at 0d, 1d, 3d, 5d, 7d and 14d, respectively, after withdrawal.
The detection method comprises the following steps: alveolar lavage fluid macrophages are respectively marked by CD45.2PerCP, F4/80PE and CD206FITC three-color flow fluorescent antibodies, CD45.2PerCP positive cells are taken as a loop gate, F4/80PE positive cells are macrophages, and F4/80 + CD206 - The cells were M1 type macrophages (upper left quadrant), F4/80 + CD206 + The cells were M2 type macrophages (upper right quadrant), and the flow cytometer automatically detected the cell proportions in each quadrant (results are shown in FIG. 4).
Flow cell detection results show that after VILI is generated by VILI model control mice, the quantity of M1 type macrophages is dominant within 0-5 days after the mice are removed, the proportion of M2 type is gradually increased, and the proportion of M1/M2 type is reversed on the 7 th day and is continued until the 14 th day (fig. 4A); counting the proportion of M1 and M2 type macrophages at each time point after the withdrawal (figure 4B), and finding that the time point when the proportion of M1/M2 in lung tissue repair is reversed is delayed to 7 days after the withdrawal;
the detection result of mice in the golden thread detoxification soup treatment group shows that the golden thread detoxification soup can inhibit the massive infiltration of macrophages, and can advance the time point when the M1/M2 proportion of lung tissue repair is reversed to the 5 th day after the respiratory machine is evacuated (fig. 4C).
The results show that the coptis detoxification soup can promote the polarization of the VILI model macrophage M1/M2 and promote the elimination of M2.
Example 4
The present example explores the effect of Huanglian Jiedu Tang on the expression level of Wnt5a on VILI mouse model macrophages
The experimental subjects are mice in a blank control group, a VILI model control group and a coptis chinensis detoxification soup treatment group; grouping of mice and intervention into each group of mice was the same as in example 2.
On day 14 after withdrawal, lung tissues of mice from the blank and VILI model control groups were collected, frozen and ground with liquid nitrogen, RNA was extracted with trizol, reverse transcribed, and qPCR was performed to detect expression levels of Wnt family member mRNA. The results show that the expression levels of a plurality of members such as Wnt1, wnt2, wnt3a, wnt5a and the like in the lung tissues of the mice in the VILI model control group are up-regulated, and the expression of Wnt10a is down-regulated. Wherein Wnt5A up-regulation was most pronounced, fold change was more than 5-fold (fig. 5A).
On day 14 after withdrawal, mice from the blank control group, the VILI model control group and the coptis detoxification soup treatment group were tested respectively:
1) Directly taking lung tissues, performing liquid nitrogen freeze grinding, extracting RNA by using trizol, and detecting the mRNA expression level of Wnt5a by using qPCR after reverse transcription. The results showed that mRNA expression levels of Wnt5a were significantly reduced in the lung tissue of the coptis detoxification soup treatment group (fig. 5B).
2) After the mice are anesthetized, the left atrium is cannulated, circulating blood is removed by PBS, lung tissue is obtained after the 4% paraformaldehyde is fully perfused, and the mice are soaked in the 4% paraformaldehyde at the temperature of 4 ℃ for overnight. Then placing in 30% sucrose solution, dewatering at 4deg.C for more than 24 hr until the bottom is completely settled, embedding O.C.T, and freezing and slicing. Slice tissues were subjected to tissue immunofluorescence experiments, macrophages were labeled with anti-F4/80 antibody, and Wnt5a was labeled with anti-Wnt5a antibody. The results showed that Wnt5a co-localized with F4/80, indicating that Wnt5a was predominantly distributed on macrophages (fig. 5C).
3) Directly taking lung tissues, performing liquid nitrogen freeze grinding, dissolving by using RIPA lysate, performing ultrasonic crushing, extracting protein, and detecting the expression level of Wnt5a protein by using western blot. The results showed that protein expression levels of Wnt5a were significantly reduced in the coptis detoxification soup treatment group (fig. 5D).
The results show that the coptis detoxification soup can obviously reduce the expression level of Wnt5a on VILI model macrophages.
Example 5
The present example explores the effect of Huanglian Jiedu Tang on the expression level of Fz8 on VILI mouse model macrophages
The experimental subjects are mice in a blank control group, a VILI model control group and a coptis chinensis detoxification soup treatment group; grouping of mice and intervention into each group of mice was the same as in example 2.
On days 3, 5, 7, and 14 after withdrawal, mouse lung tissues of the blank control group and the VILI model control group were collected, RNA was extracted with trizol after liquid nitrogen freeze-milling, and qPCR was performed after reverse transcription to detect the expression level of mRNA of Frizzled (Fz) receptor family member Fz8. The results show that mRNA expression levels of VILI model lung tissue Fz8 were significantly up-regulated (fig. 6A).
On day 14 after withdrawal, mice from the blank control group, the VILI model control group and the coptis detoxification soup treatment group were tested respectively:
1) Lung tissue was directly harvested and liquid nitrogen cryo-milled, RNA was extracted using trizol, reverse transcribed and mRNA expression levels of Fz8 were detected using qPCR. The results showed that mRNA expression level of lung tissue Fz8 was significantly reduced in coptis detoxification soup treatment group (fig. 6B).
2) After the mice are anesthetized, the left atrium is cannulated, circulating blood is removed by PBS, lung tissue is obtained after the 4% paraformaldehyde is fully perfused, and the mice are soaked in the 4% paraformaldehyde at the temperature of 4 ℃ for overnight. Then placing in 30% sucrose solution, dewatering at 4deg.C for more than 24 hr until the bottom is completely settled, embedding O.C.T, and freezing and slicing. Tissue immunofluorescence experiments were performed on the sectioned tissues, macrophages were labeled with anti-F4/80 antibody, and Fz8 was labeled with anti-Fz8 antibody. The results showed that Fz8 co-localizes with F4/80, indicating that Fz8 is predominantly distributed on macrophages (FIG. 6C).
3) Directly taking lung tissues, performing liquid nitrogen freeze grinding, dissolving by using RIPA lysate, performing ultrasonic crushing, extracting protein, and detecting the expression level of Fz8 protein by using western blot. The results showed that protein expression level of coptis detoxification soup treatment group Fz8 was significantly reduced (fig. 6D).
The results show that the coptis detoxification soup can obviously reduce the expression level of Fz8 on VILI model macrophages.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. Application of Coptidis rhizoma decoction for removing toxic substances in preparing medicine for treating respiratory tract injury is provided.
2. The use according to claim 1, wherein the coptis detoxification soup is one or only one of the active ingredients of the medicament for treating ventilator-induced lung injury.
3. The use of claim 1, wherein the ventilator-induced lung injury comprises injury to normal or diseased lung tissue caused by mechanical ventilation, and lung injury induced by a prolonged inflammatory response of the lung following withdrawal of mechanical ventilation.
4. The use according to claim 3, wherein the damage to normal or diseased lung tissue caused by mechanical ventilation comprises direct mechanical lung injury, secondary lung biological injury and pulmonary type oxygen poisoning.
5. The use according to claim 3, wherein the lung injury induced by the persistence of the inflammatory response of the lung following evacuation of mechanical ventilation comprises pulmonary fibrosis.
6. The use according to any one of claims 3-5, wherein the coptis detoxification soup reduces the area of pulmonary fibrosis.
7. The use according to any one of claims 3-5, wherein the coptis detoxification soup promotes M1/M2 polarization of macrophages, promoting M2 type macrophage regression.
8. The use according to any one of claims 3 to 5, wherein the coptis detoxification soup reduces the expression levels of Wnt5a and Fz8 on macrophages.
9. The use according to claim 1, wherein the medicament comprises an effective dose of coptis detoxification soup or a medicament extract in coptis detoxification soup.
10. The use according to claim 9, wherein the powder, decoction, pill, capsule, tablet, granule and oral liquid are prepared by adding pharmaceutically acceptable auxiliary materials.
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刘保光等: "黄连解毒汤作用机制研究进展", 中医学报, vol. 37, no. 09, 30 September 2022 (2022-09-30), pages 1861 - 1868 * |
李翀等: "黄连解毒汤对脓毒症模型小鼠肺损伤的保护作用 及其机制研究", 江苏中医药, vol. 50, no. 03, 31 March 2018 (2018-03-31), pages 79 - 82 * |
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