CN114949034A - Application of perilla frutescens or extract thereof in preparation of medicine for preventing and treating acute pneumonia - Google Patents

Application of perilla frutescens or extract thereof in preparation of medicine for preventing and treating acute pneumonia Download PDF

Info

Publication number
CN114949034A
CN114949034A CN202210511606.6A CN202210511606A CN114949034A CN 114949034 A CN114949034 A CN 114949034A CN 202210511606 A CN202210511606 A CN 202210511606A CN 114949034 A CN114949034 A CN 114949034A
Authority
CN
China
Prior art keywords
extract
perilla frutescens
medicine
lung
perilla
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202210511606.6A
Other languages
Chinese (zh)
Other versions
CN114949034B (en
Inventor
田雷
孙瑾
姜晓茜
李晔
郭善成
赵志业
陈昱旭
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kunming University of Science and Technology
Original Assignee
Kunming University of Science and Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kunming University of Science and Technology filed Critical Kunming University of Science and Technology
Priority to CN202210511606.6A priority Critical patent/CN114949034B/en
Publication of CN114949034A publication Critical patent/CN114949034A/en
Application granted granted Critical
Publication of CN114949034B publication Critical patent/CN114949034B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/53Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00Drugs for disorders of the respiratory system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/35Extraction with lipophilic solvents, e.g. Hexane or petrol ether
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/53Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Botany (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Pulmonology (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Pain & Pain Management (AREA)
  • Rheumatology (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses an application of perilla frutescens or an extract thereof in preparing a medicine for preventing and treating acute pneumonia, wherein the medicine is a medicine for inhibiting lung tissue bronchoalveolar inflammatory cell infiltration and protein exudation, inhibiting expression of related inflammatory factors in lung parts, inhibiting expression of related inflammatory factors in plasma and improving pathological damage caused by pulmonary inflammatory reaction. The perilla frutescens extract can effectively inhibit lung tissue bronchoalveolar inflammatory cell infiltration and protein exudation of an acute lung injury model mouse under a lower dose, inhibit expression of related inflammatory factors in lung and plasma of an inflammation model mouse, and effectively improve pathological damage caused by inflammatory reaction of the lung of the mouse; the raw materials of the invention are from natural products, and the invention has convenient collection and high safety.

Description

Application of perilla frutescens or extract thereof in preparation of medicine for preventing and treating acute pneumonia
Technical Field
The invention relates to the technical field of medicines, in particular to application of perilla frutescens or an extract thereof in preparing a medicine for preventing and treating acute pneumonia.
Background
An ideal model for researching the inflammatory reaction of the lung is an animal acute lung injury model, and although researchers deeply research the acute lung injury in laboratories and clinics, the lethality rate is still very high. In addition, the current western medicines for treating acute lung injury have large side effects and are easy to generate drug resistance. In order to prevent or treat the acute pneumonia, a safe and effective medicine for preventing or treating the acute pneumonia is urgently sought from natural products.
Eastern perilla (elsholtziabodinieri vaniot) is a perennial herb of the genus elsholtzia of the family labiatae, and is mainly distributed in parts of Yunnan, Guizhou and Gansu. The perilla frutescens is a traditional plant used for both food and medicine, namely camellia sinensis, pine tea and the like in Yunnan, and branches and leaves of the perilla frutescens are soaked in water to serve as herbal tea for drinking in a plurality of minority regions, so that the perilla frutescens has the effects of clearing heat and reducing internal heat, and is a natural health-care beverage. In addition, the perilla frutescens has pungent property and is used as a medicine by whole herbs, which are recorded in national Chinese herbal medicine assembly and Yunnan Chinese herbal medicine, and has the efficacies of relieving fever, easing pain, stopping vomiting, diminishing inflammation, resisting virus and the like. It is reported that the Elsholtzia bodinieri Vahl is rich in triterpenes, aromatic oils, flavonoids, etc., and the extract thereof has antioxidant, antifatigue and antiviral activities, and the Elsholtzia bodinieri Vahl extract is safe without mutagenic effects. However, at present, no patent literature reports about the anti-inflammatory research of the perilla frutescens extract at home and abroad. The research related to the patent is funded by national science fund (approval number: 32160106).
Disclosure of Invention
In order to solve the technical problems, the invention provides a new application of the perilla frutescens or the extract thereof in the aspect of medicine, and the perilla frutescens or the extract thereof is applied to the preparation of a new medicine for preventing and/or treating acute pneumonia.
In order to achieve the purpose, the invention adopts the following technical scheme:
the first purpose of the invention is to provide an application of the perilla frutescens or the extract thereof in preparing a medicine for preventing and treating acute pneumonia, wherein the medicine is used for inhibiting infiltration and protein exudation of bronchoalveolar inflammatory cells of lung tissues, inhibiting expression of related inflammatory factors in lung parts, inhibiting expression of related inflammatory factors in plasma and improving pathological damage caused by pulmonary inflammatory reaction.
Further, the perilla frutescens extract is prepared by the following method:
(1) pulverizing Elsholtzia bodinieri Van to powder, and decolorizing Elsholtzia bodinieri Van powder: adding petroleum ether, performing suction filtration after ultrasonic treatment, and performing rotary evaporation to recover the petroleum ether to obtain the perilla frutescens residue;
(2) airing the residues of the east perilla in the step (1), adding 80% methanol solution, carrying out ultrasonic extraction, and carrying out suction filtration to obtain east perilla filter residues;
(3) adding 80% methanol solution into the east perilla filter residue in the step (2) to perform ultrasonic extraction for 2-3 times, and combining the extracting solutions; rotary distilling the extract to recover methanol until a water phase is obtained; and continuously performing rotary evaporation and concentration on the water phase to obtain a concentrated solution, pre-freezing the concentrated solution in a refrigerator at the temperature of-20 ℃, and freeze-drying by using a freeze dryer to obtain the perilla frutescens 80% methanol solution extract (freeze-dried powder).
Further, in the step (1), the mass-to-volume ratio of the perilla frutescens powder to the petroleum ether is 1g:10 mL.
Further, the ultrasound in the step (1) is ultrasound for 30-35 min under the power of 60-70 w; the rotary evaporation for recovering petroleum ether is carried out for 3 times.
Further, in the step (2) and the step (3), the mass-to-volume ratio of the east perilla residue to the methanol solution and the mass-to-volume ratio of the east perilla residue to the methanol solution are both 1g:10 mL.
Further, the ultrasonic extraction in the step (2) and the step (3) is performed for 30-35 min under the power of 300-320 w.
In the invention, the concentration of the perilla frutescens extract in the medicine for preventing and treating acute pneumonia is preferably 200 mg/kg-600 mg/kg. The invention has no special requirements on the dosage form of the medicine, the auxiliary materials of the medicine and the preparation method of the medicine, and the invention can adopt the methods well known by the technical personnel in the field. The new medicine for preventing and/or treating acute pneumonia is prepared by using the extract of the east perilla as an active ingredient, is used for preparing the medicine for preventing and treating acute pneumonia, and can be added with one or more auxiliary materials acceptable in pharmaceutical preparations or compounded with other active ingredients to play a role in synergistically resisting acute pneumonia.
The invention carries out series research on the perilla frutescens or the extractive thereof for preventing and treating acute pneumonia. The perilla frutescens extract is proved to be capable of remarkably inhibiting infiltration of lung tissue bronchoalveolar inflammatory cells and protein exudation; can obviously inhibit the expression of relevant inflammatory factors in the lung and plasma of an inflammation model mouse; can effectively improve pathological damage caused by inflammatory reaction of the lung of the mouse; the lung inflammation of lipopolysaccharide-induced acute lung injury model mice is effectively improved and the lung function of the lipopolysaccharide-induced acute lung injury model mice is improved, and the effect on preventing or treating acute pneumonia is almost equivalent to that of a blank control group. The perilla frutescens or the extract thereof provided by the invention has a remarkable effect on preventing and treating acute pneumonia, and the perilla frutescens plant is wide in distribution, high in safety, low in price and easy to obtain raw materials; the perilla frutescens or the extract thereof has good development and application prospects in preparation of the medicine for preventing and treating acute pneumonia.
In summary, compared with the prior art, the invention has the advantages that:
1. the invention explores a new medical application of the perilla frutescens extract.
2. Experiments show that the perilla frutescens extract can effectively inhibit lung tissue bronchoalveolar inflammatory cell infiltration and protein exudation of an acute lung injury model mouse under a lower dose, inhibit expression of related inflammatory factors in lungs and plasma of an inflammation model mouse, and effectively improve pathological damage caused by lung inflammatory reaction of the mouse; the raw materials of the invention are from natural products, and the invention has convenient collection and high safety.
3. The perilla frutescens extract has simple extraction process and low cost, and is suitable for industrial production and market popularization and application.
Drawings
FIG. 1 is a diagram showing the results of liquid phase detection of Elsholtzia bodinieri prepared in example 1 of the present invention;
FIG. 2 is a bar graph of the effect of Elsholtzia bodinieri extract on mouse bronchoalveolar lavage fluid cells and proteins, wherein panel A shows the number of bronchoalveolar lavage fluid cells; panel B shows bronchoalveolar lavage fluid protein levels; c is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone);
FIG. 3 is a graph showing the expression level of the lung inflammatory factor in mice of the present invention, in which FIG. A shows the expression level of IL-6 in lung tissue of mice; FIG. B shows the expression level of IL-1. beta. in lung tissue; FIG. C shows the expression level of MPO in lung tissue; c is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone);
FIG. 4 is a graph showing the expression level of inflammatory factors in the plasma of mice in accordance with the present invention, wherein FIG. A shows the expression level of IL-6 in the plasma of mice; FIG. B shows the expression level of IL-1. beta. in mouse plasma; FIG. C shows the expression level of NO in mouse plasma; c is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone);
FIG. 5 is a graph of HE staining of a lung tissue section of a mouse of the present invention, wherein Control is a blank Control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM was positive group (dexamethasone).
Detailed Description
The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without any inventive step, are within the scope of the present invention. Techniques, methods, and apparatus known to one of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate. In addition, the technical features involved in the different embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Example 1
Preparation of the perilla frutescens extract of this example:
the Elsholtzia bodinieri Hayata is collected from Yunnan Wenshan, and is naturally dried and pulverized to obtain the Elsholtzia bodinieri Hayata dry powder. Weighing 50g of dry powder of the whole herb of the east perilla, adding petroleum ether, performing ultrasonic treatment for 30min under the power of 60w, wherein the mass-volume ratio of the dry powder of the east perilla to the petroleum ether is 1g:10mL, performing suction filtration, performing rotary evaporation and recovery on part of the petroleum ether, repeating the process for 3 times, and keeping the residue of the east perilla. Air drying the decolorized residues of the east perilla, adding 80% methanol, performing ultrasonic treatment at 300w power for 30min, wherein the mass volume ratio of the east perilla to the methanol is 1g:10mL, performing suction filtration, adding 80% methanol into the obtained residues of the east perilla, performing ultrasonic treatment at 300w power for 30min, and repeating the steps for 3 times; then extracting by rotary evaporation to obtain only water phase in a rotary evaporation bottle, recovering methanol, continuously carrying out rotary evaporation and concentration on the water phase, then prefreezing in a refrigerator at-20 ℃, and finally freeze-drying by a freeze dryer to obtain the perilla frutescens 80% methanol extract (freeze-dried powder).
The main chemical components of the Elsholtzia bodinieri Van 80% methanol extract were qualitatively analyzed by UPLC-MS/MS technique, and the analysis results are shown in FIG. 1.
As can be seen from fig. 1: the 9 main chemical components of the eastern perilla 80% methanol extract are quinic acid (1), chlorogenic acid (2), apigenin (3), Luteolin-7-O-rutinoside (4), mangiferin (5), Luteolin (6), cosmosin (7), rosmarinic acid (8), and eridol 7-O- (6' -3, 4-dihydroxycinnamyl) -beta-D-glucopyranoside (9).
The invention provides application of the perilla frutescens extract obtained by the preparation method in preparation of a medicine for treating acute pneumonia.
The invention also provides application of the perilla frutescens extract in preparing a medicine for preventing and treating acute pneumonia by inhibiting lung tissue bronchoalveolar inflammatory cell infiltration and protein exudation.
The invention also provides application of the perilla frutescens extract in preparing a medicine for preventing and treating acute pneumonia by inhibiting expression of related inflammatory factors in lung.
The invention also provides application of the perilla frutescens extract in preparing a medicine for preventing and treating acute pneumonia by inhibiting expression of relevant inflammatory factors in blood plasma.
The invention also provides application of the perilla frutescens extract in preparing a medicine for preventing and treating acute pneumonia by improving pathological injury caused by pulmonary inflammatory reaction.
Example 2
This example is the evaluation of the activity of the extract of perilla frutescens for preventing and treating acute pneumonia:
2.1 establishment and Experimental Process of acute Lung inflammation mouse model
C57BL/6J male mice (8 weeks old, weighing about 20 g) were used as subjects, and 15 mice were administered to each group. Acute lung inflammatory reaction is induced by nasal LPS (0.5mg/kg), a mouse acute lung injury model is constructed, and modeling is carried out for 12 hours. The prevention and treatment effect of the sage on LPS-induced acute lung injury mice is evaluated by the sage eastern 80% methanol extract obtained in the gastric lavage example 1; mice were acclimated for 7 days in standard environment after purchase and mice were fed standard chow. The specific experimental procedures and groups are as follows:
blank control group: administering purified water with the same volume as the stomach;
model group: injecting LPS (0.5mg/kg) into a pipette nose according to the weight, and molding for 12 hours;
low dose group of sage dong: continuously administering 200mg/kg of Elsholtzia bodinieri (Vat.) Kuntze 80% methanol extract for 7 days, and molding with LPS for 24 hr before dissection;
the high dose group of sage dong: continuously administering 600mg/kg of 80% methanol extract of Elsholtzia bodinieri (L.) Druce after 7 days of gastric lavage, and molding with LPS for 24h before dissection;
positive control group: performing intragastric administration for 1h before molding, administering 10mg/kg dexamethasone, and then performing nasal infusion of LPS (LPS) (0.5mg/kg) by a pipette according to body weight for 24h after molding;
2.2 testing the effect of active ingredients on the number of bronchoalveolar lavage fluid cells and protein content of model mice:
2.2.1 taking each group of mice, performing tracheal intubation, irrigating bronchoalveolar with precooled PBS, taking lavage fluid, centrifuging for 5 minutes at 4 ℃ at 1500 revolutions, collecting supernatant, and detecting the content of total protein by using a BCA kit; the remaining cells were resuspended in pre-cooled PBS, stained with a complex Richmson stain (Wright-Giemsa) for 4 minutes, washed by centrifugation to remove the stain, and immediately the macrophages and neutrophils were counted separately under the microscope using a hemocytometer.
2.2.2 results of the experiment:
effect of eastern perilla extract on bronchoalveolar lavage fluid cell number and total protein content of model mice:
the alveolar lavage fluid test can be used to evaluate the inflammatory response stage of interstitial lung disease and diagnose lung disease. By analyzing the cell number and the total protein content of the bronchoalveolar lavage fluid, the method can be used for analyzing and detecting the disease condition of certain respiratory diseases, and particularly plays a very important role in researching pathogenesis. When lung infection and inflammation occur, alveolar inflammatory cell infiltration occurs, the number of alveolar lavage fluid cells is obviously increased, bronchoalveolar cell proteins of lung tissues are exuded, and the total protein content of alveolar lavage fluid is also increased.
The invention evaluates the action condition of the perilla frutescens extract on acute lung injury model mice by the number of bronchoalveolar lavage fluid cells and the total protein content of each treated mouse, and the result is shown in figure 2, wherein a graph A shows the number of bronchoalveolar lavage fluid cells, and a graph B shows the amount of bronchoalveolar lavage fluid protein. C is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone); denotes p < 0.05; denotes p < 0.01.
2.3 detecting the influence of the active ingredients on the expression of the lung tissue inflammatory factor and related proteins of the model mouse:
2.3.1 taking each group of mice, taking lung tissues by aseptic technique, washing with aseptic precooled PBS, taking 100mg of lung tissues, adding 900 mu l of aseptic precooled PBS, homogenizing at 4 ℃, centrifuging, taking supernate, and detecting the expression conditions of IL-6, IL-1 beta and MPO in the lung tissues of each group of mice by using a corresponding kit.
2.3.2 results of the experiment
The inflammatory factors refer to various cytokines involved in inflammatory reaction, and when the lung is stimulated by the outside, the lung cells can generate immune response, secrete a large amount of inflammatory factors and even initiate stronger inflammatory reaction. Typical inflammatory factors include interleukin 6(IL-6), interleukin 1 beta (IL-1 beta), Myeloperoxidase (MPO), and the like. IL-6 is a multi-potent cytokine for proinflammatory and immune regulation, has an inflammation amplification effect, and is involved in the occurrence and development of diseases and the inflammatory response and immune response of organisms. The damage of IL-6 to lung is mainly caused by inflammation caused by lung infection and oxidative stress stimulating local macrophages of lung tissue to activate and promote IL-6 secretion, while IL-6 can activate the aggregation of neutrophils at the inflammation part, thereby releasing a large amount of enzyme and oxygen free radicals, and causing the increase of lung capillary permeability and the degeneration and necrosis of lung epithelial cells. IL-1. beta. is a signal to initiate inflammation. MPO is a heme protein, abundant in neutrophils, synthesized in the bone marrow before entry into the circulation by granulocytes and stored in azure granules. MPO can kill microorganisms in phagocytes by catalyzing and oxidizing chloride ions to generate hypochlorous acid, destroy various target substances, and play a role in various aspects such as organism generation, inflammatory reaction regulation and the like.
According to the invention, the influence of the Elsholtzia bodinieri extract on the acute lung injury mice is examined by detecting the expression conditions of IL-6, IL-1 beta and MPO in the lung tissues of each group of mice by using a corresponding kit, and the effect is shown in figure 3. Wherein Panel A shows the expression level of IL-6 in lung tissue; FIG. B shows the expression level of IL-1. beta. in lung tissue; FIG. C shows the expression level of MPO in lung tissue. C is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone); denotes p < 0.05.
The results in fig. 3 show that the perilla frutescens extract can significantly reduce the expression of IL-6 (p <0.05), the expression of IL-1 β (p <0.05) and the expression of MPO (p <0.05) in lung tissues of mice as a model of inflammation. Therefore, the perilla frutescens extract can obviously inhibit the expression of the relevant inflammatory factors of the lung of the mouse model with inflammation.
2.4 testing the effect of the active principle on the inflammatory factors in the plasma of model mice:
2.4.1 taking each group of mice, collecting blood by anticoagulation tube eyeballs, centrifuging for 5 minutes at 2500 rpm, taking supernatant plasma, and detecting the expression conditions of IL-6, IL-1 beta and NO in the plasma of each group of mice by using a corresponding kit.
2.4.2 results of the experiment
NO has proinflammatory effect, and at an inflammation part, NO acts on vascular smooth muscle cells to increase cGMP level of the vascular smooth muscle cells, so that blood vessels are dilated, permeability is increased, inflammatory mediators and pain substances can reach the action part, and infiltration of monocytes to the inflammation part is increased in inflammatory reaction. In the course of the pathogenesis of immune inflammation, elevated levels of NO in vivo can affect the pathological process from multiple pathways.
The invention inspects the influence of the perilla alcohol extract on the mice with acute lung injury by detecting the expression conditions of IL-6, IL-1 beta and NO in the blood plasma of each group of mice by using a corresponding kit, and the effect is shown in figure 4. Wherein Panel A shows the expression level of IL-6 in plasma; panel B shows the amount of IL-1. beta. expression in plasma; panel C shows the amount of NO expressed in plasma. C is blank control group; model is LPS induction Model group; EBEL is low dose gavage group (200 mg/kg); EBEH was high dose gavage group (600 mg/kg); DXM as positive group (dexamethasone); denotes p <0.05, denotes p < 0.01.
The results in fig. 4 show that the extract of perilla frutescens can significantly reduce the expression of IL-6 (, p <0.01), the expression of IL-1 β (, p <0.05) and the expression of NO (, p <0.01) in the plasma of mice model for inflammation. Therefore, the perilla frutescens extract can obviously inhibit the expression of related inflammatory factors in the plasma of an inflammation model mouse.
2.5 testing the effect of active ingredients on the lung tissue structure of model mice:
2.5.1 taking each group of mice, taking lung tissues by aseptic technique, washing with sterile precooled PBS, fixing to prepare paraffin sections, and observing the change condition of the tissue structure under a microscope after HE staining.
The preparation method of the paraffin section comprises the following steps:
(1) material taking: fresh lung tissue was fixed with 10% formalin for over 24 h. Taking out the lung tissue from the stationary liquid, flattening the tissue of a target part in a fume hood by using a scalpel, and placing the trimmed tissue and a corresponding label in a dehydration box;
(2) dehydrating and wax dipping: and (5) putting the dehydration box into a dehydration machine for dehydration by gradient alcohol in sequence. The method comprises the following steps of 1, dissolving paraffin I at 65 ℃ for 1h, dissolving paraffin II at 65 ℃ for 1h, dissolving paraffin III at 85% for 2h, dissolving 90% alcohol for 2h, dissolving 95% alcohol for 1h, dissolving absolute ethanol I for 30min, dissolving absolute ethanol II for 30min, dissolving alkylbenzene for 5-10min, xylene I for 5-10min, xylene II for 5-10 min;
(3) embedding: embedding the wax-soaked tissue in an embedding machine. Firstly, molten wax is put into an embedding frame, tissues are taken out from a dehydration box and put into the embedding frame according to the requirements of an embedding surface before the wax is solidified, and corresponding labels are attached. Freezing at-20 deg.C, cooling, solidifying wax, taking out the wax block from the embedding frame, and trimming the wax block;
(4) slicing: placing the trimmed wax block in a paraffin slicer for slicing, wherein the thickness of the wax block is 4 mu m; the slices float on a spreading machine at 40 ℃ warm water to flatten the tissues, the glass slides pick up the tissues, the slices are baked in a 60 ℃ oven, and the slices are taken out and stored at normal temperature for later use after being baked by water, dried and waxed.
HE staining method was as follows:
(1) paraffin section dewaxing to water: sequentially placing the slices into xylene I20 min-xylene II 20 min-absolute ethyl alcohol I5 min-absolute ethyl alcohol II 5 min-75% alcohol 5min, and washing with tap water;
(2) hematoxylin staining: staining the slices in hematoxylin staining solution for 3-5min, washing with tap water, differentiating the differentiation solution, washing with tap water, returning blue to the blue solution, and washing with running water;
(3) eosin staining: the slices are dehydrated for 5min respectively by 85 percent and 95 percent gradient alcohol, and are dyed for 5min in eosin dye solution.
(4) Dewatering and sealing: placing the slices in anhydrous ethanol I5 min-anhydrous ethanol II 5 min-anhydrous ethanol III 5 min-xylene I5 min-xylene II 5min for transparency, and sealing with neutral gum;
(5) microscopic examination and image acquisition and analysis.
2.5.2 results of the experiment
The HE staining was used in this experiment to observe the lung tissue structure of mice, and the results are shown in FIG. 5.
As can be seen from the results in fig. 5, the perilla frutescens extract can significantly reduce inflammatory exudates between bronchoalveolar cells, significantly reduce infiltration of tissue inflammatory cells, and maintain integrity of bronchoalveolar epithelial cells. The results show that the perilla frutescens extract can obviously improve pathological injury caused by lung inflammatory reaction of mice.
The preparation method of the perilla frutescens extract comprises the following steps: adding petroleum ether under 70w power for 35min, or adding petroleum ether under 65w power for 33min, and ultrasonic extracting with 80% methanol under 320w power for 35min or 32min, and mixing the rest with the above extract to obtain Coleus blumei extract for preventing and treating acute pneumonia.
In conclusion, the perilla frutescens extract can obviously inhibit the infiltration of lung tissue bronchoalveolar inflammatory cells and protein exudation; can obviously inhibit the expression of relevant inflammatory factors in the lung and plasma of an inflammation model mouse; can effectively improve pathological injury caused by inflammatory reaction of the lung of the mouse. The perilla frutescens extract provided by the invention has a remarkable effect on preventing and treating acute pneumonia, is prepared from natural products, has high safety and low cost, and is suitable for industrial production and market popularization and application.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
Although embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are exemplary and not to be construed as limiting the present invention, and that those skilled in the art may make variations, modifications, substitutions and alterations within the scope of the present invention without departing from the spirit and scope of the present invention.

Claims (6)

1. The application of the perilla frutescens or the extract thereof in preparing the medicine for preventing and treating acute pneumonia is characterized in that the medicine is a medicine for inhibiting infiltration and protein exudation of lung tissue bronchoalveolar inflammatory cells, inhibiting expression of related inflammatory factors in lung, inhibiting expression of related inflammatory factors in plasma and improving pathological damage caused by lung inflammatory reaction.
2. The use of the perilla frutescens or the extract thereof in the preparation of a medicament for preventing and treating acute pneumonia according to claim 1, wherein the perilla frutescens extract is prepared by the following method:
(1) pulverizing Elsholtzia bodinieri Van to powder, decolorizing the Elsholtzia bodinieri Van powder, adding petroleum ether, performing ultrasonic filtration, and performing rotary evaporation to recover the petroleum ether to obtain Elsholtzia bodinieri Van residue;
(2) airing the residues of the east perilla in the step (1), adding 80% methanol solution, carrying out ultrasonic extraction, and carrying out suction filtration to obtain east perilla filter residues;
(3) adding 80% methanol solution into the east perilla filter residue in the step (2), performing ultrasonic extraction for 2-3 times, and combining the extracting solutions; rotary distilling the extract to recover methanol until a water phase is obtained; and continuously performing rotary evaporation and concentration on the water phase to obtain a concentrated solution, pre-freezing the concentrated solution in a refrigerator at the temperature of-20 ℃, and performing freeze-drying by using a freeze dryer to obtain the perilla frutescens 80% methanol solution extract.
3. The application of the perilla frutescens or the extract thereof in preparing the medicine for preventing and treating acute pneumonia in claim 2 is characterized in that in the step (1), the mass-to-volume ratio of the perilla frutescens powder to the petroleum ether is 1g:10 mL.
4. The use of the perilla frutescens or the extract thereof in the preparation of the medicine for preventing and treating acute pneumonia according to claim 2, wherein the ultrasound in the step (1) is performed for 30-35 min at a power of 60-70 w; the rotary evaporation for recovering petroleum ether is carried out for 3 times.
5. The application of the perilla frutescens or the extract thereof in preparing the medicine for preventing and treating acute pneumonia in claim 2 is characterized in that in the step (2) and the step (3), the mass-to-volume ratio of the perilla frutescens residue to the methanol solution and the mass-to-volume ratio of the perilla frutescens residue to the methanol solution are both 1 g/10 mL.
6. The use of the perilla frutescens or the extract thereof in the preparation of the drug for preventing and treating acute pneumonia according to claim 2, wherein the ultrasonic extraction in the step (2) and the ultrasonic extraction in the step (3) are both ultrasonic extraction at power of 300-320w for 30-35 min.
CN202210511606.6A 2022-05-12 2022-05-12 Application of perilla extract in preparing medicine for preventing and treating acute pneumonia Active CN114949034B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202210511606.6A CN114949034B (en) 2022-05-12 2022-05-12 Application of perilla extract in preparing medicine for preventing and treating acute pneumonia

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202210511606.6A CN114949034B (en) 2022-05-12 2022-05-12 Application of perilla extract in preparing medicine for preventing and treating acute pneumonia

Publications (2)

Publication Number Publication Date
CN114949034A true CN114949034A (en) 2022-08-30
CN114949034B CN114949034B (en) 2023-11-07

Family

ID=82981289

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202210511606.6A Active CN114949034B (en) 2022-05-12 2022-05-12 Application of perilla extract in preparing medicine for preventing and treating acute pneumonia

Country Status (1)

Country Link
CN (1) CN114949034B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116392518A (en) * 2023-04-19 2023-07-07 昆明理工大学 Application of perilla and extract thereof in preparing medicine for treating chronic pressure brain lesions

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101156889A (en) * 2004-04-10 2008-04-09 云南省药物研究所 Application of east purple common perilla extractive
CN113398222A (en) * 2020-03-17 2021-09-17 云南中医药大学 National medicine composition for exorcising evil spirits and preventing epidemic

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101156889A (en) * 2004-04-10 2008-04-09 云南省药物研究所 Application of east purple common perilla extractive
CN113398222A (en) * 2020-03-17 2021-09-17 云南中医药大学 National medicine composition for exorcising evil spirits and preventing epidemic

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116392518A (en) * 2023-04-19 2023-07-07 昆明理工大学 Application of perilla and extract thereof in preparing medicine for treating chronic pressure brain lesions
CN116392518B (en) * 2023-04-19 2024-03-15 昆明理工大学 Application of perilla and extract thereof in preparing medicine for treating chronic pressure brain lesions

Also Published As

Publication number Publication date
CN114949034B (en) 2023-11-07

Similar Documents

Publication Publication Date Title
CN104844723A (en) Preparation method and application of dendrobium officinale extract
CN115531473B (en) Compound essential oil for treating skin diseases and pharmaceutical composition containing same
CN111870568B (en) Anti-allergy itching-relieving plant composition and preparation method and application thereof
CN114949034B (en) Application of perilla extract in preparing medicine for preventing and treating acute pneumonia
CN105617316A (en) Antibacterial disinfectant for clinical laboratory and preparation method
CN104208294B (en) It is a kind of to be used to treat Chinese medicine composition of piglet yellow-white dysentery and preparation method thereof
KR20130121324A (en) Composition for preventing and treating hangover comprising extracts of leaves of perilla frutescens britton var
CN108619165A (en) Application of the Polysaccharide from Scutellaria Baicalensis in preparing veterinary drug
CN108542926A (en) The preparation method and pharmaceutical composition of a kind of purslane extract and its application
CN110882332B (en) Antibacterial, anti-inflammatory and antiallergic nasal traditional Chinese medicine composition and preparation method and application thereof
CN108969580B (en) Preparation method and application of blue cloth total tannin
CN112920927A (en) Traditional Chinese medicine compound vinegar composition and preparation method thereof
Andilala Safraian (2023). The Effectiveness of Kersen Leaf Extracts (Muntingia calabura L.) on Cutting Wounds Infected with Sthapylococcus Aureus Bacteria
KR20200040427A (en) Method for producing medicine of skin diseases
KR102223717B1 (en) The crude drugs composition of decreation or treating chronic obstructive pulmonary disease, phamacopuncture injection of oriental medicinie containing the same and Injection containing the same
CN113069466B (en) Application of purslane polysaccharide in preparation of acute lung injury resistant medicine
CN116492369B (en) Traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects and application thereof
CN115844933B (en) Application of clindamycin total flavone in preparing heart failure resisting medicine
CN103083376B (en) Can be used for the Rhizoma Ligustici extract for the treatment of rhinitis
KR20120060053A (en) Composition for the anti-inflammatory and analgesia comprising extract of propolis and Sophora japonica complex containing flavonoid
CN106387581A (en) Preparation technology of chicoric acid enriched fresh echinacea purpurea squeezed juice
CN117100819A (en) Application of red rice seed coat active extract in preparation of medicine for preventing and treating acute alcoholic liver disease
CN1961911A (en) Lyophilized powder injection containing red sage root and dalbergia wood volatile oil and preparation method thereof
CN118045115A (en) Application of QIAGUOTA extract in preparing medicine for preventing and treating Alzheimer disease
CN104523740A (en) Method for preparing selaginella doederleinii hieron polysaccharide extract product and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant