Application of compound fresh bamboo juice in young group cough model

Abstract

The invention discloses application of compound fresh bamboo juice in a young group cough model, and relates to the field of medicines.The application comprises the following steps that S1, a cough model is constructed by adopting young beagles, and whether the cough model is successfully constructed or not is verified through behavioral observation, hematological analysis, CT examination and alveolar lavage analysis of the young beagles; and S2, evaluating the efficacy and safety of the successfully constructed young beagle by using a compound fresh bamboo juice. According to the invention, a technical system for evaluating the efficacy of the compound fresh bamboo juice in the cough model of the young beagle is established by taking the young beagle as a research object. In a puppy beagle model, the compound fresh bamboo juice shows good safety, and the medicine remarkably improves target symptoms. The results provide powerful support for the potential curative effect of the compound fresh bamboo juice in children.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and specifically to the application of Compound Fresh Bamboo Extract Liquid in a cough model for the juvenile population. Background Art

[0002] At present, respiratory diseases are highly prevalent in China, and the market share of traditional Chinese patent medicines is large. As a major cough-relieving and phlegm-dissipating drug, Compound Fresh Bamboo Extract Liquid has strong market demand. The following studies have been conducted: (1) Observe and analyze the effect of atomizing inhalation of the traditional Chinese medicine preparation Compound Fresh Bamboo Extract Liquid in patients with post-infectious cough. (2) Verify the efficacy of Compound Fresh Bamboo Extract Liquid in aspects such as anti-inflammatory, expectorant, and cough-suppressing effects. (3) Explore the acute and long-term toxic reactions of Compound Fresh Bamboo Extract Liquid in mice and rats.

[0003] In clinical medicine, there are significant differences between adults and children in terms of physiological structure, immune function, and disease susceptibility, which lead to differences in the cough mechanism, clinical manifestations, and treatment strategies between the two. The causes of adult cough are complex and diverse, including but not limited to chronic respiratory diseases, gastroesophageal reflux, cardiovascular diseases, etc., and environmental factors such as smoking and air pollution are also non-negligible factors. In contrast, childhood cough is more commonly seen in viral infections, allergic reactions, and airway developmental abnormalities, with relatively single causes but rapid disease progression.

[0004] Therefore, the existing technologies mainly have the following problems: (1) The components of Compound Fresh Bamboo Extract Liquid are complex, lacking research support for the efficacy values and effective mechanisms of action in special groups (juveniles). (2) The drug lacks research on the safe use of special groups (juveniles), and there are certain risks in clinical application. It is difficult for Western medicine clinicians to accurately understand the indications and medication methods of Compound Fresh Bamboo Extract Liquid, which affects its clinical application effect.

[0005] Currently, the drug safety and efficacy of Compound Fresh Bamboo Extract Liquid in special populations (children) or pets (dogs) have not been clearly defined.

[0006] With the development of modern technologies, new technical means are constantly being introduced into the efficacy research of Compound Fresh Bamboo Extract Liquid. Currently, there are design schemes for precise dosing of animal models and treatment studies of Compound Fresh Bamboo Extract Liquid on animal models. Summary of the Invention

[0007] The purpose of the present invention is to solve at least one of the technical problems existing in the prior art, and provide the application of Compound Fresh Bamboo Extract Liquid in a cough model for the juvenile population.

[0008] The technical solution of the present invention is as follows:

[0009] The application of Compound Fresh Bamboo Extract Liquid in a cough model for the juvenile population includes the following steps:

[0010] S1: Establish a cough model using juvenile beagle dogs. Verify whether the cough model is successfully established through behavioral observation, hematological analysis, bronchoalveolar lavage analysis, and CT examination of juvenile beagle dogs.

[0011] S2: Use Compound Fresh Bamboo Extract to evaluate the efficacy and safety of the successfully established cough model.

[0012] Further, in step S1, the behavioral observation includes animal coughing and wheezing, airway secretions, respiratory rate, cyanosis, and presence or absence of dyspnea.

[0013] The hematological analysis includes differential count of venous blood white blood cells and the expression of immunoglobulin E and immunoglobulin G in serum.

[0014] The bronchoalveolar lavage analysis includes differential count of white blood cells in bronchoalveolar lavage fluid.

[0015] Further, in step S1, the juvenile beagle dogs are male, 3 - 4 months old, and weigh 3 - 4 kg.

[0016] Further, in step S2, the method for evaluating efficacy and safety is specifically as follows:

[0017] Adaptively raise the juvenile beagle dogs for 1 week, start modeling, administer 0.4 ml of 4 mmol / L capsaicin suspension into the bronchus every day. After modeling, observe and record the number of coughs within 10 minutes. Administer the corresponding volume of Compound Fresh Bamboo Extract or noscapine tablets by gavage 30 minutes after modeling. Conduct routine physical examinations, hematological examinations, and CT examinations on day 0, 7, and 15 after modeling. Perform cell counting of the right middle lobe alveolar lavage at 15 days after modeling, and take pathological examinations of the other lungs to evaluate the inhibitory effects of the drug on cough and inflammatory factors.

[0018] Adaptively raise the juvenile beagle dogs for 1 week. After single - dose gavage administration, observe the effects of the drug on the cardiovascular and respiratory system functions of the juvenile beagle dogs, as well as the changes in key indicators such as blood routine, blood biochemistry, and blood gas, so as to evaluate the reliability of the safety pharmacology and toxicology of this drug in clinical application.

[0019] Further, the cough efficacy indicators include behavioral observation, hematological analysis, bronchoalveolar lavage analysis, and CT examination.

[0020] Further, the safety indicators include overall observation, clinical examination, cardiovascular system examination, and respiratory system examination.

[0021] Further, Compound Fresh Bamboo Extract can relieve cough in the juvenile population cough model and has no negative impact on the hematopoietic system.

[0022] The beneficial effects of the present invention are as follows: Using juvenile Beagle dogs as the research object, a pharmacodynamic evaluation technical system of compound fresh bamboo juice liquid in the cough model of juvenile Beagle dogs was established.

[0023] In the Beagle puppy model, compound fresh bamboo juice liquid showed good safety, and the drug significantly improved the target symptoms. These results provide strong support for the potential efficacy of compound fresh bamboo juice liquid in children. Description of the Drawings

[0024] Figure 1 It is the statistical result of the cough times of Beagle dogs after administration in each group of the present invention;

[0025] Figure 2 It is the blood routine test result of Beagle dogs in each group of the present invention;

[0026] Figure 3 It is the imaging result of the lung CT examination of Beagle dogs of the present invention;

[0027] Figure 4 It is the examination result of the alveolar lavage fluid of the Beagle dog diagram of the present invention;

[0028] Figure 5 It is the H&E and PSR staining results of Beagle dogs of the present invention;

[0029] Figure 6 It is the clinical cough performance of Beagle dogs in each treatment group of the present invention;

[0030] Figure 7 It is the hematological analysis result of Beagle dogs in each treatment group of the present invention;

[0031] Figure 8 It is the alveolar lavage result of Beagle dogs in each treatment group of the present invention;

[0032] Figure 9 It is the analysis result of the Brixia scoring system of the CT examination of Beagle dogs in each treatment group of the present invention;

[0033] Figure 10 It is the flow chart of the pharmacodynamic study on the cough effect of the present invention on juvenile Beagle dogs;

[0034] Figure 11 It is the flow chart of the potential impact of the present invention on the cardiovascular and respiratory systems of juvenile Beagle dogs. Detailed Embodiments

[0035] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used that do not specify the manufacturer are all conventional products that can be obtained commercially.

[0036] This section will describe in detail the specific embodiments of the present invention. The preferred embodiments of the present invention are shown in the accompanying drawings. The purpose of the accompanying drawings is to supplement the description of the text part of the specification with graphics, so that people can intuitively and vividly understand each technical feature and the overall technical solution of the present invention, but it cannot be understood as a limitation on the scope of protection of the present invention.

[0037] The compound fresh bamboo sap liquid used in this embodiment is produced and provided by Jiangxi Jiuhua Pharmaceutical Co., Ltd. (approval number: National Medicine Standard Z36020947; drug production batch number: 07230204), the drug type is a Chinese patent medicine, the prescription type is an over-the-counter drug OTC, the main ingredients of the drug are: fresh bamboo sap, houttuynia cordata, loquat leaf, platycodon, raw pinellia, ginger, peppermint oil (or peppermint oil), and the auxiliary materials are sucrose, purified water, and sodium benzoate.

[0038] 1. Construct a cough model for young Beagle dogs and improve its cough model evaluation indicators

[0039] In animal experiments, different animals have their own advantages and disadvantages in the study of cough. For example, guinea pigs are sensitive to chemical or mechanical stimulation and are often used in the study of bronchodilation. They are the preferred animals for screening antitussive drugs, but their cough reflex is quite different from that of humans, and the experimental reliability is poor. The cough reflex of rats and mice is even more different from that of humans, and the experimental reliability is even lower. Cats rarely cough under physiological conditions, while the cough reflex mechanism of dogs is closer to that of humans, which is particularly suitable for observing the duration of the antitussive effect of drugs. In order to better evaluate the therapeutic efficacy of compound fresh bamboo sap in cough, we chose dogs as a species that is closer to human cough, and beagles are standard experimental dogs, so in this experimental plan, beagles were used as research subjects.

[0040] Modeling: The subjects were male beagles, 3-4 months old, 3-4 kg. After anesthesia, nasal cannula was used for drug administration, with a depth of 15 cm from the corner of the animal's mouth. The control group (Sham) was given 0.4 mL of normal saline, and the model groups model A (capsaicin suspension 1 mmoL / L), model B (capsaicin suspension 2 mmoL / L), and model C (capsaicin suspension 4 mmoL / L) were given 0.4 mL respectively. The tube was removed immediately after drug administration. The respiratory status of the animals was closely monitored during the observation process. The results are as follows:

[0041] Figure 1 andFigure 2 It can be seen that during the model establishment process, there were no obvious changes in the animal red blood cells, indicating that the model establishment process had no impact on the animal hematopoietic system. Compared with the control group, after 7 days of model establishment, the white blood cells in the model group all increased, mainly due to the increase in neutrophils.

[0042] Figure 3 From the imaging, on the 7th day, high-density shadows appeared in the lungs of the dogs in model C group, with blurred boundaries and thickened internal textures, suggesting inflammatory changes; on the 15th day, large-area high-density shadows appeared in the middle and lower lobes of the lungs of the dogs in model C group, with blurred boundaries and thickened internal textures, suggesting inflammatory changes. No clear necrosis or cavity formation was seen in the lesions, but local bronchial wall thickening could be observed. The surrounding lung tissue was compressed, and the lung textures in some areas were thickened and disordered. It was considered that model establishment could lead to inflammatory changes in the animal lungs and bronchial thickening.

[0043] Figure 4 and Figure 5 It can be seen that the neutrophils in the bronchoalveolar lavage fluid of the model group increased sharply. As the dose of the model establishment drug increased, the alveolar tissue structure gradually became disordered and the alveolar wall thickened. There was a certain degree of pulmonary fibrosis, and this change often occurred in the late stage of the inflammatory reaction. The inflammatory infiltration in the bronchi and trachea in the lung tissue increased, indicating that a strong inflammatory reaction was occurring in the bronchi. The above results showed that this method could successfully establish a cough model.

[0044] 2. Therapeutic Efficacy and Safety of Compound Fresh Bamboo Liquid in the Cough Model of Juvenile Beagle Dogs

[0045] 2.1 Pharmacodynamic Study of Compound Fresh Bamboo Liquid in a Special Group (Juvenile)

[0046] Taking juvenile beagle dogs as the research objects, by establishing a cough animal model, it was divided into a low-dose group (0.57 ml / kg), a medium-dose group (1.14 ml / kg), and a high-dose group (2.28 ml / kg) to verify the drug efficacy of compound fresh bamboo liquid in juvenile beagle dogs and explore the dose-effect relationship at the same time.

[0047] Experimental subjects: Beagle dogs, male, 3 - 4 months old, 3 - 4 kg, 6 in each of the control group, model group, low-dose group, medium-dose group, high-dose group, and positive drug group, a total of 36;

[0048] Experimental procedure: Refer to Figure 10;Modeling method: Young beagle dogs were fed adaptively for 1 week. Subsequently, the modeling began. Approximately 3 ml of propofol was injected intravenously to briefly anesthetize them. After anesthesia, tracheal drug administration was performed via a nasal catheter. The catheter was inserted 15 cm from the corner of the animal's mouth, and then 0.4 ml of 4 mmol / L capsaicin was instilled. After the modeling was completed, the coughing situation of the animals was observed, and the number of coughs of the animals within 10 minutes was counted. Experimental drug administration method: An F20 gastric tube was selected. The animal's mouth was opened with an oral speculum, and the catheter was inserted into the stomach through the pharynx and esophagus. The corresponding dose of the drug was injected with a syringe, and the catheter was then rinsed with physiological saline with a volume twice that of the drug. Finally, the tube was pinched and smoothly withdrawn.

[0049] Experimental detection indicators:

[0050] a. Behavioral observation: Clinical observation: The time after drug administration was T0, with 1 min as a time point, and the number of coughs within 10 min was objectively recorded. The results are as follows:

[0051] From Figure 6 the clinical cough manifestations, it can be seen that a cough model of young beagle dogs was constructed and treated with drugs. During the treatment process, there were significant differences in the number of coughs between the drug administration group and the model group. The drug administration group had a certain therapeutic effect, and the high-dose group was more obvious.

[0052] b. Routine physical examination: Daily physical examinations were performed on days 0, 7, and 15, including body weight, etc.

[0053] c. Blood routine examination: Blood routine examinations were performed on days 0, 7, and 15 respectively, mainly observing the changes in red blood cells and white blood cells.

[0054] d. CT examination: CT examination was performed on the animals on day 0 for pre-treatment control. On day 7, 1 CT was performed on each of the different dose groups in the model group. The animals selected were those that could show severe coughs after daily drug administration and whose daily results were relatively stable. Terminal examination was performed on day 15. CT examination content: Whether the bronchial wall was thickened, dilated or stenosed, and whether there was infection in the lungs. Brixia scores were performed on all CT results;

[0055] e. Pathological examination: HE staining and PSR staining were used to analyze the pulmonary inflammatory reaction and bronchial mucus secretion. ① 5 - 8 cm from the larynx of the main trachea, ② the upper left lobe of the lung, ③ the lower left lobe of the lung, ④ the upper right lobe of the lung, ⑤ the lower right lobe of the lung, ⑥ the left main bronchus, ⑦ the right main bronchus.

[0056] f. Bronchoalveolar lavage fluid examination: The cell count of the alveolar lavage fluid was measured.

[0057] 2.2. Animal drug safety evaluation of Compound Fresh Bamboo Liquid for special population (children)

[0058] Taking juvenile beagle dogs as the research object, observing the effects of oral administration on the cardiovascular and respiratory systems of conscious Beagle dogs, and providing reference for clinical trials. The specific experiment is as follows:

[0059] Experimental subjects: Beagle dogs, male, 3 - 4 months old, 3 - 4 kg, 24 dogs;

[0060] The experimental procedure refers to Figure 11 , specifically, the juvenile Beagle dogs were adaptively raised for 1 week, and the effects of single - dose gavage on the cardiovascular and respiratory system functions of Beagle dogs, as well as the changes in key indicators such as blood routine, blood biochemistry, and blood gas, were observed.

[0061] Experimental detection indicators:

[0062] a. Overall observation: Observe the physical state, movement state, eating, drinking, urination, defecation, etc. of Beagle dogs before and after drug administration.

[0063] b. Clinical tests: Indicators such as hematology, coagulation, and serum biochemistry.

[0064] c. Cardiovascular system examination: Measure the changes in blood pressure (including systolic blood pressure, diastolic blood pressure, and mean blood pressure, etc.), electrocardiogram (including QT interval, PR interval, QRS wave, etc.), and heart rate, etc. before and after drug administration.

[0065] d. Respiratory system examination: Measure the changes in various respiratory function indicators of animals before and after drug administration, such as respiratory rate, tidal volume, respiratory depth, etc.

[0066] Dogs are particularly suitable for observing the duration of the antitussive effect because the cough reflex mechanism of dogs is somewhat similar to that of humans, and their cough duration is relatively long, which is suitable for evaluating the long - term antitussive effect of drugs. The physiological structure of puppies is similar to that of human infants, so the cough symptoms they show in the laboratory can be used as a reference for cough in human infants.

[0067] From Figures 7 - 9 the hematological analysis, bronchoalveolar lavage analysis, and CT examination to further evaluate the therapeutic effect of the drug. The results show that the drug has no negative impact on the hematopoietic system, and there is an obvious advantage in the high - dose group for controlling inflammation. The results show that the inflammatory cells in the puppies in the three drug - administration groups are significantly decreased compared with the control group, and the lung tissue inflammation in the high - dose group is the mildest.

[0068] 3. Taking juvenile beagle dogs as the research object, exploring the application prospect of Compound Fresh Bamboo Extract in the treatment of cough and lung inflammation in pet dogs. The above - mentioned experiments have proved that Compound Fresh Bamboo Extract can be used in the treatment of cough and lung inflammation in pet dogs.

[0069] In summary, the present invention has the following content:

[0070] (1) An accurate dosage regimen was designed based on the weight and age of juvenile beagle dogs.

[0071] (2) A cough model of juvenile Beagle dogs was constructed, and its cough model evaluation indicators were improved.

[0072] (3) The compound fresh bamboo juice was used to treat juvenile beagle dogs.

[0073] (4) Specific methods for evaluating the efficacy and safety of compound fresh bamboo juice were established.

[0074] Compared with existing studies, the present invention has the following advantages:

[0075] (1) The construction method and evaluation index system of the cough model of juvenile beagle dogs.

[0076] (2) The therapeutic effect of the drug in a special group (juvenile) was evaluated from aspects such as behavioral observation, hematological analysis, bronchoalveolar lavage analysis, and CT examination.

[0077] (3) The safety evaluation of the drug in a special group (juvenile) was carried out from multiple dimensions such as overall observation, clinical examination, cardiovascular system examination, and respiratory system examination.

[0078] (4) The present invention also proves that compound fresh bamboo juice can be used for the treatment of cough and lung inflammation in pet dogs.

[0079] On the premise of no conflict, those skilled in the art can freely combine and superimpose the above additional technical features.

[0080] The above embodiments only represent the preferred embodiments of the present invention, and the description is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, other corresponding changes and deformations can also be made according to the technical solutions and concepts described above, and all these changes and deformations should fall within the protection scope of the claims of the present invention.

Claims

1. The application of Compound Fresh Bamboo Extract Liquid in a cough model for the juvenile population, characterized in that, It includes the following steps: S1: Establish a cough model using juvenile beagle dogs. Verify whether the cough model is successfully established through behavioral observation, hematological analysis, bronchoalveolar lavage analysis, and CT examination of juvenile beagle dogs; S2: Use Compound Fresh Bamboo Lek to evaluate the efficacy and safety of the successfully established cough model.

2. Use of the compound fresh bamboo juice liquid according to claim 1 in a cough model of the juvenile population, characterized in that, In step S1, the behavioral observation includes animal coughing and wheezing, airway secretions, respiratory rate, cyanosis, and presence or absence of dyspnea; The hematological analysis includes differential count of venous blood white blood cells and the expression of immunoglobulin E and immunoglobulin G in serum; The bronchoalveolar lavage analysis includes differential count of white blood cells in bronchoalveolar lavage fluid.

3. The application of the compound fresh bamboo juice liquid according to claim 1 in the cough model of the juvenile group, characterized in that, In step S1, the juvenile beagle dogs are male, 3 - 4 months old, and weigh 3 - 4 kg.

4. Use of the compound fresh bamboo juice liquid according to claim 1 in a cough model for the juvenile population, characterized in that, In step S2, the specific method for evaluating efficacy and safety is as follows: Adaptively raise juvenile beagle dogs for 1 week, and observe the effects on the cardiovascular and respiratory system functions of beagle dogs after single - dose gavage administration of Compound Fresh Bamboo Lek, as well as the changes in key indicators of blood routine, blood biochemistry, and blood gas; Adaptively raise juvenile beagle dogs for 1 week, start modeling, administer Compound Fresh Bamboo Lek, and observe the experimental examination of safety indicators of juvenile beagle dogs after administration.

5. Use of the compound fresh bamboo juice liquid according to claim 4 in a cough model for the juvenile group, characterized in that, The cough efficacy indicators include behavioral observation, hematological analysis, bronchoalveolar lavage analysis, and CT examination.

6. Use of the compound fresh bamboo juice liquid according to claim 4 in a cough model of the juvenile population, characterized in that, The safety indicators include overall observation, clinical examination, cardiovascular system examination, and respiratory system examination.

7. Use of the compound fresh bamboo juice liquid according to claim 1 in a cough model for the juvenile population, characterized in that, Compound Fresh Bamboo Lek can relieve cough in the juvenile population cough model and has no negative impact on the hematopoietic system.

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