Physalis alkekengi extract, preparation method thereof and application of physalis alkekengi extract in preparation of medicine for treating lung injury caused by cigarette smoke

Abstract

The invention relates to a physalis alkekengi extract and a preparation method and application thereof in preparation of medicine for treating lung injury caused by cigarette smoke, and belongs to the field of medicine. The method comprises the steps that fresh physalis alkekengi fruits are added into pure water and continuously concentrated in the water decoction process to obtain pasty fruit pulp, the preliminarily concentrated pasty fruit pulp is put into a freeze dryer to be freeze-dried, and a freeze-dried product is obtained; and collecting to obtain a dried product, and grinding the solid physalis alkekengi into fine powder by using a mortar. Tests prove that the physalis alkekengi extract has an anti-inflammatory effect in non-infectious lung tissue injury, and the effect is excellent. After the physalis alkekengi extract is given, the Bax mRNA expression level of the mouse lung tissue is reduced, and the Bcl2mRNA level is increased, which indicates that the cell apoptosis process of the lung tissue is slowed down, and the lung tissue cell injury is compensated without promoting the formation of cell apoptosis. The physalis alkekengi extract has good anti-oxidation and anti-inflammatory effects and can slow down the apoptosis speed of lung tissue cells and improve lung tissue damage caused by cigarette smoke.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and particularly relates to a Physalis alkekengi extract and a preparation method thereof, and use of the extract in preparing a medicine for treating lung damage caused by cigarette smoke. Background Art

[0002] Lung injury is a kind of substantial organ injury that is extremely harmful and difficult to be detected by people. It is one of the most common diseases with a high incidence rate. Lung injury is easy to occur, difficult to treat, and most of them have a poor prognosis. Severe cases can also lead to death. Smoking is one of the important causes of lung injury. Long-term exposure to cigarette smoke and inhalation of irritating gases and particulate matter can easily cause oxidative stress and induce inflammatory effects. The free radicals produced by smoking inactivate α1-antiproteinase in the lungs and cannot inhibit the elastase released by the aggregation and activation of neutrophils and macrophages produced by smoking in the lungs. This may be an important reason for lung injury caused by smoking. Due to the special structure of the lungs, the inflammatory response caused by oxidative stress is not easy to eliminate, and may even progress to chronic pneumonia and lung tissue fibrosis, eventually leading to COPD, and patients experience dyspnea. Under the premise that the risk of cigarette smoke is an acceptable risk, it is crucial to prevent or delay the damage caused by cigarette smoke, which can not only delay CPOD, but also reduce the degree of CPOD and reduce the burden on patients.

[0003] At present, the treatment options for lung injury caused by cigarette smoke are still relatively limited, and hormone drugs are often widely used as the first choice. However, high-dose and frequent use of hormone drugs can lead to a series of adverse reactions, such as immunosuppression, osteoporosis, and hyperglycemia. Therefore, finding drugs that can effectively alleviate lung injury caused by cigarette smoke has become a hot area of ​​research. In recent years, it has been found in the pharmacology research of traditional Chinese medicine that a variety of traditional Chinese medicines and their extracts can effectively improve lung injury, and have the advantages of few side effects, multiple targets, and definite efficacy.

[0004] Physalis, also known as brocade lantern, red mushroom girl, hanging gold lantern, Goli, lantern grass, lantern fruit, roselle, bubble grass, ghost lantern, etc., is called mushroom and girl in the north. The earliest record began in "Shennong's Herbal Classic". "Compendium of Materia Medica" also records that brocade lantern can remove dampness and heat, clear the lungs and treat cough, remove dampness and resolve phlegm, and treat carbuncle. The whole plant and fruit of brocade lantern can be used as medicine, which has the effects of appetizing, aiding digestion, replenishing blood, lowering blood pressure, and refreshing. It also has a certain effect on aplastic anemia. Physalis is rich in active substances such as bitter physalis, physalis sterol A, physalis sterol B, 3-sitosterol, and cholesterol. These substances have good anti-inflammatory and antioxidant effects. Studies have found that Physalis has a good effect on asthma in mice. Ascorbic acid and flavonoids in Physalis are exogenous antioxidants. After entering the body, they can significantly improve the body's ability to scavenge free radicals. Exogenous supplementation of antioxidants can improve the body's resistance to toxic chemicals such as smoke. Therefore, it is speculated that Physalis can have an intervention effect on lung damage caused by smoke. However, there are few studies on Physalis against lung damage caused by smoke. Summary of the invention

[0005] The purpose of the present invention is to provide a Physalis extract and a preparation method thereof and use thereof in preparing a drug for treating lung injury caused by cigarette smoke. The present invention verifies the intervention effect of the Physalis extract on lung injury caused by cigarette smoke in mice by comparing the degree of lung injury caused by cigarettes on mice fed with different concentrations of Physalis extract, and provides an experimental basis for further studying the pharmacological effects of Physalis.

[0006] The technical solution adopted by the present invention to solve the technical problem is as follows:

[0007] The present invention provides a method for preparing a Physalis alkekengi extract, which specifically comprises the following steps:

[0008] Fresh physalis berries are added into pure water, and the berries are continuously concentrated by decoction to obtain a paste-like pulp. The initially concentrated paste-like pulp is placed into a freeze dryer for freeze-drying, and the dried product is collected. The solid physalis berries are then ground into fine powder using a mortar.

[0009] As a preferred embodiment, the usage ratio of the fresh physalis to pure water is in the range of 0.5 to 0.6 g / ml.

[0010] As a preferred embodiment, in the water decoction method, the water decoction times range from 1 to 2 times, the total water decoction time ranges from 3 to 4 hours, and the water decoction temperature ranges from 90 to 100°C.

[0011] As a preferred embodiment, the freeze-drying conditions are temperature: -50 to -40°C, pressure: 1 to 10 Pa.

[0012] As a preferred embodiment, the mesh size of the fine powder is in the range of 60 to 100 meshes.

[0013] The Physalis alkekengi extract is prepared by the method for preparing the Physalis alkekengi extract of the present invention.

[0014] The invention discloses use of the Physalis alkekengi extract prepared by the method for preparing the Physalis alkekengi extract in preparing a medicine for treating lung damage caused by cigarette smoke.

[0015] The beneficial effects of the present invention are:

[0016] The extract of Physalis prepared by the present invention is rich in flavonoids, B vitamins, vitamin C and other ingredients. Flavonoids and vitamin C are known and very effective exogenous antioxidants. Supplementing flavonoids and vitamin C can effectively improve oxidative damage in the body. After administering the extract of Physalis, the alveolar damage of mice was alleviated, the inflammatory response of the lungs was weakened, the SOD level in the lung tissue was significantly increased, and the MDA level was significantly reduced, indicating that the extract of Physalis can remove free radicals in the tissue, replenish antioxidant substances in the body, and improve the activity of antioxidant substances, thereby exerting an antioxidant effect. In addition, after administering Physalis after cigarette smoke modeling, the expression level of inflammatory factors in the lung tissue and alveolar lavage fluid of mice was significantly reduced, indicating that the extract of Physalis has an anti-inflammatory effect in non-infectious lung tissue damage, and the effect is excellent. At the same time, after administering the extract of Physalis, the expression level of Bax mRNA in the lung tissue of mice was downregulated, and the level of Bcl2 mRNA was upregulated, indicating that the apoptosis process of the lung tissue was slowed down, and it was not necessary to compensate for the damage of the lung tissue cells by promoting the formation of apoptosis.

[0017] Therefore, the Physalis alkekengi extract prepared by the present invention has good antioxidant and anti-inflammatory effects, and can also slow down the apoptosis rate of lung tissue cells and improve lung tissue damage caused by cigarette smoke. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results of lung tissue pathology of mice in each group (HE, 20×).

[0019] Figure 2 The figure shows the effect of Physalis on the gene level of lung tissue in each group of mice. In the figure, compared with the control group, #p<0.05; compared with the model group, &p<0.05; compared with the low-dose group of Physalis, νp<0.05; compared with the medium-dose group of Physalis, *p<0.05. DETAILED DESCRIPTION

[0020] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0021] 1 Materials and methods

[0022] 1.1 Reagents and instruments

[0023] 4% paraformaldehyde, hematoxylin and eosin staining solution (Beijing Solebow Technology Co., Ltd.); MDA, SOD, NO kits (Nanjing Jiancheng Engineering Co., Ltd.); IL-1β, IL-6, TNF-α, IL-1, IL-12 ELISA kits (Jingmei Bioengineering Co., Ltd.); Physalis alkekengi (commercially available); Cigarettes: a certain brand of commercially available filter cigarettes (flue-cured tobacco type, tar content 11 mg, smoke nicotine content 0.7 mg).

[0024] Microplate reader (Bioteck); desktop high-speed refrigerated centrifuge (Thermo); electron microscope (Olympus); tissue homogenizer (IKA).

[0025] 1.2 Experimental animals

[0026] Thirty male Kunming SPF mice, weighing 18–22 g, were purchased from Changchun Yisi Experimental Animal Technology Co., Ltd., license number: SCXK (Ji) 2018-0007, animal number: 01021690785926811, and rearing conditions: temperature 18–22 °C, humidity 45–65 °C, and 12 h light–dark cycle.

[0027] 1.3 Preparation of Physalis alkekengi extract

[0028] Add appropriate amount of pure water to fresh physalis berries, wherein the ratio of fresh physalis berries to pure water is 0.55 g / ml, and continuously concentrate through a water decoction process to obtain a paste-like pulp, wherein in the water decoction method, the water decoction times range is 2 times, the total water decoction time range is 3.5 h, and the water decoction temperature range is 95°C; put the initially concentrated paste-like pulp into a freeze dryer for freeze drying, wherein the freeze drying conditions are temperature: -50 to -

[0029] 40℃, pressure: 1~10pa; collect the dried product, and then grind the solid acid pulp into fine powder with a mesh size of 60~100 mesh using a mortar.

[0030] 1.4 Animal grouping and drug administration

[0031] Mice were randomly divided into control group, model group, low-dose, medium-dose and high-dose groups of Physalis according to body weight. Except for the control group, the other groups were placed in a closed container and passively smoked using an atomizer inhaler to poison the mice. For each group of 6 mice, a cigarette was soaked in 30mL of sterile water for 2 hours, and then the sterile water soaked in cigarette tobacco was injected into the atomizer generator. Each atomization was 30 minutes, twice a day, and the modeling time was 8 weeks. The low-dose, medium-dose and high-dose groups of Physalis were gavaged with 200mg / kg, 1000mg / kg and 2000mg / kg of Physalis solution, respectively, once a day for 21 consecutive days. The mice were killed, and the alveolar lavage fluid and lung tissue were collected for detection.

[0032] 1.5 Determination of TNF-α, IL-1, IL-6, IL-12, IL-1β, MDA, NO and SOD contents

[0033] Immediately after the mice were killed, the alveolar lavage fluid was quantitatively collected, centrifuged at 3000rpm for 10min, and the supernatant was taken. The contents of IL-1, IL-6 and IL-12 in the alveolar lavage fluid were determined according to the instructions of the ELISA kit. Whole blood was taken, centrifuged at 3000rpm for 10min, and the supernatant was taken. The contents of THF-A, IL-6 and IL-1β in serum were determined according to the instructions of the kit. Lung tissue was taken, and 10% tissue homogenate was made with physiological saline in an ice bath, centrifuged at 4°C and 3000rpm, and the supernatant was taken. The contents of NO, SOD and MDA in lung tissue were determined strictly according to the instructions of the kit.

[0034] The effects of Physalis extract on the levels of TNF-a, IL-6 and IL-1β in the serum of each group of mice are as follows:

[0035] The results in Table 1 show that the levels of TNF-a, IL-6 and IL-1β in the serum of the mice in the model group were significantly higher than those in the control group (p < 0.05), and the levels of IL-6 in the serum of the mice in the low, medium and model groups of Physalis were significantly lower than those in the model group (p < 0.05). The levels of TNF-a in the serum of the mice in each dose group of Physalis were significantly lower than those in the model group (p < 0.05); the levels of TNF-a in the serum of the mice in the high dose group of Physalis were significantly lower than those in the medium dose group (p < 0.05).

[0036] Table 1 Effects of Physalis sylvestris on TNF-a, IL-6 and IL-1β in serum of mice in each group

[0037] Group TNF-a (ng / L) IL-6 (pg / mL) IL-1β (ng / L) Control group 0.56±0.04 0.43±0.05 0.27±0.14 Model Group <![CDATA[0.79±0.08 # ]]> <![CDATA[0.55±0.07 # ]]> <![CDATA[0.41±0.02 # ]]> Physalis low dose group <![CDATA[0.59±0.06 & ]]> <![CDATA[0.52±0.06 # ]]> 0.38±0.02 Physalis medium dose group <![CDATA[0.66±0.11 & ]]> <![CDATA[0.54±0.03 # ]]> 0.36±0.03 Physalis high dose group <![CDATA[0.52±0.03 &* ]]> 0.48±0.03 0.36±0.01

[0038] Compared with the control group, #p<0.05; compared with the model group, &p<0.05.

[0039] The effects of Physalis alba on the levels of IL-1, IL-6 and IL-12 in bronchoalveolar lavage fluid of mice in each group are as follows:

[0040] As shown in Table 2, the levels of IL-1, IL-6 and IL-12 in the bronchoalveolar lavage fluid of mice in the model group were significantly higher than those in the control group (p < 0.05). The IL-1 in the bronchoalveolar lavage fluid of mice in the medium and high dose groups of Physalis was significantly lower than that in the model group and the low dose group (p < 0.05); the IL-1 in the bronchoalveolar lavage fluid of mice in the high dose group of Physalis was significantly lower than that in the medium dose group (p < 0.05); the IL-6 in the bronchoalveolar lavage fluid of mice in each dose group of Physalis was significantly lower than that in the model group (p < 0.05); the IL-12 in the bronchoalveolar lavage fluid of mice in the medium and high dose groups of Physalis was significantly lower than that in the model group (p < 0.05); the IL-12 in the bronchoalveolar lavage fluid of mice in the high dose group of Physalis was significantly lower than that in the low dose group (p < 0.05); with the increase of Physalis concentration, the expression level of inflammatory factors in the bronchoalveolar lavage fluid of mice gradually decreased.

[0041] Table 2 Effects of Physalis sylvestris on IL-1, IL-6 and IL-12 in bronchoalveolar lavage fluid of mice in each group

[0042]

[0043]

[0044] Compared with the control group, #p<0.05; compared with the model group, &p<0.05; compared with the low-dose group of Physalis, νp<0.05; compared with the medium-dose group of Physalis, *p<0.05.

[0045] The effects of Physalis alkekengi on the contents of NO, MDA and SOD in the lung tissues of mice in each group are as follows:

[0046] As shown in Table 3, the MDA content in the lung tissue of mice in the model group was significantly higher than that in the control group, and the SOD content was significantly lower than that in the control group (p < 0.05); the MDA in the lung tissue of mice in the medium and high dose groups of Physalis was significantly lower than that in the model group and the low dose group (p < 0.05); the SOD in the low, medium and high dose groups of Physalis was significantly higher than that in the model group (p < 0.05); the SOD in the medium and high dose groups of Physalis was significantly higher than that in the low dose group (p < 0.05); the SOD in the high dose group of Physalis was significantly higher than that in the medium dose group (p < 0.05); the NO in the low and medium dose groups of Physalis was significantly lower than that in the model group (p < 0.05); the NO in the high dose group of Physalis was significantly higher than that in the low and medium dose groups (p < 0.05).

[0047] Table 3 Effects of Physalis alkekengi on NO, MDA and SOD in lung tissues of mice in each group

[0048] Group NO(μmol / g) MDA (μmol / g) SOD(U / mg) Control group 16.12±0.33 9.09±2.05 122.86±5.31 Model Group 28.03±3.34# 16.10±0.66# 16.23±5.31# Physalis low dose group 14.69±0.12& 15.06±0.57# 91.56±2.01#& Physalis medium dose group 14.55±0.22& 12.03±1.57#&ν 105.47±4.02#&ν Physalis high dose group 18.90±11.55&ν* 11.08±0.01&ν 117.06±2.01&ν*

[0049] Compared with the control group, #p<0.05; compared with the model group, &p<0.05; compared with the low-dose group of Physalis, νp<0.05; compared with the medium-dose group of Physalis, *p<0.05.

[0050] 1.6 qRT-PCR determination of gene levels in mouse lung tissue

[0051] Skin tissue samples were cut and total RNA was extracted by adding Trizol reagent. The purity and concentration of RNA were evaluated using NanoDrop 2000 UV-Vis spectrophotometer. cDNA was synthesized using StartScript First-Strand cDNA Synthesis Kit (GenStar, Co.). qRT-PCR detection (7500Fast, ABI) was performed after adding primers and RealStar Mix (GenStar, Co.).Interleukin 1β (IL-1β) upstream primer: 5'-GCAACTGTTCCTGAACTCAACT-3', downstream primer: 5'-ATC TTTTGGGGTCCGTCAACT-3'; interleukin 6 (IL-6) upstream primer: 5'-AGTTGCCTTCTTGGGACTGA-3', downstream primer: 5'-ACTGGTCTGTTGTGGGT GGT-3'; interleukin -17A (IL-17A) upstream primer: 5'-TTTAACTCC CTTGGCGCAAAA-3', downstream primer: 5'-CTTTCCCTCCGCATTGACAC-3'; tumor necrosis factor α (TNF-α) upstream primer: 5'-TTCCGTCCCTCTC ATACACTG-3', downstream primer: 5'-AGACACCGCCTGGAGTTCT-3'; chemokines (CXCL1) upstream primer: 5'-CTGGGATTCACCTCAAGAACATC-3', downstream primer: 5'-CAGGGTCAAGGCAAGCCTC-3'; interferon gamma (IF N-γ) upstream primer: 5'-ATGAACGCTACACACTGCATC-3', downstream primer: 5'-CCAT CCTTTTGCCAGTTCCTC-3'; B-cell lymphoma-2 (bcl-2) upstream primer: 5'-GACTTTGCAGAGATGTCCAG-3', downstream primer: 5'-TCAGGTACTC AGTCATCCAC-3'; BCL2-associated X protein (BCL2-associated X protein) protein, bax) upstream primer: 5'-TTTCATCCAGGATCGAGCAG-3', downstream primer: 5'-AATCATCCTCTGC AGCTCCA-3'; Activating Transcription Factor 6 (ATF6) upstream primer: 5'-GACTCACCCATCCGAGTTGTG-3', downstream primer: 5'-CTCCCAGTCTT CATCTGGTCC-3' and GAPDH upstream primer: 5'-GGTCCCAGCTTAGGTTCATCA-3', downstream primer: 5'-CCGTTCACACCGACCTTCA-3'.

[0052] The effects of Physalis on the levels of genes such as ATF6, Bax, and Bcl2 in mouse lung tissue are as follows:

[0053] Depend on Figure 2 It can be seen that qRT-PCR was used to analyze the mRNA expression levels of inflammatory factors (IL-6, TNF-α and IL-1β), apoptotic factors (bax, bcl2 and ATF6) and oxidative damage factors (IL-17A, CXCL1 and IFN-γ) in the lung tissues of each group of mice. Compared with the control group, the gene expression levels of mice in the model group were significantly different (p < 0.05). Figure 2 (a) The ATF6 mRNA level in the lung tissue of mice in the medium and high dose groups of Physalis was significantly lower than that in the model group (p < 0.05); the ATF6 mRNA level in the lung tissue of mice in the high dose group of Physalis was significantly lower than that in the low and medium dose groups (p < 0.05); Figure 2 (b), the bax mRNA levels in the lung tissues of mice in each dose group of Physalis were significantly lower than those in the model group (p < 0.05); Figure 2 (c) The bcl2 mRNA level in the lung tissue of mice in the medium and high dose groups of Physalis was significantly higher than that in the model group (p < 0.05); the bcl2 mRNA level in the lung tissue of mice in the high dose group of Physalis was significantly higher than that in the low dose group (p < 0.05); Figure 2 (d) The CXCL1 mRNA level in the lung tissue of mice in each dose group of Physalis was significantly lower than that in the model group (p < 0.05); the CXCL1 mRNA level in the lung tissue of mice in the high dose group of Physalis was significantly lower than that in the low dose group (p < 0.05); Figure 2 (e) The IL-17A mRNA level in the lung tissue of mice in each dose group of Physalis was significantly lower than that in the model group (p < 0.05); the IL-17A mRNA level in the lung tissue of mice in the medium and high dose groups of Physalis was significantly lower than that in the low dose group (p < 0.05); Figure 2 (f) The IFN-γ mRNA levels in the lung tissues of mice in each dose group of Physalis were significantly lower than those in the model group (p < 0.05); the IFN-γ mRNA levels in the lung tissues of mice in the medium and high dose groups of Physalis were significantly lower than those in the low dose group (p < 0.05); the IFN-γ mRNA levels in the lung tissues of mice in the high dose group of Physalis were significantly lower than those in the medium dose group (p < 0.05); Figure 2 (g) The IL-6 mRNA level in the lung tissue of mice in each dose group of Physalis was significantly lower than that in the model group (p < 0.05); the IL-6 mRNA level in the lung tissue of mice in the high dose group of Physalis was significantly lower than that in the low dose group (p < 0.05); Figure 2 (h), the TNF-α mRNA level in the lung tissue of mice in each dose group of Physalis was significantly lower than that in the model group (p<0.05); the TNF-α mRNA level in the lung tissue of mice in the high dose group of Physalis was significantly lower than that in the low dose group (p<0.05); Figure 2 (i) The IL-1β mRNA levels in the lung tissues of mice in each dose group of Physalis alkekengi were significantly lower than those in the model group (p<0.05).

[0054] 1.7 Histomorphological observation

[0055] The lung tissues of mice were embedded, sectioned, and stained with HE, and the morphological and structural changes of the lung tissues were observed under a light microscope.

[0056] Pathological examination results such as Figure 1 As shown in the figure, the lung tissue structure of mice in the control group was intact and clear, and no changes such as thickening, congestion or edema of the alveoli were observed. The alveoli of mice in the model group were damaged, the alveolar walls were thinned, the boundaries were unclear, and there was inflammatory cell infiltration. The lung tissue damage in each dose group was significantly improved, and the squeezing and inflammatory changes were alleviated.

[0057] 1.8 Statistical analysis

[0058] In this invention, SPSS 18.0 software was used for data processing, and all data were expressed as mean ± standard deviation. The means among the groups were compared by one-way analysis of variance, the pairwise comparison between the groups was performed by the least significant difference (LSD) test, and the comparison between the two groups was performed by the t test, with the test level α = 0.05.

[0059] Under physiological conditions, endogenous antioxidants in lung tissue (such as superoxide dismutase, catalase, glutathione peroxidase, etc.) are in dynamic equilibrium with free radicals produced in the body. However, with the increase of damage, aging, malnutrition and other factors, endogenous antioxidants will be lost and their activity will decrease, causing the accumulation of free radicals produced in the body and triggering oxidative stress. When the body is exposed to exogenous chemicals (such as particulate matter, irritating gases), it will also induce oxidative stress in lung tissue. At this time, the intake of exogenous antioxidants is crucial. The intake of exogenous antioxidants will enhance the body's ability to scavenge free radicals, slow down the oxidative stress response of lung tissue, and thus slow down lung tissue damage.

[0060] Malondialdehyde (MDA) is a commonly used lipid peroxide biomarker, which is often used to evaluate the level of oxidative stress; the content of superoxide dismutase (SOD) reflects the antioxidant capacity of tissues. Lung damage is usually accompanied by the consumption of SOD in the tissues, which reduces the tissue's ability to reduce superoxides, further promoting external stimuli to damage lung tissues.

[0061] Alveolar macrophages are a key factor in the pathogenesis of lung tissue injury. Infiltrating alveolar macrophages mainly promote the progression of inflammation in the early stage of lung tissue injury, and show anti-inflammatory effects in the late stage. These phenotypes can be attributed to macrophage phenotypic transformation. Alveolar macrophages can be divided into M1 macrophages that promote inflammation and M2 macrophages that inhibit inflammation. Studies have shown that when lung injury occurs, monocytes in the peripheral blood are chemotactic and reach the lungs, especially the alveolar cavity, and differentiate into M1 macrophages. M1 macrophages will secrete a large number of pro-inflammatory factors such as IL-1β, IL-6, IL-2 and TNF-α, recruit neutrophils into the lung tissue and alveolar cavity, and induce severe inflammatory response.

[0062] IL-17A (interleukin 17A, also known as CTLA8) is a member of the IL-17 receptor family. The protein encoded by this gene is a proinflammatory cytokine produced by activated T cells. Downstream pathways mediated by IL-17A induce the production of inflammatory molecules, chemokines, antimicrobial peptides, and remodeling proteins. The encoded protein has an important impact on host defense, cell trafficking, immune regulation, and tissue repair, and plays a key role in inducing innate immune defense. This cytokine stimulates non-hematopoietic cells and promotes the production of chemokines, thereby recruiting myeloid cells to the site of inflammation. This cytokine also regulates the activity of NF-κB and mitogen-activated protein kinases, can stimulate the expression of IL6 and cyclooxygenase 2 (PTGS2 / COX-2), and enhance the production of nitric oxide (NO). IL-17 can induce macrophages to secrete proinflammatory cytokines such as TNF-a, IL-1β, and IL-6. IL-17 can also stimulate the production of multiple chemokines, including IL-8 / CXCL8, CXCL1, CXCL2, CCL20, CCL2, and CCL7, which recruit granulocytes, macrophages, and lymphocytes to the site of injury, thereby aggravating inflammation.

[0063] Chemokine (CXC motif) ligand 1 (CXCL1) is a member of the CXC chemokine family. It can act as a chemoattractant for a variety of immune cells, especially chemotactic neutrophils to reach the site of injury or infection, and plays an important role in regulating immune and inflammatory responses. CXCL1 will be highly expressed during the inflammatory response, thereby promoting the inflammatory process.

[0064] IFN-γ is a soluble dimeric cytokine, and its medical name is interferon-γ. Its main function in the body is to induce the synthesis of indoleamine 2, 3-dioxygenase, the production of nitric oxide synthase, and promote NO synthesis, and play an immunomodulatory role. When evaluating the progression of airway inflammation such as COPD, pneumonia, and lung injury, the level changes of factors such as IL-6, TNF-α, and IFN-γ are usually evaluated to evaluate the progression, outcome, and efficacy of the disease.

[0065] In the present invention, a mouse lung tissue injury model was established by cigarette smoke inhalation method, and it was observed that cigarette smoke could not only induce oxidative damage, but also cause inflammatory response, leading to increased levels of pro-inflammatory factors in mouse bronchoalveolar lavage fluid and lung tissue, increased levels of oxidative damage markers MDA and NO, but decreased activity of antioxidant enzyme SOD.

[0066] The Physalis used in the present invention is rich in flavonoids, B vitamins, vitamin C and other ingredients. Flavonoids and vitamin C are known and very effective exogenous antioxidants. Supplementing flavonoids and vitamin C can effectively improve oxidative damage in the body. It was observed in the present invention that after administering Physalis, the alveolar damage of mice was alleviated, the inflammatory response of the lungs was weakened, the SOD level in the lung tissue was significantly increased, and the MDA level was significantly reduced, indicating that Physalis can remove free radicals in the tissue, supplement the antioxidant substances in the body, and increase the activity of antioxidant substances, thereby exerting an antioxidant effect. In addition, after administering Physalis after cigarette smoke modeling, the expression level of inflammatory factors in the lung tissue and bronchoalveolar lavage fluid of mice was significantly reduced, indicating that Physalis has an anti-inflammatory effect in non-infectious lung tissue damage, and the effect is excellent.

[0067] Bcl-2 and Bax are regulatory genes of cell apoptosis, and their expression levels directly reflect the level of cell apoptosis. Bcl-2 inhibits cell apoptosis, Bax promotes cell apoptosis, and the balance between the two maintains the normal metabolism of cells. When Bax and Bcl-2 form Bax-Bcl-2 heterodimers, cell apoptosis is inhibited, while when Bax and Bax form homodimers Bax-Bax, cell apoptosis is promoted. The experimental results of the present invention show that after cigarette smoke modeling, the expression level of Bax mRNA in the lung tissue of mice increased significantly, and the level of Bcl2 mRNA decreased significantly, indicating that cigarette smoke caused an increase in lung tissue cell apoptosis to replace damaged lung tissue cells. When Physalis was given, the expression level of Bax mRNA in the lung tissue of mice was downregulated, and the level of Bcl2 mRNA was upregulated, indicating that the cell apoptosis process of the lung tissue was slowed down, and there was no need to compensate for the damage of lung tissue cells by promoting the formation of apoptotic cells.

[0068] In summary, Physalis has good antioxidant and anti-inflammatory effects, and can slow down the apoptosis of lung tissue cells and improve lung tissue damage caused by cigarette smoke.

[0069] The present invention discloses a Physalis extract and a preparation method thereof and use thereof in the preparation of a drug for treating lung injury caused by cigarette smoke. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve the purpose. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all deemed to be included in the present invention. The products of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the products described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

Claims

1. A method for preparing a Physalis alkekengi extract, characterized in that: The following steps are involved: Fresh physalis berries are added into pure water, and the berries are continuously concentrated by decoction to obtain a paste-like pulp. The initially concentrated paste-like pulp is placed into a freeze dryer for freeze-drying, and the dried product is collected. The solid physalis berries are then ground into fine powder using a mortar.

2. The method for preparing the Physalis alkekengi extract according to claim 1, characterized in that: The usage ratio of the fresh physalis berry to the pure water is in the range of 0.5-0.6 g / ml.

3. The method for preparing the Physalis alkekengi extract according to claim 1, characterized in that: In the decoction method, the decoction times range from 1 to 2 times, the total decoction time ranges from 3 to 4 hours, and the decoction temperature ranges from 90 to 100°C.

4. The method for preparing the Physalis alkekengi extract according to claim 1, characterized in that: The freeze-drying conditions are: temperature: -50 to -40°C, pressure: 1 to 10 Pa.

5. The method for preparing the Physalis alkekengi extract according to claim 1, characterized in that: The mesh size of the fine powder ranges from 60 to 100 meshes.

6. The Physalis alkekengi extract obtained by the method for preparing the Physalis alkekengi extract according to any one of claims 1 to 5.

7. Use of the Physalis alkekengi extract according to claim 6 in the preparation of a drug for treating lung damage caused by cigarette smoke.

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