Application of Dayuan drink extract in preparation of medicine for treating human coronavirus 229E infected pneumonia

Abstract

The invention belongs to the technical field of medicines, and discloses application of a Dayuan drink extract in related medicines for treating human coronavirus 229E infected pneumonia. Specifically, the invention discloses application of a Dayuan drink extract in preparation of drugs for protecting respiratory dysfunction, lung injury, inflammatory response, virus infection and immune disorder caused by human coronavirus 229E infected pneumonia. Pharmacological experiments prove that the Dayuan drink extract can improve the respiratory function in human coronavirus 229E infected pneumonia, exert anti-inflammatory, antiviral and immunoregulation effects, and can improve pulmonary edema, thereby achieving the purpose of treatment.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a use of a Dayuanyin extract in preparing a medicament for preventing and / or treating pneumonia infected with human coronavirus 229E. Background Art

[0002] Human coronavirus 229E (HCoV-229E) is an alphacoronavirus and is one of the most important causes of respiratory health. It is widely spread among the population and is the main cause of the most common respiratory diseases in children in my country [1-3]. Although the symptoms of common coronavirus infections are mild and can resolve on their own, they can cause serious or even fatal diseases in children, the elderly, and immunocompromised patients [4], and can cause lower respiratory tract infections, leading to bronchiolitis and pneumonia [5].

[0003] Chinese herbal medicine has become an increasingly popular treatment method due to its good effectiveness and low side effects. Dayuanyin comes from "Treatise on Warm Epidemics" written by Wu Youxing in the Ming Dynasty. It is a classic prescription for treating lung diseases. It is composed of seven herbs: betel nut, tsaoko, peony, magnolia bark, scutellaria, rhizoma anemarrhenae and licorice. It is an important prescription for treating the initial stage of warm epidemics when evil is lurking in the membrane, and it is also an important prescription for treating symptoms of dampness lurking in the membrane. It has the function of opening up the membrane, eliminating filth and clearing turbidity. Since its introduction, this prescription has received widespread attention in the medical community. Zhang Ru et al. reported that Dayuanyin may reduce lung damage caused by respiratory syncytial virus infection by improving lung tissue damage in mice, reducing inflammation, and regulating gastrointestinal hormone levels and lymphocyte percentages. [8] Wang Lifeng et al. reported that Xinjiadayuan Drink has a significant anti-hepatitis B virus effect in vitro[9].

[0004] Sun Weiguang et al. explored the inhibitory effect of Juhong Tanke Granules on human coronavirus HCoV-229E

[10] . Peng Qing et al. reported that Majingxuanfei Granules can play a therapeutic role in mice with human coronavirus 229E pneumonia by inhibiting coronavirus replication, inhibiting the release of inflammatory factors, and regulating immune balance

[11] . Yu Xiangcui et al. revealed the pharmacodynamic mechanism and potential active ingredients of Xiaochaihu Granules against HCoV-229E. Its antiviral mechanism may be related to inhibiting the overexpression of inflammatory factors, inhibiting viral replication, invasion, and transcription.

[12] Geng Zihan et al. reported that Gegentang granules can improve the general signs of mice with HCoV-229E cold-dampness epidemic toxin attacking the lungs, regulate gastrointestinal hormone levels, reduce lung index and lung tissue pathology scores, and may exert an immunomodulatory effect by inhibiting inflammatory cytokines in lung tissue and restoring the percentage of peripheral blood lymphocytes, thereby having a therapeutic effect on model mice.

[13] In vitro experiments by Liu Zhenhai et al. showed that Yinqiao Jiedu Soft Capsules can inhibit the replication of human coronavirus HCoV-229E and its mediated inflammatory response, and have the development value of anti-coronavirus drugs.

[14] .

[0005] In Chinese patent CN115919921A, Guangzhou Baiyunshan Huacheng Pharmaceutical Co., Ltd. discloses the use of a monkey earring extract in the preparation of a drug for preventing the proliferation of coronavirus, the coronavirus being HCoV-229E

[15] In Chinese patent CN113398235A, Guangzhou Baiyunshan Guanghua Pharmaceutical Co., Ltd. discloses the use of a traditional Chinese medicine composition in the preparation of a drug for preventing and treating human coronavirus HCoV-229E. The active ingredients of the traditional Chinese medicine composition are prepared from bupleurum, scutellaria, pinellia, codonopsis, ginger, licorice, and jujube.

[16] In Chinese patent CN111110701B, Lei Yunshang Pharmaceutical Group Co., Ltd. disclosed a new use of Liushen Pills in the preparation of drugs for preventing and treating viral diseases.

[17] In Chinese patent CN111150755B, Lei Yunshang Pharmaceutical Group Co., Ltd. disclosed a new use of a Chinese herbal medicine composition in the preparation of drugs for preventing and treating viral diseases, which is composed of Patchouli, Chrysanthemum indicum, Elsholtzia cava and Artemisia annua.

[18] In Chinese patent CN116139195A, Jiangxi University of Traditional Chinese Medicine and Jiangxi Guxiangjinyun Health Industry Co., Ltd. disclosed an anti-coronavirus compound essential oil composition, its preparation method and use. The anti-coronavirus compound essential oil composition comprises the following raw materials by weight: volatile oil of Chimonanthus chinensis leaves (10-20 parts), volatile oil of cloves (5-10 parts), volatile oil of mugwort (5-10 parts), volatile oil of Elsholtzia ciliata (3-5 parts), volatile oil of Perilla frutescens (3-5 parts), volatile oil of peppermint (2-5 parts), eucalyptus essential oil (3-5 parts), geranium essential oil (2-5 parts), lavender essential oil (2-5 parts), ravensa essential oil (2-5 parts), laurel essential oil (2-5 parts), and tea tree essential oil (3-5 parts).

[19] In Chinese patent CN114712453A, Shijiazhuang Yiling Pharmaceutical Co., Ltd. discloses a Chinese herbal medicine composition for use in antiviral, organ protection, and immunity enhancement drugs. The composition mainly consists of ephedra, gypsum, forsythia, scutellaria, mulberry bark, bitter almond, and peucedanum.

[20] In Chinese patent CN115089634A, Shanxi Badun Environmental Protection Technology Research Institute disclosed a bezoar / essential oil composition for preventing and treating COVID-19, as well as its preparation method and use. The composition contains bezoar and four essential oil components, including sweet orange oil, forsythia oil, carvacrol, passion fruit oil, and

[21] In Chinese patent CN113209185B, Jiangxi University of Traditional Chinese Medicine discloses a broad-spectrum antiviral Chinese medicine volatile oil preparation and its use, including tea tree volatile oil, mountain sweet volatile oil, thyme volatile oil, bay leaf volatile oil, eucalyptus volatile oil, lemon volatile oil, houttuynia cordata volatile oil, myrtle volatile oil, ribwort leaf volatile oil, Atlantic cedar volatile oil, wormwood volatile oil.

[22] .

[0006] The aforementioned patents cover traditional Chinese medicine formulas or compositions currently used to treat pneumonia caused by human coronavirus 229E, which differ fundamentally from the Dayuanyin compound described in the present invention. Compared to other traditional Chinese medicine compounds for treating lung diseases, Dayuanyin's treatment of viral pneumonia is characterized by: regulating viral-host interactions, inhibiting viral infection and replication; enhancing the host's ability to resist viral disease and clear the virus; and enhancing the host's ability to resist disease progression, halting disease progression, and promoting host repair. Therefore, through examination of relevant indicators, it was found that Dayuanyin extract has therapeutic effects on pneumonia caused by human coronavirus 229E. In summary, there are no reports to date on the therapeutic effects of the Dayuanyin compound on pneumonia caused by human coronavirus 229E. In light of this, the present invention is proposed. Summary of the Invention

[0007] The object of the present invention is to provide a new use of Dayuanyin extract in the preparation of a medicament for preventing and / or treating pneumonia infected with human coronavirus 229E.

[0008] One of the purposes of the present invention is to provide the use of Dayuanyin extract in related medicines and other products for the treatment of pneumonia infected with human coronavirus 229E.

[0009] The second purpose of the present invention is to provide the use of Dayuanyin extract in treating symptoms such as inflammatory response, immune disorder, viral infection, and abnormal respiratory function caused by pneumonia infected with human coronavirus 229E.

[0010] The third purpose of the present invention: The Dayuanyin extract of the present invention can effectively restore the increase in respiratory rate (F) and minute ventilation (MV) and the decrease in peak expiratory flow (PEF) and enhanced pause (PenH) caused by human coronavirus 229E pneumonia, thereby improving respiratory function.

[0011] The fourth purpose of the present invention: The Dayuanyin extract of the present invention achieves anti-inflammatory effects by reducing the number of white blood cells (WBC) and neutrophils (NEUT) in the blood, and reducing the levels of interleukin-1β (IL-1β), interleukin-10 (IL-10), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the serum.

[0012] The fifth purpose of the present invention: The Dayuanyin extract of the present invention can reduce the lung index, restore the alveolar state of damaged lung tissue, and achieve the effect of repairing lung tissue.

[0013] The sixth purpose of the present invention: The Dayuanyin extract of the present invention can reduce the viral load of lung tissue and achieve an antiviral effect.

[0014] The seventh purpose of the present invention: The Dayuanyin extract of the present invention can reduce the number of lymphocytes (LYM) in the blood, adjust the spleen index and thymus index, increase the CD4 / CD8 value and the number of antigen presenting cells (APC), and achieve immune regulation.

[0015] In order to solve the technical problems of the present invention, the present invention provides the following technical solutions:

[0016] The technical solution of the present invention provides a Dayuanyin extract, wherein every 100 parts by weight of the formula is prepared from the following raw materials by weight: 10-40 parts by weight of betel nut, 5-20 parts by weight of magnolia bark, 1-10 parts by weight of tsaoko, 10-20 parts by weight of white peony root, 5-20 parts by weight of anemarrhena, 8-20 parts by weight of scutellaria, and 1-15 parts by weight of liquorice.

[0017] It is further preferred that every 100 parts by weight of the Dayuanyin extract is made from the following raw materials in parts by weight: 28 parts by weight of betel nut, 14 parts by weight of magnolia bark, 8 parts by weight of tsaoko, 14 parts by weight of white peony root, 14 parts by weight of anemarrhena, 14 parts by weight of scutellaria, and 8 parts by weight of liquorice.

[0018] The technical solution of the present invention uses a heating reflux extraction method to extract Dayuan Drink powder: take the above-mentioned medicinal materials in parts by weight, add 5, 4, and 3 times the amount of water in sequence, heat reflux extraction 3 times, each time for 1.5 hours, filter and combine the filtrate, concentrate under reduced pressure, and dry at 70°C for 48 hours to obtain Dayuan Drink powder.

[0019] In the above method, preferably, the extraction reflux times are 3 times, and the extraction reflux time is 1.5 hours;

[0020] A mouse model of pneumonia caused by human coronavirus 229E was established using male Balb / c mice infected intranasally with HCoV-229E virus solution. Mice were infected twice, on the first and third days. Drug administration began on the first day of infection. Daily weighing and blood sampling were performed during drug administration. Respiratory function was measured on the second and fourth days of the experiment. After four days of drug treatment, serum was collected to measure inflammatory cytokine levels. The lungs, spleen, and thymus were weighed and weighed to calculate organ indices. Partial lung tissue was obtained for viral nucleic acid load determination. Flow cytometry was used to determine the percentage of splenic lymphocytes. The therapeutic effect of Dayuanyin extract on pneumonia caused by human coronavirus 229E was evaluated.

[0021] The present invention also relates to pharmaceutical compositions of the compounds of the present invention and conventional pharmaceutical excipients or adjuvants.

[0022] The pharmaceutical composition of the compounds of this invention can be prepared according to methods well known in the art. When used for this purpose, if necessary, the compounds of this invention can be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to prepare suitable administration forms or dosage forms that can be used as human or veterinary medicines.

[0023] The compound of the present invention or the pharmaceutical composition containing the same can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum or rectum.

[0024] The compound of the present invention or the pharmaceutical composition containing the same can be administered by injection, including intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and acupuncture injection.

[0025] The dosage form can be a liquid or solid dosage form. For example, liquid dosage forms can be true solutions, colloids, microparticles, emulsions, or suspensions. Other dosage forms include tablets, capsules, pellets, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, and freeze-dried powder injections.

[0026] The compound of the present invention can be prepared into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle delivery systems.

[0027] Beneficial Technical Effects: This compound exhibits outstanding pharmacological activity. Through the synergistic effects of traditional Chinese medicine, it effectively restores the increased respiratory rate and minute ventilation, as well as the decreased peak expiratory flow and enhanced expiratory interval, caused by pneumonia caused by human coronavirus 229E. It also reduces the number of white blood cells, lymphocytes, and neutrophils in the blood, and the levels of IL-1β, IL-10, IL-6, and TNF-α in serum. It also lowers the lung index, increases the spleen index and thymus index, reduces the viral load in lung tissue, and increases the CD4 / CD8 ratio and APC count. This results in synergistic effects of improving respiratory function, anti-inflammatory and antiviral effects, lung tissue repair, and immune regulation. The extract is suitable for the preparation of a drug for the prevention and / or treatment of pneumonia caused by human coronavirus 229E. The extract is highly safe, has a reliable ingredient source, and is low in production cost, offering promising market development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Effects of Dayuanyin extract on respiratory function in mice with pneumonia infected with human coronavirus 229E;

[0029] Figure 2 Effects of Dayuanyin extract on routine blood parameters in mice with pneumonia infected with human coronavirus 229E;

[0030] Figure 3 Effects of Dayuanyin extract on organ indexes in mice with pneumonia infected with human coronavirus 229E;

[0031] Figure 4 Effects of Dayuanyin extract on serum inflammatory factors in mice with pneumonia infected with human coronavirus 229E;

[0032] Figure 5 Effects of Dayuanyin extract on the viral load in lung tissue of mice with pneumonia infected with human coronavirus 229E;

[0033] Figure 6 Effects of Dayuanyin extract on spleen lymphocytes in mice with pneumonia infected with human coronavirus 229E; Figure 7 Effects of Dayuanyin extract on damaged lung tissue in mice with pneumonia infected with human coronavirus 229E DETAILED DESCRIPTION

[0034] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention will be described in further detail below with reference to the accompanying drawings.

[0035] Table 1 Abbreviations

[0036]

[0037]

[0038] Example 1

[0039] Preparation of Dayuanyin Extract by Heating Reflux Extraction

[0040] Dayuanyin extract was prepared by reflux extraction method, according to 28 parts by weight of betel nut, 14 parts by weight of magnolia bark, 8 parts by weight of tsaoko, 14 parts by weight of white peony root, 14 parts by weight of anemarrhena, 14 parts by weight of scutellaria, and 8 parts by weight of liquorice:

[0041] (1) Weigh betel nut, Magnolia officinalis, Anemarrhena asphodeloides, White Peony Root, Scutellaria baicalensis, Licorice, and Amomum villosum according to the above weight ratio, mix well, and place in a 1000 mL round-bottom flask;

[0042] (2) Add 5, 4, and 3 times the amount of water in sequence, and heat under reflux for extraction 3 times, each time for 1.5 hours;

[0043] (3) Filter and combine the filtrates, concentrate under reduced pressure, and dry at 70° C. for 48 hours to obtain Dayuan drink powder.

[0044] Example 2

[0045] Effects of Dayuanyin extract on respiratory function in mice with pneumonia infected with human coronavirus 229E

[0046] 1. Experimental Methods

[0047] Embodiment Animal selection male SPF level Balb / c mice 40, body weight 12-14g, are randomly divided into 5 groups, 8 in every group, and above-mentioned 5 groups are respectively set to normal control group, model group, Dayuanyin extract low, medium and high three dosage groups (0.3g / kg, 1g / kg, 3g / kg).Give corresponding dosage medicine with gavage mode, namely start administration after first day modeling, administration four days.On the second day of modeling and the 4th day, 6 mice of each group adopt WBP whole body plethysmography system to measure respiratory function, after instrument calibration is zeroed, arranging flow parameter is 0.3L / min, mice is put into adaptation 15min in advance, and then record 15min and be measured data.

[0048] 2. Experimental Results

[0049] Compared with the normal group mice, the F and MV of the model group mice increased significantly on the second and fourth days after modeling, while the peak expiratory flow PEF and PENH decreased on the second day and significantly decreased on the fourth day ( ## P<0.01, ###P<0.001). However, treatment with 3g / kg Dayuanyin effectively reduced F and MV and significantly increased PEF, and the effect was more significant on the fourth day. PenH was also effectively increased on the fourth day. Treatment with 1g / kg Dayuanyin significantly reduced F and increased PEF on the second day, and significantly reduced F and MV and significantly increased PEF on the fourth day, but had no significant effect on PenH. Treatment with 0.3g / kg Dayuanyin significantly increased PEF on the second day, and effectively reduced F and MV and significantly increased PEF and PenH on the fourth day. * P<0.05, ** P<0.01, *** P<0.001). The positive drug Lianhua Qingwen effectively reduced F and MV and significantly increased PEF, and the effect was more obvious on the fourth day ( Figure 1 , Table 2).

[0050] Table 2 Effects of Dayuanyin extract on respiratory function of mice with pneumonia infected with human coronavirus 229E (Mean±SD)

[0051]

[0052] Mean ± SD, n = 6; ## P<0.01, ### P<0.001vs normal group, * P<0.05, ** P<0.01 *** P<0.001 vs model group

[0053] Example 3

[0054] Effects of Dayuanyin extract on routine blood parameters in mice with pneumonia infected with human coronavirus 229E

[0055] 1. Experimental Methods

[0056] Examples Animals 40 male SPF Balb / c mice weighing 12-14 g were randomly divided into 5 groups of 8 mice each. The 5 groups were set as a normal control group, a model group, and three dose groups (0.3 g / kg, 1 g / kg, and 3 g / kg) of Dayuanyin extract at low, medium, and high doses. The corresponding doses of drugs were administered by gavage. After modeling on the first day, drug administration began. For four days, 20 μL of blood was collected four hours after administration every day and tested using a hematology five-type cardinality analyzer for five consecutive days.

[0057] 2. Experimental Results

[0058] Compared with the normal group, the number of white blood cells, lymphocytes and neutrophils in the model group increased significantly, and it was more significant from the second to the fourth day ( # P<0.05, ##P<0.01, ### P<0.001). Treatment with 3g / kg Dayuanyin can significantly reduce the number of white blood cells, lymphocytes and neutrophils, and the effect of reducing the number of neutrophils is more obvious on the third and fourth days; treatment with 1g / kg Dayuanyin can significantly reduce the number of white blood cells, lymphocytes and neutrophils, and the effect of reducing lymphocytes is reduced at the second infection on the third day, and the effect of reducing the number of neutrophils is more obvious on the fourth day; treatment with 0.3g / kg Dayuanyin can significantly reduce the number of white blood cells, lymphocytes and neutrophils, and the effect of reducing the number of white blood cells and lymphocytes is more obvious on the fourth day, and the effect of reducing the number of neutrophils is significant on the second and fourth days, and more obvious on the fourth day ( * P<0.05, ** P<0.01, *** P < 0.001)( Figure 2 , Table 3).

[0059] Table 3 Effects of Dayuanyin extract on routine blood indicators in mice with pneumonia infected with human coronavirus 229E (Mean±SD)

[0060]

[0061] Mean±SD,n=6, # P<0.05, ## P<0.01, ### P<0.001vs normal group, * P<0.05, ** P<0.01,

[0062] *** P<0.001 vs model group

[0063] Example 4

[0064] Effects of Dayuanyin extract on organ indexes in mice with pneumonia infected with human coronavirus 229E

[0065] 1. Experimental Methods

[0066] Examples Animals 40 male SPF Balb / c mice, weighing 12-14 g, were randomly divided into 5 groups of 8 mice each. The 5 groups were set as a normal control group, a model group, and three dose groups (0.3 g / kg, 1 g / kg, and 3 g / kg) of Dayuanyin extract at low, medium, and high doses, respectively. The corresponding doses of drugs were administered by gavage. Dosing began on the first day after modeling. The mice were weighed on the fifth day after administration for four days. After blood was drawn, the lungs, spleen, and thymus were washed with physiological saline to flush excess blood. The excess water was then absorbed with filter paper. The weighed weights were calculated as organ weights. Organ index (%) = organ weight (g) / body weight (g) × 100%.

[0067] 2. Experimental Results

[0068] Compared with the normal group, the lung index of the model group mice increased significantly, and the thymus and spleen indexes decreased significantly ( # P<0.05, ## P<0.01, ### P<0.001). Treatment with 3g / kg Dayuanyin effectively reduced the lung index and significantly increased the thymus index and spleen index; treatment with 1g / kg Dayuanyin significantly reduced the lung index, but had no significant effect on the thymus and spleen indexes; treatment with 0.3g / kg Dayuanyin effectively reduced the lung index and significantly increased the spleen index, but had no significant effect on the thymus index ( * P<0.05, ** P<0.01, *** P < 0.001)( Figure 3 , Table 4).

[0069] Table 4 Effects of Dayuanyin extract on organ indexes of mice with pneumonia infected with human coronavirus 229E (Mean±SD)

[0070]

[0071] Mean±SD,n=8, ## P<0.01, ### P<0.001vs normal group, * P<0.05, ** P<0.01 *** P<0.001 vs model group

[0072] Example 5

[0073] Effects of Dayuanyin extract on serum inflammatory factors in mice with pneumonia infected with human coronavirus 229E

[0074] 1. Experimental Methods

[0075] Embodiment Animal selection Male SPF grade Balb / c mice 40, body weight 12-14g, were randomly divided into 5 groups, 8 in each group, and the above 5 groups were set as normal control group, model group, and Dayuanyin extract low, medium and high dose groups (0.3g / kg, 1g / kg, 3g / kg). The corresponding dose of medicine was given by gavage, and the drug was started after modeling on the first day. Blood was collected for four days after administration. The blood was loaded into a 1.5mL EP tube, placed at room temperature for 2h, put into a 4°C pre-cooled low-temperature centrifuge, and centrifuged at 3000rpm for 15min. The serum was divided into EP tubes and stored at -80°C to be tested. The content of inflammatory factors in serum was detected using ultrasensitive multifactor electrochemiluminescence technology.

[0076] 2. Experimental Results

[0077] Compared with the normal group, the serum levels of TNF-α, IL-10 and IL-1β in the model group mice were significantly increased ( # P<0.05, ## P<0.01, ### P<0.001). The treatment of Dayuanyin significantly reduced the levels of TNF-α, IL-10 and IL-1β in serum ( * P<0.05, ** P<0.01, *** P < 0.001)( Figure 4 , Table 5).

[0078] Table 5 Effects of Dayuanyin extract on serum inflammatory factors in mice with pneumonia infected with human coronavirus 229E (Mean±SD)

[0079]

[0080] Mean±SD,n=8, ## P<0.01, ### P<0.001vs normal group, *** P<0.001 vs model group

[0081] Example 6

[0082] Effect of Dayuanyin extract on viral load in lung tissue of mice with pneumonia infected with human coronavirus 229E

[0083] 1. Experimental Methods

[0084] Example Animals 40 male SPF Balb / c mice weighing 12-14g were randomly divided into 5 groups of 8 mice each. The 5 groups were set as normal control group, model group, and Dayuanyin extract low, medium and high dose groups (0.3g / kg, 1g / kg, 3g / kg). The corresponding dose of the drug was given by gavage. The drug was started after modeling on the first day. After four days of administration, the lung tissue was subpackaged and stored at -80°C. 5mg of lung tissue was ground and RNA was extracted. The lung tissue nucleic acid load was determined according to the human coronavirus 229E fluorescent quantitative RT-PCR detection kit.

[0085] 2. Experimental Results

[0086] Compared with the normal group mice, the viral load in the lung tissue of the model group mice was significantly increased ( ### P<0.001). The viral load in lung tissue of each group treated with Dayuanyin was significantly increased ( * P<0.05, ** P < 0.01)( Figure 5 , Table 6).

[0087] Table 6 Effect of Dayuanyin extract on the viral load of lung tissue in mice infected with human coronavirus 229E pneumonia (Mean±SD)

[0088]

[0089] Mean±SD,n=3, ### P<0.001vs normal group, * P<0.05, ** P<0.01 vs model group

[0090] Example 7

[0091] Effects of Dayuanyin extract on spleen lymphocytes in mice with pneumonia infected with human coronavirus 229E

[0092] 1. Experimental Methods

[0093] Embodiment Animal selection Male SPF grade Balb / c mice 40, body weight 12-14g, were randomly divided into 5 groups, 8 mice in each group, and the above 5 groups were respectively set as normal control group, model group, and three dose groups (0.3g / kg, 1g / kg, 3g / kg) of Dayuanyin extract at low, medium and high doses. The corresponding dose of medicine was given by gavage. After modeling on the first day, administration was started. After four days of administration, spleen was taken and 1640 culture medium was added and ground and centrifuged (3000rpm, 10min) to obtain splenocytes. After resuspension, red blood cell lysis solution was added and fully lysed and then centrifuged (3000rpm, 10min). The precipitation was resuspended and centrifuged (3000rpm, 10min) with normal saline. This was repeated twice. After staining with CD3, CD4, CD8 and CD19 antibodies, spleen lymphocytes were detected by flow cytometry.

[0094] 2. Experimental Results

[0095] Compared with the normal group, the CD4 / CD8 value and APC number of the model group mice were significantly decreased ( # P<0.05, ### P<0.001). Treatment with 0.3 and 3 g / kg Dayuanyin significantly increased the CD4 / CD8 value and APC number, and treatment with 1 g / kg Dayuanyin significantly increased the CD4 / CD8 value ( * P<0.05, ** P<0.01, *** P < 0.001)( Figure 6 , Table 7).

[0096] Table 7 Effect of Dayuanyin extract on spleen lymphocytes of mice infected with human coronavirus 229E pneumonia (Mean±SD)

[0097]

[0098] Mean±SD,n=3, # P<0.05 ### P<0.001vs normal group, * P<0.05, ** P<0.01 vs. Model Group Example 8

[0099] Effects of Dayuanyin extract on damaged lung tissue in mice with pneumonia infected with human coronavirus 229E

[0100] 1. Experimental Methods

[0101] Examples Animals Forty male SPF Balb / c mice weighing 12-14 g were randomly divided into five groups of eight mice each. The five groups were set as a normal control group, a model group, and three dose groups (0.3 g / kg, 1 g / kg, and 3 g / kg) of Dayuanyin extract at low, medium, and high doses, respectively. The corresponding doses of drugs were administered by gavage. Dosing began on the first day after modeling. Dosing continued for four days. On the fifth day, the left lungs of each group of mice were dissected, fixed with fixative, dehydrated, embedded in paraffin, and sectioned for H&E staining.

[0102] 2. Experimental Results

[0103] By comparing the lung tissue sections after staining, compared with the control group, the model group had disordered alveolar structure, ruptured alveolar septa, dilated alveoli, and a significantly reduced number of alveoli, while the above pathological changes in the lung tissue of mice in the Dayuanyin extract intervention group were improved ( Figure 7 ).

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[0119]

[15] Chinese patent CN115919921A

[0120]

[16] Chinese patent CN113398235A

[0121]

[17] Chinese patent CN111110701B

[0122]

[18] Chinese patent CN111150755B

[0123]

[19] Chinese patent CN116139195A

[0124]

[20] Chinese patent CN114712453A

[0125]

[21] Chinese patent CN115089634A

[0126]

[22] Chinese patent CN113209185B

Claims

1. Use of a Dayuanyin extract in the preparation of a medicament for preventing and / or treating pneumonia caused by human coronavirus 229E infection.

2. According to the use described in claim 1, the use of Dayuanyin extract in treating inflammatory response, immune disorder, viral infection, and respiratory dysfunction symptoms caused by human coronavirus 229E pneumonia is provided.

3. The use according to claim 1, characterized in that Dayuanyin extract can achieve the therapeutic effect of pneumonia caused by human coronavirus 229E infection by improving respiratory function.

4. The use according to claim 3, characterized in that Dayuanyin extract effectively restores the increase in respiratory rate and minute ventilation and the decrease in peak expiratory flow and enhanced expiratory interval caused by human coronavirus 229E pneumonia, and improves respiratory function.

5. The use according to claim 1, characterized in that Dayuanyin extract can achieve the therapeutic effect of pneumonia caused by human coronavirus 229E infection through anti-inflammatory effect.

6. The use according to claim 5, characterized in that Dayuanyin extract achieves anti-inflammatory effects by reducing the number of white blood cells and neutrophils in the blood, and lowering the levels of interleukin-1β, interleukin-10, interleukin-6, and tumor necrosis factor-α in the serum.

7. The use according to claim 1, characterized in that Dayuanyin extract can treat pneumonia caused by human coronavirus 229E infection by repairing damaged lung tissue and reducing pulmonary edema.

8. The use according to claim 7, characterized in that Dayuanyin extract can reduce the lung index, restore the alveolar state of damaged lung tissue, and achieve lung tissue repair.

9. The use according to claim 1, characterized in that Dayuanyin extract achieves therapeutic effect on pneumonia caused by human coronavirus 229E through antiviral effect.

10. The use according to claim 9, characterized in that Dayuanyin extract achieves its antiviral effect by reducing the viral load in lung tissue.

11. The use according to claim 1, characterized in that Dayuanyin extract achieves the therapeutic effect of pneumonia infected with human coronavirus 229E through immunomodulatory effect.

12. The use according to claim 11, characterized in that Dayuanyin extract achieves immune regulation by reducing the number of lymphocytes in the blood, adjusting the spleen index and thymus index, and increasing the CD4 / CD8 value and the number of antigen-presenting cells.

13. Use of a pharmaceutical composition for preventing and / or treating pneumonia caused by human coronavirus 229E infection, characterized in that: The pharmaceutical composition comprises the Dayuanyin extract according to claim 1 and a pharmaceutically acceptable carrier or excipient.

14. Use according to claim 13, characterized in that The dosage form of the composition preparation is selected from: oral preparation, injection dosage form or skin mucosal administration dosage form, oral preparation includes tablets, sustained-release preparations, capsules, controlled-release preparations, enteric solvents, pills or liquid preparations, and the injection dosage form includes intramuscular injection, intravenous injection or intravenous drip.

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