Immune organ repairing pharmaceutical composition containing Wanshu capsule as active component

Abstract

The invention discloses a pharmaceutical composition containing Wanshu capsules as an active ingredient and used for repairing immune organs. The Wanchu capsule provided by the invention can obviously increase the proportion of peripheral blood lymphocytes.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine, and in particular to a pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient. Background Technology

[0002] Wanying Capsules are a commercially available traditional Chinese medicine, traditionally used to treat high fever, irritability, mouth ulcers, and swollen and sore gums and throat in children.

[0003] To date, there have been no reports of Wanying Capsules as an active ingredient in repairing immune organs.

[0004] The above background information is provided to facilitate understanding of the present invention and is not intended to be publicly known technology disclosed to the general public prior to the application of this invention. Summary of the Invention

[0005] To address the aforementioned deficiencies, the present invention provides a pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient, which aims to improve at least one of the problems mentioned in the background art.

[0006] The technical solution is: a pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient.

[0007] Furthermore, when Wanying Capsules are used as the active ingredient in a pharmaceutical composition for repairing immune organs, they are used in combination with cyclophosphamide; the way to repair immune organs is by regulating the proportion of lymphocytes to restore cellular immune function; the dosage of Wanying Capsules is 0.1142 g / kg•d~0.4569 g / kg•d.

[0008] Furthermore, the immune organ is the spleen.

[0009] Furthermore, the spleen is a mouse spleen.

[0010] Furthermore, the immune organ is the thymus.

[0011] Furthermore, the substance in question is the thymus of a mouse.

[0012] This invention also provides an application of the Wanying Capsule.

[0013] The technical solution is: the preparation of a pharmaceutical composition for repairing immune organs using a universal capsule.

[0014] Furthermore, the dosage of Wanying Capsules is 0.1142 g / kg•d to 0.4569 g / kg•d.

[0015] Furthermore, the immune organ is either the spleen or the thymus.

[0016] Furthermore, the spleen is a mouse spleen, and the thymus is a mouse thymus.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention found that pretreatment with Wanying Capsules can significantly increase the spleen index in mice, indicating that Wanying Capsules can improve spleen damage and thus enhance non-specific immunity. Wanying Capsules can also significantly increase the proportion of peripheral blood lymphocytes, suggesting that Wanying Capsules can alleviate the damage to cellular immune function in mice to a certain extent. Attached Figure Description

[0018] Figure 1 The changes in body weight of mice in each group (n=10) according to this invention; Figure 2 The images show the pathological sections of the spleen and thymus of mice in each group of mice in this invention (n=3). Detailed Implementation

[0019] As used in this article: "Prepared from" is synonymous with "comprising". The terms "comprising", "including", "having", "containing", or any other variations thereof as used herein are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0020] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range “1–5” is disclosed, the described range should be interpreted as including ranges “1–4”, “1–3”, “1–2”, “1–2 and 4–5”, “1–3 and 5”, etc. When numerical ranges are described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range.

[0021] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be construed as limiting the scope of this application. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0023] In these embodiments, unless otherwise specified, the portions and percentages are all by weight.

[0024] "Parts by mass" refers to the basic unit of measurement that expresses the mass ratio of multiple components. One part can represent any unit mass, such as 1g or 2.689g. If we say that component A has "a" parts by mass and component B has "b" parts by mass, it means the ratio of the mass of component A to the mass of component B is a:b. Alternatively, it can mean that the mass of component A is aK and the mass of component B is bK (K is any number representing a multiplier). It is important to understand that, unlike the number of parts by mass, the sum of the mass parts of all components is not limited to 100 parts.

[0025] "And / or" is used to indicate that one or both of the described situations may occur, for example, A and / or B includes (A and B) and (A or B).

[0026] In this invention, the experimental animals are: Sixty SPF-grade BALB / c mice, half male and half female, 8 weeks old, weighing (20±2) g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Production License No.: SCXK2025-0006). Husbandry conditions: 12-hour light / dark cycle, ambient temperature 22–25℃, relative humidity 30%–50%. All animals were acclimatized for 7 days before the start of the experiment. This experimental animal protocol has been reviewed and approved by the Scientific Research Ethics Committee of Shanxi University, and the experimental procedures were strictly carried out in accordance with the ethical review requirements for laboratory animals of Shanxi University (Approval No.: SXULL2025035).

[0027] In this invention, the experimental drug is: Wanying Capsules: Batch number 231103, specification 0.3 g × 6 capsules, adult daily clinical dosage is crude drug dose: 0.025 g / kg•d, based on an adult weight of 60 kg, provided by Chengdu Yangtian Wanying Pharmaceutical Co., Ltd.

[0028] Positive control Zhenqi Fuzheng Granules: batch number K20241017, specification 5 g / bag, adult daily clinical dosage is: 0.167 g / kg•d, based on an adult weight of 60 kg, Gansu Fuzheng Pharmaceutical Technology Co., Ltd.

[0029] In this invention, the experimental reagents include: cyclophosphamide, 4% paraformaldehyde solution, physiological saline, etc.

[0030] In this invention, the experimental equipment includes: surgical instruments, XDS-1B inverted microscope, etc.

[0031] In this invention, model preparation and group intervention are as follows: Mice were randomly divided into four groups based on body weight: a normal control group, a model control group, a group receiving Zhenqi Fuzheng Granules 1.5 g / kg•d (equivalent to a clinically equivalent dose), a high-dose group of Wanying Capsules (WYC) (0.4569 g / kg•d crude drug), a medium-dose group of Wanying Capsules (0.2284 g / kg•d crude drug), and a low-dose group of Wanying Capsules (0.1142 g / kg•d crude drug), with 10 mice in each group. Each group was administered the corresponding drug via gavage at a volume of 10 mL / kg for 14 consecutive days. The normal control group and the model control group received an equal volume of 0.5% sodium carboxymethyl cellulose. Except for the normal control group, the other groups received intraperitoneal injections of cyclophosphamide at 10 mL / kg per day for 9-12 days to establish an immunodeficient mouse model. The normal control group received an equal volume of 0.9% sodium chloride solution intraperitoneally.

[0032] In this invention, the indicator detection includes: 1. General Status Evaluation Macroscopic signs and behavioral changes in mice were observed and recorded before and after drug intervention.

[0033] 2. The influence of body weight and organ indices During the experiment, the body weight of mice in each group was recorded every two days, and changes were observed. One hour after the last administration, the weight of mice in each group was measured, blood was collected by enucleation, and the mice were euthanized by cervical dislocation. The spleen and thymus were dissected, and fat and fascia tissue were removed. The weight of the organs was measured, and the spleen index and thymus index were calculated using the following formulas: Spleen index = Spleen weight (mg) / Body weight (g); Thymus index = Thymus weight (mg) / Body weight (g).

[0034] 3. Measurement of peripheral blood parameters in mice After collecting blood from the mouse orbital cavity, an appropriate amount of whole blood was added to an EDTA-K2 anticoagulant tube, mixed well, and the number of white blood cells (WBC), lymphocytes (LYM), and neutrophils (NE) in the mouse peripheral blood was measured using an animal blood cell analyzer.

[0035] 4. HE histopathological analysis Six rats were randomly selected from each group 1 day and 7 days after drug administration and sacrificed. Oral mucosal specimens were obtained, rinsed with physiological saline, and immediately fixed in 4% paraformaldehyde solution. The tissues were embedded in paraffin, sectioned into 5 μm sections, stained with hematoxylin and eosin (HE), and then observed and analyzed under an optical microscope.

[0036] In this invention, experimental data are expressed as mean ± standard deviation, and statistical analysis was performed using SPSS 27.0 software. First, the data were tested for normality and homogeneity of variance. If the variances were homogeneous, one-way ANOVA was used for population comparison. If differences were found, the LSD method was used for pairwise comparisons between the means of multiple dose groups and a control group. If the variances were unequal, nonparametric tests were used. The significance level was set at α = 0.05. P < 0.05 was considered statistically significant.

[0037] The results of this invention are as follows: 1. General condition of the mice The control group mice were in good condition, active, and had shiny fur; the model group mice were lethargic, had dull fur, moved slowly, and preferred to huddle together and lie down. The general condition of the low, medium, and high dose Wanying capsules and the Zhenqi Fuzheng granules group was improved compared with that of the model group.

[0038] 2. Changes in body weight of mice in each group The changes in body weight of mice in each group during the experiment are as follows: Figure 1 As shown in the figure, compared with the control group, the body weight of mice in each model group decreased significantly after cyclophosphamide modeling, indicating a good modeling effect; while the body weight of the Zhenqi Fuzheng Granules group and the Wanying Capsule group gradually increased and tended to normal after drug intervention.

[0039] 3. Immune organ indices of mice in each group The results are shown in Table 1. Compared with the blank control group, the spleen index and thymus index of the model group decreased significantly, and the difference was statistically significant (P<0.01), indicating that the cyclophosphamide-induced immunosuppression model was successful. Compared with the model group, the spleen index and thymus index of the Zhenqi Fuzheng Granules group, Wanying Capsule group, and the three dosage groups increased significantly, and the difference was statistically significant (P<0.05, P<0.01), indicating that the intervention of Zhenqi Fuzheng Granules and Wanying Capsule had a certain ameliorative effect on cyclophosphamide-induced damage to the immune organs of mice. Compared with the Zhenqi Fuzheng Granules group, the high-dose Wanying Capsule group had a significant repair effect on the spleen and thymus of immunocompromised mice.

[0040] Table 1 Note: Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0041] 4. Changes in the number of peripheral blood biomarkers in each group of mice The results are shown in Table 2. Compared with the blank group, the number of WBCs and LYMs in the immunosuppressed model group mice was significantly reduced, while the number of NEs was significantly increased, and the differences were statistically significant (P < 0.01). Compared with the model group, the number of WBCs and LYMs was significantly increased and the number of NEs was significantly decreased after intervention with Wanying capsules, and the differences were statistically significant (P < 0.05, P < 0.01).

[0042] Table 2 Comparison of peripheral blood parameters in mice of different groups (n=10) Note: Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0043] 5. HE staining analysis Thymus microscopy revealed decreased cortical thickness, characterized by a reduction in cortical lymphocytes. Compared to group M, group WH showed better results, with significant improvement in the reduction of thymic lymphocytes. Splenomegaly revealed overall spleen atrophy, characterized by a significant reduction in lymphocytes in both the white and red pulp, and a decrease in spleen volume; in some milder cases, the periphery of the white pulp lymphatic sheath was obliterated. Results are shown in Table 3.

[0044] The spleen's tissue structure gradually returned to normal, the boundary between the red and white pulp became clearer, and there was a significant separation between the thymic cortex and medulla. Furthermore, the Wanying capsule group showed better results than the positive control group. Figure 2 In the diagram, Figure A represents the spleen, and Figure B represents the thymus. R represents the red pulp; W represents the white pulp; C represents the cortex; and M represents the medulla.

[0045] Table 3. Microscopic analysis results of histopathological features of mice in each group (n=3) The model mice of this invention showed decreased spleen index and a reduced number of peripheral blood leukocytes (mainly lymphocytes), suggesting that cyclophosphamide caused damage to both the non-specific and specific immune systems in mice. Literature review revealed that Zhenqi Fuzheng Granules have a significant effect on enhancing immune function; therefore, they were used as a positive control in this study.

[0046] The spleen is a major immune organ, and the spleen index can reflect the function of the body's non-specific immune system to a certain extent. Pretreatment with Wanying capsules significantly increased the spleen index in mice, indicating that Wanying capsules have the effect of improving spleen damage and thus enhancing non-specific immunity.

[0047] Leukocytes, differentiated from pluripotent hematopoietic stem cells, protect the body from pathogen invasion and tissue damage. Cyclophosphamide has direct anti-proliferative and pro-apoptotic effects on various cell types. In this study, cyclophosphamide reduced the number of peripheral blood leukocytes in mice, mainly manifested as a decrease in the number of peripheral blood lymphocytes. Wanying capsules significantly increased the proportion of peripheral blood lymphocytes, suggesting that Wanying capsules can alleviate impaired cellular immune function in mice to some extent.

[0048] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

[0049] Furthermore, those skilled in the art will understand that although some embodiments herein include certain features included in other embodiments but not others, combinations of features from different embodiments are intended to be within the scope of this application and form different embodiments. The information disclosed in this background section is intended only to enhance the understanding of the general background of this application and should not be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

Claims

1. A pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient.

2. The pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient according to claim 1, characterized in that, When Wanying Capsules are used as the active ingredient in a pharmaceutical composition for repairing immune organs, they are used in combination with cyclophosphamide. The way to repair immune organs is to regulate the proportion of lymphocytes and restore cellular immune function; the dosage of Wanying capsules is 0.1142 g / kg•d to 0.4569 g / kg•d.

3. The pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient according to claim 2, characterized in that, The immune organ in question is the spleen.

4. The pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient according to claim 3, characterized in that, The spleen in question is that of a mouse.

5. The pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient according to claim 2, characterized in that, The immune organ in question is the thymus.

6. The pharmaceutical composition for repairing immune organs containing Wanying Capsules as an active ingredient according to claim 5, characterized in that, The substance in question is the thymus of a mouse.

7. The preparation of a pharmaceutical composition for the use of a universal capsule in repairing immune organs.

8. In the preparation of the pharmaceutical composition for repairing immune organs using the Wanying Capsule according to claim 7, characterized in that, The dosage of Wanying Capsules is 0.1142 g / kg•d to 0.4569 g / kg•d.

9. In the preparation of the pharmaceutical composition for repairing immune organs using the Wanying Capsule according to claim 7, it is characterized in that, The immune organ is either the spleen or the thymus.

10. In the preparation of the pharmaceutical composition for repairing immune organs using the Wanying Capsule according to claim 9, it is characterized in that, The spleen is the spleen of a mouse, and the thymus is the thymus of a mouse.

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