CN115887593B - Application of qinggongshou peach pill in preparing medicine for regulating autoimmune property - Google Patents
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Abstract
The invention provides an application of qinggong shou peach pills in preparing an autoimmune regulation medicament, which explores the prevention and treatment effects of the qinggong shou peach pills on the autoimmune diseases through a collagen-adjuvant induced rheumatoid arthritis model of the autoimmune diseases of rats, lays a foundation for the research on the prevention and treatment effects of the qinggong shou peach pills on the autoimmune diseases, and provides clues and basis for the application of the qinggong shou peach pills in the aspect.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an application of a palace-clearing birthday peach pill in preparation of an autoimmune regulation medicine.
Background
The palace-clearing life peach pill belongs to a secret prescription of Qianlong annual palace, the original prescription is named as flat peach pill, and the prescription consists of donkey kidney, deer kidney, dog kidney, medlar, ginseng, asparagus root, dwarf lilyturf tuber, rehmannia root, chinese angelica, sharpleaf galangal fruit (salted), silkworm excrement, wild jujube seed (fried) and split wood (fried coke), and belongs to a precious Chinese patent medicine in tonic medicines; the medicines are combined, so that the traditional Chinese medicine composition has the functions of tonifying kidney, producing sperm, soothing liver, strengthening spleen, promoting blood circulation, reducing phlegm, soothing nerves and improving intelligence, and mainly reduces or eliminates common aging symptoms of the elderly, such as dizziness, tiredness, hypomnesis, soreness of waist and knees, tinnitus, deafness, tearing, frequent nocturia, dribbling urine and the like; the pill is used after differentiation of symptoms of traditional Chinese medicine, and has certain improving effect on sexual hypofunction symptoms such as male impotence, premature ejaculation, hyposexuality and the like.
Rheumatoid arthritis belongs to an autoimmune disease mainly comprising systemic chronic diseases, and is mainly characterized by hyperplasia of synovial lining cells, infiltration of a large amount of inflammatory cells in the interstitium, angiogenesis of micro blood vessels, formation of pannus, destruction of cartilage and bone tissues and the like; in addition to joint diseases, inflammation or lesions can occur in the respiratory system, liver, kidney, heart and other parts, and clinically common therapeutic drugs mainly comprise the following types: non-steroidal anti-inflammatory drugs, antirheumatic drugs, biological agents, glucocorticoids, etc., have various side effects including gastrointestinal reactions, hepatorenal toxicity, etc. after long-term use; therefore, development or discovery of a drug which has little toxic and side effects and can prevent and treat the rheumatoid arthritis is of great importance.
Based on the components, characteristics and functions of the qinggong peach pill, the invention explores the prevention and treatment effects of the qinggong peach pill on the diseases through a collagen-adjuvant induced rheumatoid arthritis model of the autoimmune diseases of rats, lays a foundation for the research on the prevention and treatment effects of the qinggong peach pill on the autoimmune diseases, and provides clues and basis for the application of the qinggong peach pill in the aspect.
Disclosure of Invention
In view of the above, the invention aims to provide an application of a qinggong birthday peach pill in preparing an autoimmune regulation medicament so as to find out a medicament which has small toxic and side effects and can prevent and treat rheumatoid arthritis.
In order to achieve the above purpose, the technical scheme of the invention is realized as follows:
application of QINGGONGSHOUTAO pill in preparing medicine for regulating autoimmune function is provided.
Furthermore, the qinggonggao pill reduces the symptoms of rheumatoid arthritis by reducing the expression level of TNF-alpha, IFN-gamma, IL-6 and IL-17a in blood plasma.
Further, the administration dosage of the qinggong shou peach pill is 0.9-3.6 g/kg.
Further, the administration dosage of the qinggong shou peach pill is 1.8-3.6 g/kg.
Compared with the prior art, the invention has the following advantages:
after administration of the palace-clearing life peach pills with the dosage of 1.8g/kg and 3.6g/kg, the collagen-adjuvant-induced rat foot swelling area, arthritis score and the expression quantity of TNF-alpha, IFN-gamma, IL-6 and IL-17a in rat plasma are reduced, the symptoms of the rat rheumatoid arthritis are effectively relieved, and the good drug effect is exerted. The qinggonggao pill can participate in the treatment of the rheumatoid arthritis by regulating the immune response of the organism, thereby playing the role of the drug effect on autoimmune diseases; the research simultaneously provides clues and scientific basis for the fields of prevention, treatment and the like of autoimmune diseases.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention. In the drawings:
FIG. 1 is a graph of area of rat foot swelling;
FIG. 2 is a graph of a score for joint swelling in rats;
fig. 3 is a bar graph of inflammatory factor content in rat plasma, where C: normal control group; m: model control group; l: leflunomide control group; QL: low dose group of qinggong birthday peach pills; QM: dose group in qinggong shou peach pill; QH: high dosage of QINGGONGSHOUTAO pill.
Detailed Description
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
The invention will be described in detail below with reference to the drawings in connection with embodiments.
Autoimmune diseases have complex pathogenesis and various disease types, and in the research and development of new drugs, there are a plurality of models for evaluating the autoimmune regulation effect of drugs, wherein a collagen-induced rat rheumatoid arthritis model relates to two parts of cellular immunity and humoral immunity, and is one of representative models for evaluating the effect of the drugs; therefore, the rat rheumatoid arthritis model is selected in the study, the autoimmune regulation pharmacological action of the qinggong shou peach pill is further studied, and the humoral immunity and cell immunity regulation mechanisms are integrated, so that the Qinggong shou peach pill has representativeness and innovativeness.
1. Experimental materials
1.1 laboratory animals
SD rats: SPF grade, male, weight 180-220g, purchased from St Bei Fu (Beijing) Biotechnology Co., ltd., license number: SCXK (Beijing) 2021-0011.
Tianjin JinYao group Limited animal experiment center (Barrier Environment), license number: SCXK (Beijing) 2021-0011.
1.2 test article
Palace-cleaning birthday peach pill: lot number 8980021, available from Tianjin New pharmaceutical company Limited of the scientific research management department of Tianjin New pharmaceutical company, darentang pharmaceutical factory;
1.3 drugs and Agents
Leflunomide: specification 10mg, lot number 210410, available from the company of the pharmaceutical Co., ltd;
freund's complete adjuvant: specification 10mL, lot number 1003312942, purchased from Sigma;
chicken type II collagen: specification 2 mg/mL. Times.5 mL, available from Chondrex;
the rat IL-6ELISA kit, lot number Mar 2022, was purchased from Rui Xin Biotechnology Co., ltd,
rat IFN-gamma ELISA kit, lot number 20220111, purchased from Nanjing built Biotechnology Co., ltd;
rat IL-17a ELISA kit, lot 2101012434, rat TNF- α ELISA kit, lot GR3401548-1, all purchased from Abcam;
1.4 Main instruments
The vernier caliper is purchased from the market;
high-speed refrigerated centrifuge: s8TR, siemens Feishmania technology Co., ltd;
precision electronic balance: ME303E/02, metrele-Tolyduo instruments (Shanghai) Inc.;
full-wavelength multifunctional enzyme-labeled instrument: spectraMax i3x, molecular devices.
2. Experimental method
2.1 preparation of modeling pharmaceutical formulations
The chicken type II collagen and Freund's complete adjuvant are mixed in equal volume, blown in ice water bath to emulsify the solution thoroughly to form stable emulsifier, and the concentration of collagen in the final solution is 1.0mg/kg.
2.2 grouping, modeling and administration
Randomly dividing 60 rats into a normal control group, and building a module, wherein the normal control group comprises 5 rats and 55 rats; modular rats were injected intradermally with about 0.2mL collagen-adjuvant milk into the left hind pawDissolving liquid, molding for 21 days, measuring the foot swelling area of the rat, and selecting the foot swelling area to be larger than 90mm 2 The rats of (1) are randomly grouped according to the area of the swelling of feet, and are divided into 5 groups, wherein 10 rats in each group are respectively a model control group, a leflunomide group and a low, medium and high dose group of qinggong birthday peach pills, and each administration group is continuously orally administered for 14 days; the rats of each group are respectively subjected to gastric administration, the administration volume is 10mL/kg, the administration dosage of leflunomide is 5mg/kg, the administration dosage of the Qinggong birthday peach pill of the high, medium and low dosage groups is 3.6g/kg, 1.8g/kg and 0.9g/kg (which are 2 times, 1 time and 0.5 time of the clinical equivalent dosage), and the normal control group rats are subjected to gastric administration with drinking water of the same volume for 14 days.
2.3 foot swelling area measurement
The width and thickness of the left hind foot and contralateral foot of each group of rats are measured by using a vernier caliper, and the final foot swelling area is obtained by multiplying the two.
2.4 joint swelling severity score
The joint swelling degree of each foot of the rat was scored, and the final severity was obtained by summing the four foot scores, the scoring method being shown in table 1:
table 1 joint swelling severity score criteria
2.5 inflammatory factor determination
After the last administration for 30min, anesthetizing the rat, taking blood from abdominal aorta, collecting blood with procoagulant tube and EDTA anticoagulant tube, centrifuging at 3000rpm at room temperature for 10min, collecting serum and plasma, preserving at low temperature, and determining inflammatory factors in plasma;
according to the instruction book of the rat inflammatory factor ELISA measuring kit, inflammatory factors in blood plasma and blood serum are measured, the OD value of a sample is measured by a full-wavelength enzyme-labeled instrument, and finally, the concentration of the sample is calculated by using related software.
2.5 statistical methods
ˉ
Statistical analysis was performed with SPSS software, and the data were expressed as x+ -SD, with One-Way ANOVA for group comparisons, LSD for variance, dunnett-t for variance, and P <0.05 difference were statistically significant.
3. Experimental results
3.1 Effect of palace-clearing birthday peach pills on toe swelling in rats with rheumatoid arthritis
By measuring the area of foot swelling in rats during dosing, the area of foot swelling was increased in each group of rats before and after dosing compared to the normal control group, with a statistical difference (p < 0.01); during dosing, the area of foot swelling in the model control rats continues to increase; the areas of swelling of the feet of rats in the leflunomide group and the qinggonggao pill are reduced to different degrees during the administration period compared with the model control group, and the leflunomide group and the qinggonggao pill have the most obvious reduction (p < 0.05) in the high-dose (1.8 and 3.6 g/kg) group from the 8 th day to the 14 th day and have statistical significance compared with the model control group. The areas of the foot swelling of rats in the group of the Qinggong shou peach pills (1.8 g/kg) and the high dose (3.6 g/kg) reach the highest value, the areas of the foot swelling of rats continuously decrease from the 8 th day to the 14 th day, the areas of the foot swelling of rats in the group of each dose of the Qinggong shou peach pills are obviously reduced on the 14 th day, and compared with the model control group, the areas of the foot swelling of rats in the group of each dose of the Qinggong shou peach pills have obvious differences (p is less than 0.05), and the obvious differences are shown in tables 2.1 and 2.2.
TABLE 2.1 area of toe swelling for rats in each group [ (]Units: mm (mm) 2 )
P <0.01, compared to the normal control group; * P <0.05, < p <0.05, compared to model control group.
TABLE 2.2 inhibition of palace-clearing birthday peach pills on foot swelling in rats of each group%
3.2 Effect of palace-clearing birthday peach pills on joint swelling score of gouty arthritis-like rats
The scoring result shows that: the first day of dosing, except for the normal control group, the joint scores of the rats in each group were close, and the differences were not statistically significant; after administration, the scores of the groups gradually decrease, and on the 5 th day after administration, the scores of the leflunomide group, the qinggong shou peach pill and the high-dose group (1.8 g/kg and 3.6 g/kg) are reduced, but compared with the model control group, the scores of the leflunomide group, the qinggong shou peach pill and the high-dose group are not statistically different; the leflunomide group and the medium and high dose (1.8, 3.6 g/kg) groups showed a significant decrease in scores (p < 0.05) from day 8 to day 14 compared to the model control group, and were statistically significant, as shown in tables 3.1,3.2.
Table 3.1 joint swelling score for rats in each group
* P <0.05, < p <0.01, compared to model control group.
TABLE 3.2 inhibition of palace-clearing birthday peach pills on foot joint swelling score of rats of each group%
3.3 Effect of palace-clearing birthday peach pills on inflammatory factors in plasma of gouty arthritis-like rats
The Elisa assay showed that: the expression level of IL-6 in the plasma of the model control group is obviously increased (p < 0.01) compared with that of the normal control group; compared with the model control group, the expression level of IL-6 in the rat plasma of each dose group of the leflunomide group and the Qinggong birthday peach pill is reduced, wherein the function of the leflunomide group and the Qinggong birthday peach pill in the high dose (1.8 and 3.6 g/kg) group is most obvious (p < 0.01), and the significant difference exists. Compared with the normal control group, the expression level of IL-17a in the plasma of the model control group is obviously increased (p < 0.01) and has obvious difference; however, the expression of IL-17a was reduced to a different extent in each of the administration groups compared to the model control group, but there was no statistical difference. Compared with the normal control group, the model control group has obvious difference that the expression quantity of TNF-alpha in blood plasma is obviously increased (p < 0.01); compared with the model control group, the expression level of TNF-alpha in the blood plasma of rats in the leflunomide group and the Qinggong shou peach pill group is reduced (p is less than 0.01), and the obvious difference exists; the expression level of IFN-gamma in the rat plasma of the model control group is obviously increased, and compared with the normal control group, the IFN-gamma has obvious difference (p < 0.01); after administration, IFN-gamma of each dose group of the leflunomide group and the Qinggong birthday peach pill is reduced to different degrees, wherein the effect of the Qinggong birthday peach pill of the medium dose (1.8 g/kg) group is most obvious (p < 0.01), and the results are shown in tables 4.1 and 4.2.
TABLE 4.1 levels of individual inflammatory factors in plasma of groups of rats
P <0.05, #: p <0.05, compared to the normal control group; * P <0.05, compared to model control group.
TABLE 4.2 inhibition of Qinggong Shoupeach pills on inflammatory factors of rats of each group%
Conclusion 4
After administration of the palace-clearing life peach pills with the dosage of 1.8g/kg and 3.6g/kg, the collagen-adjuvant-induced rat foot swelling area, arthritis score and the expression quantity of TNF-alpha, IFN-gamma, IL-6 and IL-17a in rat plasma are reduced, the symptoms of the rat rheumatoid arthritis are effectively relieved, and the good drug effect is exerted. The study showed that: the qinggonggao pill can participate in the treatment of the rheumatoid arthritis by regulating the immune response of the organism, thereby playing the role of the drug effect on autoimmune diseases; the research simultaneously provides clues and scientific basis for the fields of prevention, treatment and the like of autoimmune diseases.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.
Claims (1)
1. Application of QINGGONGSHOUTAO pill in preparing medicine for treating rheumatoid arthritis is provided.
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| CN202211465289.5A CN115887593B (en) | 2022-11-22 | 2022-11-22 | Application of qinggongshou peach pill in preparing medicine for regulating autoimmune property |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109100431A (en) * | 2017-12-27 | 2018-12-28 | 天津中新药业集团股份有限公司达仁堂制药厂 | A kind of effective substance research method of palace of the Qing Dynasty Shoutao pills |
| CN109498768A (en) * | 2017-12-27 | 2019-03-22 | 天津中新药业集团股份有限公司达仁堂制药厂 | Application of the palace of the Qing Dynasty Shoutao pills in preparation treatment improvement decrease of memory, amnesia, cognitive disorder drug |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109100431A (en) * | 2017-12-27 | 2018-12-28 | 天津中新药业集团股份有限公司达仁堂制药厂 | A kind of effective substance research method of palace of the Qing Dynasty Shoutao pills |
| CN109498768A (en) * | 2017-12-27 | 2019-03-22 | 天津中新药业集团股份有限公司达仁堂制药厂 | Application of the palace of the Qing Dynasty Shoutao pills in preparation treatment improvement decrease of memory, amnesia, cognitive disorder drug |
Non-Patent Citations (3)
| Title |
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| 房定亚分期论治痛风性关节炎浅谈;韩淑花, 等;《中国中医基础医学杂志》;第24卷(第03期);第320-321页 * |
| 超氧化物歧化酶临床药理研究进展;冯高闳;《江西医学院学报》;第33卷(第04期);第103页左栏第2段、右栏第3段；第105页右栏第1段 * |
| 陈可冀治学学术思想临床经验暨清宫医药老年医学研究成就初步探讨(续);李永强, 等;《新疆中医药》;第27卷(第02期);第78-84页 * |
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