A plaster for treating granulomatous lobular mastitis and a preparation method thereof
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- THE FIRST HOSPITAL OF HUNAN UNIV OF CHINESE MEDICINE (CLINICAL RES INST OF TRADITIONAL CHINESE MEDICINE)
- Filing Date
- 2025-02-10
- Publication Date
- 2026-06-02
Smart Images
Figure CN119770603B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of traditional Chinese medicine technology, and more specifically, to a plaster for treating granulomatous lobular mastitis and its preparation method. Background Technology
[0002] Granulomatous lobular mastitis (GLM), also known as idiopathic granulomatous mastitis, is a rare inflammatory breast disease of unknown etiology. It is characterized by non-caseating granulomatous inflammation of the lobules, which may be accompanied by infiltration of lymphocytes, neutrophils, plasma cells, monocytes, and eosinophils. It was first described by Kessler E and Wolloch Y in 1972. GLM is not common clinically, especially in Europe and America, but its incidence has been increasing in recent years. It is most common in women of childbearing age, particularly multiparous women with a history of breastfeeding. It can also occur in breastfeeding women, pregnant women, or women without a history of childbirth, and in a small number of men. The clinical manifestations of GLM are varied and diverse. In the early stages of the disease, a lump often appears in the breast, with or without significant pain. The lump is relatively firm and poorly demarcated from the surrounding tissue. In some cases, the condition progresses rapidly, and the lump may affect the entire breast. In later stages, suppuration occurs in some cases, and in severe cases, multiple sinuses or fistulas appear, with persistent and difficult-to-heal pus discharge. Some patients may also experience systemic symptoms such as erythema nodosum on the limbs, fever, and cough. Traditional Chinese medicine does not have a clear record of this disease, but based on its characteristics and clinical manifestations, it can be classified under the categories of "mastitis" or "mast fistula".
[0003] Currently, Western medical treatments for granulomatous lobular mastitis (GLM) are diverse and still controversial, broadly categorized into surgical and non-surgical treatments. Traditional Chinese medicine (TCM) advocates a holistic approach, emphasizing combined internal and external treatments. Internal treatments, based on staged treatment, appropriately incorporate unique TCM external therapies, achieving relatively good results. Although this disease is benign, its local invasiveness can cause long-term pain and psychological distress for patients. Therefore, the inventors propose a plaster for treating granulomatous lobular mastitis and its preparation method. Summary of the Invention
[0004] The purpose of this invention is to provide a plaster for treating granulomatous lobular mastitis and its preparation method, thereby achieving the treatment of granulomatous lobular mastitis.
[0005] The above-mentioned technical objective of the present invention is achieved through the following technical solution:
[0006] The first aspect of this invention provides a plaster for treating granulomatous lobular mastitis. By weight, the raw materials of the plaster for treating granulomatous lobular mastitis are composed of the following components: 13-16 parts of rhubarb, 8-12 parts of phellodendron bark, 8-12 parts of scutellaria root, 13-16 parts of burnet root, 8-12 parts of gardenia fruit, 18-22 parts of turmeric, and 13-16 parts of lithospermum root.
[0007] In conjunction with the first aspect, the present invention is further configured as follows: 15 parts rhubarb, 10 parts phellodendron bark, 10 parts scutellaria root, 15 parts burnet root, 10 parts gardenia fruit, 20 parts turmeric, and 15 parts gromwell root.
[0008] Rhubarb: Bitter, cold. It enters the spleen, stomach, large intestine, liver, and pericardium meridians. It has purgative and detoxifying properties, clears heat and fire, cools the blood and detoxifies, removes blood stasis and promotes menstruation, and promotes diuresis and relieves jaundice. It is used for constipation due to excess heat, hematemesis and epistaxis due to blood heat, red eyes and swollen throat, carbuncles and boils, intestinal abscesses and abdominal pain, amenorrhea due to blood stasis, postpartum blood stasis, traumatic injuries, damp-heat dysentery, jaundice with dark urine, urinary tract infections, and edema; externally, it is used to treat burns and scalds.
[0009] Phellodendron bark: Bitter, cold. Enters the kidney and bladder meridians. Clears heat and dries dampness, drains fire and eliminates steaming heat, detoxifies and heals sores. Used for damp-heat diarrhea, jaundice with dark urine, leukorrhea and vulvar itching, painful urination due to heat, beriberi with weakness and paralysis, steaming bone fever, night sweats, seminal emission, sores and carbuncles, eczema and damp sores. Salt-processed Phellodendron bark nourishes yin and reduces fire. Used for yin deficiency with excessive fire, night sweats and steaming bone fever.
[0010] Scutellaria baicalensis: Bitter, cold. It enters the lung, gallbladder, spleen, large intestine, and small intestine meridians. It clears heat and dries dampness, drains fire and detoxifies, stops bleeding, and calms the fetus. It is used for damp-heat syndrome, summer-heat syndrome, chest tightness and nausea, damp-heat fullness, diarrhea, jaundice, lung-heat cough, high fever and thirst, blood-heat vomiting and nosebleeds, carbuncles and boils, and threatened abortion.
[0011] Sanguisorba officinalis: Bitter, sour, and astringent; slightly cold. It enters the liver and large intestine meridians. It cools the blood and stops bleeding, detoxifies and astringes sores. Used for hematochezia, hemorrhoidal bleeding, bloody dysentery, metrorrhagia, burns, carbuncles, and boils.
[0012] Gardenia: Bitter, cold. Enters the Heart, Lung, and Triple Energizer meridians. Clears heat and relieves irritability, promotes diuresis and eliminates dampness, cools the blood and detoxifies; externally used to reduce swelling and relieve pain. Used for feverish irritability, damp-heat jaundice, painful urination, hematemesis and epistaxis due to blood heat, red and swollen eyes, and carbuncles due to fire toxins; externally used to treat sprains and contusions.
[0013] Turmeric: pungent, bitter, and warm. It enters the spleen and liver meridians. It breaks up blood stasis, promotes qi circulation, unblocks meridians, and relieves pain. It is used for stabbing pain in the chest and hypochondrium, chest pain, dysmenorrhea, amenorrhea, abdominal masses, rheumatic shoulder and arm pain, and swelling and pain from falls.
[0014] Purple gromwell root: Sweet, salty, and cold in nature. It enters the heart and liver meridians. It cools the blood, invigorates blood circulation, detoxifies, and promotes rash eruption. It is used for blood heat and toxicity, purplish-black rashes, incomplete measles eruption, sores, eczema, and burns.
[0015] In this formula, rhubarb, phellodendron bark, and scutellaria are the chief herbs, working together to clear heat, purge fire, cool the blood, and detoxify. When heat and toxins accumulate and condense into phlegm, the phlegm and heat obstruct the flow of qi, causing stagnation of qi and blood. "Where there is no flow, there is pain." Turmeric can invigorate blood, unblock the meridians, and relieve pain, serving as the assistant herb. Sanguisorba officinalis and gardenia are bitter and cold in nature, assisting the chief herbs in enhancing their ability to cool the blood, purge fire, and detoxify. Lithospermum erythrorhizon is sweet, salty, and cold in nature, not only clearing heat, cooling the blood, and detoxifying, but also enhancing the ability to invigorate blood and reduce swelling when combined with turmeric. The entire formula can clear heat, purge fire, and detoxify, as well as invigorate blood and reduce swelling, ultimately allowing the swelling caused by yang syndrome to dissipate.
[0016] The second aspect of this invention also provides a method for preparing a plaster for treating granulomatous lobular mastitis. The method involves adding water to all the ingredients and decocting twice. In the first decoction, 9-11 times the amount of water as the ingredients are added, and the decoction is decocted for 1-3 hours. In the second decoction, 6-9 times the amount of water as the ingredients are added, and the decoction is decocted for 1-2 hours. The two decoctions are combined, filtered, and concentrated to obtain a plaster. The plaster is then applied, cut to size, and allowed to solidify.
[0017] In conjunction with the second aspect, the present invention is further configured such that: the first decoction time is 2 hours, and the second decoction time is 1.5 hours.
[0018] In conjunction with the second aspect, the present invention is further configured such that the concentration is carried out at 60°C to a relative density of 1.15 to 1.18.
[0019] A third aspect of the present invention also provides for the use of any of the above-prepared plasters in the preparation of a drug for treating granulomatous lobular mastitis.
[0020] In conjunction with the third aspect, the present invention is further configured such that: the drug is a drug that reduces the expression of IL6, CD86, TYROBP, ITGAX and regulates related pathways.
[0021] In summary, the present invention has the following beneficial effects:
[0022] 1. Through gene sequencing and bioinformatics analysis, and further verification by animal experiments, PTPRC, IL6, IL1B, IL10, CD86, FCGR3A, TYROBP, ITGAX, CXCL8, and SPI1 are potential genes for the pathogenesis of granulomatous lobular mastitis. Except for IL10, which is a downregulated gene, the other genes are upregulated genes.
[0023] 2. Animal experiments have shown that Ruyi Ointment is effective in treating granulomatous lobular mastitis and can reduce the breast inflammation index score and the degree of inflammatory cell infiltration.
[0024] 3. Ruyi Ointment exerts its therapeutic effect on granulomatous lobular mastitis by reducing the expression of IL6, CD86, TYROBP, and ITGAX and regulating related pathways. Attached Figure Description
[0025] Figure 1This is the GO and KEGG pathway analysis of differentially expressed genes in Experiment Example 1 of this invention (Figure A is a heatmap of differentially expressed genes, Control is the normal group, GLM is the granulomatous lobular mastitis group, red represents gene upregulation, and blue represents gene downregulation; Figure B is a volcano diagram of differentially expressed genes, red represents differentially upregulated genes, and blue represents differentially downregulated genes; Figure C is a GO analysis diagram of differentially expressed genes; Figure D is a KEGG pathway analysis diagram of differentially expressed genes).
[0026] Figure 2 This is the PPI network diagram of the first ten Hub genes in Experiment Example 1 of this invention;
[0027] Figure 3 This refers to the effect of Ruyi Ointment on the gross appearance of rat mammary glands in Experimental Example 2 of this invention;
[0028] Figure 4 The effect of Ruyi Ointment on the pathological characteristics of rat mammary tissue in Experimental Example 2 of this invention (HE, ×400);
[0029] Figure 5 This relates to the effect of Ruyi Ointment on the expression level of Hub gene protein in rat mammary tissue in Experiment Example 2 of this invention. Detailed Implementation
[0030] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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.
[0031] Example 1:
[0032] Take 15g of rhubarb, 10g of phellodendron bark, 10g of scutellaria root, 15g of burnet root, 10g of gardenia fruit, 20g of turmeric, and 15g of gromwell root, and decoct them twice with water. For the first decoction, add 10 times the amount of water as the herbs and decoct for 2 hours. For the second decoction, add 8 times the amount of water and decoct for 1.5 hours. Combine the two decoctions, filter, and concentrate to a relative density of 1.15-1.18 (60℃). After forming a paste, spread it on a plate, cut it to size, and let it solidify to shape.
[0033] Experiment Example 1: Gene Sequencing and Hub Gene Screening
[0034] 1.1 General Information
[0035] Ten patients who underwent breast biopsy at the Department of Breast Surgery, First Affiliated Hospital of Hunan University of Traditional Chinese Medicine, and were diagnosed with granulomatous lobular mastitis and fibroadenoma respectively by postoperative pathology, were included in this study. During the operation, surgical specimens of granulomatous lobular mastitis and normal breast tissue were collected. Written informed consent was obtained from all volunteers for the collection of all tissue specimens.
[0036] 1.2 Pathological sampling
[0037] Fresh surgical specimens were collected during the operation. The collected specimens were flash-frozen in liquid nitrogen and then stored in a -80°C freezer. All specimens were numbered for subsequent testing.
[0038] 1.3 Genetic Testing
[0039] 1.3.1 RNA Sample Detection
[0040] First, RNA was extracted from the samples using standard extraction methods, and then the RNA samples underwent strict quality control, mainly including the following three aspects: (1) Concentration quality control: the concentration and total amount were detected using a Qubit 4.0 instrument. (2) Fragment integrity quality control: the fragment integrity of the RNA samples was assessed using an Agilent 2100 analyzer. (3) Purity quality control: the purity of the samples was controlled using a NanoDrop spectrophotometer.
[0041] 1.3.2 mRNA Acquisition, Library Construction, and Quality Control
[0042] Since most eukaryotic mRNAs possess a polyA tail, Oligo(dT) magnetic beads can be used to enrich these polyA-tailed mRNAs. A metal ion solution treatment method was employed, with appropriate fragmentation temperatures and times selected based on the desired fragment size. Using the fragmented mRNA as a template, the first cDNA strand was synthesized in an M-MuLV reverse transcriptase system using random oligonucleotides as primers. After the first strand synthesis was complete, second-strand synthesis, end repair, and A-tail addition were performed. T4 DNA ligase was used to catalyze the ligation of the sticky ends of the double-stranded DNA to short adapters. The ligation products were purified using AMPureXP beads, and cDNA fragments of approximately 300-350 bp were selected, ensuring fragment concentration. PCR amplification was then performed, with sample index tags added. The PCR products were then purified using AMPureXP beads, ultimately yielding a library. After library construction, preliminary quantification was performed. After diluting the library, the distribution of library fragments was detected using an Agilent 2100 DNA1000kit. After passing quality control, the effective concentration of the library was accurately quantified by qRT-PCR to ensure library quality.
[0043] 1.3.4 Sequencing and Data Quality Control
[0044] The prepared DNB was loaded onto a patterned array and PE150 sequencing was performed using combined probe-anchor synthesis (cPAS). Sequencing primers and fluorescent probes were polymerized on DNA nanospheres, and then the light signals were acquired, read, and identified using a high-resolution imaging system to obtain individual base sequence information. The process was then repeated to obtain the sequence information for the next base. After high-throughput sequencing, the raw image data files were converted into raw sequencing sequences through base identification and analysis. To ensure the quality of information analysis, reads shorter than 35 bp after excision of adapter sequences and low-quality bases at both ends, reads with an N (meaning base information cannot be determined) ratio greater than 10% of the total bases, and reads where low-quality bases (quality value Q <= 5) accounted for more than 40% of the total read length were removed using the FastP1 software. This resulted in high-quality clean data, which was then analyzed for base count, sequence count, and other statistical parameters.
[0045] 1.4 Differentially expressed gene analysis
[0046] After completing the quantitative analysis of genes, the expression levels of all samples were merged into an expression matrix, and differential expression significance analysis was performed at the gene or transcript level to identify functionally differentially expressed genes or transcripts related to sample groups. The DESeq2 software was used for differential significance analysis, with a corrected P-value ≤ 0.001 and |log2FC| ≥ 2.5 used as the criteria for significant difference.
[0047] 1.5 GO and KEGG pathway analysis of differentially expressed genes
[0048] First, all differentially expressed genes were mapped to relevant entries in the GO database (http: / / www.geneontology.org / ), and the number of differentially expressed genes in each entry was counted, followed by statistical tests. A p-value ≤ 0.05 was considered significant enrichment, identifying GO entries that were significantly enriched in differentially expressed genes compared to the entire genome background. KEGG is a major public database for pathways. Pathway significant enrichment analysis was performed on a KEGG pathway-by-pathway basis, using hypergeometric tests, with a p-value ≤ 0.05 as the threshold for significant enrichment, identifying pathways that were significantly enriched in differentially expressed genes compared to the entire genome background.
[0049] 1.6 PPI network construction and Hub gene identification
[0050] To evaluate potential protein - protein interaction relationships, the previously identified differentially expressed genes were mapped to a database for retrieving gene interactions (http: / / string - db.org / ) to construct a Protein - Protein Interaction (PPI) network. Subsequently, the PPI network was visually analyzed using Cytoscape software, and the top ten genes in the PPI network were selected as key genes (Hub genes) through the Degree algorithm. Hub genes, which play crucial roles in biological processes, often influence the regulation of other genes in related signaling pathways. In gene - level research, Hub genes are regarded as important target sites and research hotspots.
[0051] Experimental Example 2
[0052] Experimental materials
[0053] 18 female SD rats (250 - 300 g), uniformly ordered from the Dongtang Experimental Animal Center of Hunan University of Chinese Medicine (Hunan Slack Jingda Experimental Animal Co., Ltd.), license number: SCXK(Xiang)2019 - 0004.
[0054] Drugs and reagents
[0055] The plaster prepared in Example 1
[0056] Table 1 - 1 Information table of drugs and reagents
[0057]
[0058]
[0059] Preparation of homogenate
[0060] The frozen specimens of human granulomatous lobular mastitis tissue were taken out from the - 80°C refrigerator and placed in a 37°C water bath for thawing. After thawing, the mammary gland tissue in the cryopreservation tube was taken out and weighed with a precision electronic balance. The tissue was cut into pieces with scissors until there were no obvious large particles, and then mixed with 0.9% NaCl solution at a ratio of 1:3 and placed in a 15 - ml centrifuge tube. The sample homogenization treatment system was used for cell crushing treatment for about 5 minutes to release the proteins in the cytoplasm into the solution. An appropriate amount of the supernatant was taken from the homogenate after cell crushing treatment and placed in a sterile centrifuge tube. Freund's complete adjuvant was added to the supernatant at a volume ratio of 1:1, and the mixture was repeatedly aspirated and mixed with a Pasteur pipette to prepare an oil - in - water emulsion suspension, which was stored at 4°C in the refrigerator for later use.
[0061] Experimental method
[0062] 2.1 Grouping and Modeling
[0063] Eighteen rats were numbered 1-18 and randomly divided into three groups using a random number table: a control group, a model group, and an experimental group (Ruyi Ointment group), with six rats in each group. The rats were allowed to acclimatize for three days. The experimental and model groups were anesthetized by intraperitoneal injection of a prepared 2% pentobarbital solution at a dose of 0.2 mL / 100 g. After successful anesthesia, hair was removed from the third and fourth pairs of mammary glands and surrounding areas using depilatory cream. The nipples and surrounding skin were disinfected with iodine. 0.2 ml of a pre-prepared suspension was drawn using a 1 ml syringe and injected into the third and fourth pairs of mammary glands of the rats, completing the artificial implantation process. Simple pressure was applied to stop bleeding after needle removal. No treatment was given to the normal group rats.
[0064] Validation of the granulomatous lobular mastitis model: Through preliminary experimental validation, before the experiment, 5 SD rats were used to establish the model using the above method. The general condition was observed and recorded. After obvious local lumps appeared, the model rats were sacrificed and samples were taken for pathological observation such as HE staining. The presence of granulomatous lesions and the general condition of local redness and swelling (positive evidence model) indicated that the model was successfully established.
[0065] 2.2 Administration
[0066] After successful modeling, the experimental group was given a 4cm×3cm Ruyi Ointment applied to the 3rd and 4th pairs of breasts, while the control group (normal group and model group) received no treatment. After administration, the breasts were bandaged and fixed. The administration frequency was once a day for 14 consecutive days.
[0067] 2.3 General Observation
[0068] After modeling and drug administration, rats in each group were routinely maintained and fed. The performance of rats in each group was observed daily, and changes in the animals were recorded, including rat condition, presence of mammary gland redness and swelling, abscess formation, and skin color. Based on the rats' performance, a mammary gland inflammation index score was assigned.
[0069] Table 1-2 Breast Inflammation Index Scoring Table
[0070]
[0071]
[0072] 2.4 Collection of materials
[0073] All rats were sacrificed on day 14 after drug administration, and the third and fourth pairs of mammary gland tissue were excised. One part was rinsed with physiological saline and fixed in 4% paraformaldehyde solution, while the other part was stored in a -80°C freezer for later RT-qPCR and Western blot experiments.
[0074] 2.2.2.5 Observation Indicators and Indicator Testing
[0075] (1) Observe the local and overall mammary gland manifestations of rats in each group, and score them according to the breast inflammation index scoring table set above. The final breast inflammation index is the sum of the scores of each item.
[0076] (2) Breast tissue was routinely embedded in paraffin and sectioned to a thickness of 4 μm. Intact breast tissue was selected from the sections, dewaxed, stained with hematoxylin and eosin (HE), and finally dehydrated and mounted. The morphological characteristics of the breast tissue and the infiltration of inflammatory cells were observed under an optical microscope.
[0077] (3) Western blot assay was used to detect the expression levels of target gene proteins in the above-mentioned gene sequencing in each tissue sample: A suitable amount of rat mammary gland tissue was cut, lysed with RIPA lysis buffer, and centrifuged to obtain protein supernatant. Protein denaturation was performed simultaneously with gel preparation. After gel loading and electrophoresis, filter paper and NC membrane were first placed together in transfer buffer for transfer treatment. Then, the membrane was soaked in 5% skim milk powder prepared with PBST and allowed to stand at room temperature for 90 minutes. Next, the primary antibody was diluted with PBST at a certain ratio and incubated with the membrane at 4°C overnight. The next day, it was allowed to stand at room temperature for 30 minutes. Then, the HRP-labeled secondary antibody was diluted with PBST and incubated with the membrane at room temperature for 90 minutes. Finally, the membrane was incubated with ECL chemiluminescence solution for 1 minute, then wrapped with plastic film and placed in a gel imaging system for imaging.
[0078] (4) Real-time PCR detection of potential target gene expression: Total RNA was extracted from rat mammary tissue using the Trizol method. The concentration and purity of RNA were measured using a UV spectrophotometer. Total mRNA from the tissue was used as a template for reverse transcription to generate cDNA. SYBR fluorescent dye and upstream and downstream primers were added for real-time quantitative PCR experiments (3 wells per sample per indicator, total volume 30 μL, 10 μL per well). Finally, a quantitative PCR amplification program was performed. The target gene sequence was retrieved from the NCBI database. Primers were designed using Primer5 software and synthesized by Beijing Qingke. Specific primer information is shown in the table below.
[0079] Table 1-4 Primer sequences and primer lengths
[0080]
[0081] Note: The CXCL8 primer sequence was not found in NCBI, therefore Real-time PCR was not performed.
[0082] 2.3 Statistical Methods
[0083] SPSS 25.0 software was used for statistical analysis, and GraphPad Prism 9.0 was used to generate statistical charts. Quantitative data were presented in [the format of the chart]. This indicates that when the data conforms to normality and homogeneity of variance, one-way ANOVA is used to compare differences between groups; when the data does not conform to normality and / or homogeneity of variance, the Kruskal-Wallis H test is used to compare differences between groups. P < 0.05 is set as statistically significant.
[0084] 3 Experimental Results
[0085] 3.1 GO and KEGG pathway analysis of differentially expressed genes
[0086] Using P-value ≤ 0.001 and |log2FC| ≥ 2.5 as the criteria for significant difference, the sequencing data were analyzed using DESeq2 software. A total of 1210 differentially expressed genes (DEGs) were screened, including 1095 upregulated genes and 115 downregulated genes. The DEGs of this dataset were visualized. Figure 1 A, B). GO and KEGG pathway analysis was performed on the identified differentially expressed genes. Figure 1C, D) GO analysis showed that biological process (BP) was mainly enriched in leukocyte-mediated immunity, positive regulation of cell activation, and positive regulation of leukocyte activation; cellular component (CC) was mainly enriched in the external side of the plasma membrane, plasma membrane receptor complex, and immunoglobulin complex; and molecular function (MF) was mainly enriched in antigen binding, immune receptor activity, and cytokine activity. KEGG pathway analysis showed that DEGs were enriched in cytokine-cytokine receptor interaction, chemokine signaling pathway, and viral protein interaction with cytokine and cytokine receptor.
[0087] Construction of differentially expressed gene PPI network and identification of Hub gene
[0088] The 1210 differentially expressed genes (DEGs) selected were imported into the string website to obtain the PPI network. The top ten genes (Hub genes) were selected using the Degree algorithm, in the following order: PTPRC, IL6, IL1B, IL10, CD86, FCGR3A, TYROBP, ITGAX, CXCL8, and SPI1. Figure 2 ).
[0089] Ruyi ointment reduced the breast inflammation index in GLM model rats.
[0090] Depend on Figure 3As observed, the model group rats showed localized redness and swelling in the mammary glands, with obvious mass formation, and some accompanied by abscess formation. Compared with the model group, the experimental group rats showed reduced redness and swelling in the mammary glands, smaller masses, and no abscess formation. Table 2-1 shows that compared with the normal group, the model group rats had an increased mammary gland inflammation index (P < 0.05); compared with the model group, the experimental group rats treated with Ruyi ointment showed a decreased mammary gland inflammation index (P < 0.05).
[0091] Table 2-1 Effects of Ruyi Ointment on Mammary Inflammation Index in Rats
[0092]
[0093] Note: 1) Compared with the blank group, 2) Compared with the model group.
[0094] Ruyi ointment reduced the degree of inflammatory cell infiltration in the mammary tissue of GLM model rats.
[0095] In normal rat mammary tissue, the lobules and ducts were clearly defined, with relatively dense cell arrangement. Compared to the control group, the model group showed lesions involving the terminal duct lobules in the mammary tissue, with most lobules having blurred outlines and indistinct structures. Lymphocytes, plasma cells, and macrophages infiltrated the lesions, and granulomas formed by the aggregation of epithelioid cells were observed within many lobules. Compared to the model group, the experimental group showed reduced infiltration of inflammatory cells such as lymphocytes and plasma cells, and decreased the formation of inflammatory granulomas. Figure 4 ).
[0096] Ruyi ointment affects the mRNA expression of some Hub genes.
[0097] As shown in Table 2-2, compared with the blank group, the relative expression levels of PTPRC, IL6, IL1B, CD86, FCGR3A, TYROBP, ITGAX, and SPI1 mRNA in the mammary tissue of rats in the model group were significantly increased (P < 0.001), while the relative expression level of IL10 mRNA was significantly decreased (P < 0.001), showing statistically significant differences. Compared with the model group, the experimental group significantly downregulated the expression of PTPRC, IL6, IL1B, CD86, FCGR3A, TYROBP, ITGAX, and SPI1 mRNA (P < 0.01), while upregulating the expression of IL10 mRNA (P < 0.01).
[0098] Table 2-2 Effects of Ruyi Ointment on the relative expression level of Hub gene mRNA in rat mammary tissue (2 -ΔΔCt (X±SD, n=3)
[0099]
[0100] Note: Compared with the blank group, 1) P < 0.001; compared with the model group, 2) P < 0.01.
[0101] 3.6 Ruyi Ointment Affects Protein Expression of Some Hub Genes
[0102] Compared with the control group, the expression of PTPRC, IL6, IL1B, CD86, FCGR3A, TYROBP, ITGAX, CXCL8, and SPI1 proteins in the mammary tissue of rats in the model group was significantly increased (P < 0.01), while the protein expression of IL10 was decreased compared with the control group (P < 0.05). Compared with the model group, the protein expression of IL6, CD86, TYROBP, and ITGAX was decreased in the experimental group (P < 0.05), while the protein expression of the remaining Hub genes showed no significant difference. Figure 5 (Table 2-3)
[0103] Table 2-3 Effects of Ruyi Ointment on Hub gene protein expression in rat mammary tissue (X±SD, n=3)
[0104]
[0105] Note: Compared with the blank group, 1) P < 0.01, 2) P < 0.05; compared with the model group, 3) P < 0.05
[0106] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A plaster for treating granulomatous lobular mastitis, characterized in that: The raw materials of the plaster for treating granulomatous lobular mastitis, by weight, consist of the following components: 13-16 parts rhubarb, 8-12 parts phellodendron bark, 8-12 parts scutellaria root, 13-16 parts burnet root, 8-12 parts gardenia fruit, 18-22 parts turmeric, and 13-16 parts gromwell root.
2. The plaster for treating granulomatous lobular mastitis according to claim 1, characterized in that: 15 parts rhubarb, 10 parts phellodendron bark, 10 parts scutellaria root, 15 parts burnet root, 10 parts gardenia fruit, 20 parts turmeric, and 15 parts gromwell root.
3. A method for preparing a plaster for treating granulomatous lobular mastitis as described in any one of claims 1-2, characterized in that: All the ingredients in the formula are decocted twice with water. For the first decoction, add 9-11 times the amount of water as the ingredients and decoct for 1-3 hours. For the second decoction, add 6-9 times the amount of water as the ingredients and decoct for 1-2 hours. Combine the two decoctions, filter, and concentrate to obtain a paste. Apply the paste, cut it to size, and let it solidify.
4. The method for preparing a plaster for treating granulomatous lobular mastitis according to claim 3, characterized in that: The first decoction time is 2 hours, and the second decoction time is 1.5 hours.
5. The method for preparing a plaster for treating granulomatous lobular mastitis according to claim 3, characterized in that: The concentration is carried out at 60°C to a relative density of 1.15 to 1.
18.
6. The use of the plaster according to any one of claims 1-2 or the plaster prepared by the preparation method according to any one of claims 3-5 in the preparation of a drug for treating granulomatous lobular mastitis.