Traditional Chinese medicine composition for treating heart yin deficiency and myocardial fibrosis and application of traditional Chinese medicine composition

Abstract

The invention belongs to the field of biological medicine, and particularly relates to a traditional Chinese medicine composition for treating heart yin deficiency and myocardial fibrosis and application thereof. The traditional Chinese medicine composition is prepared from the following raw medicinal materials in parts by weight: 5-13 parts of lily, 5-13 parts of radix ophiopogonis, 3-9 parts of ginseng, 5-13 parts of radix scrophulariae, 3-9 parts of semen boitae and 1-5 parts of schisandra chinensis. The invention also provides a preparation method of the composition. The composition has the effects of reinforcing qi, nourishing yin, calming the heart and tranquilizing the mind, and can be used for preparing medicines for treating heart-yin deficiency and myocardial fibrosis or improving heart-yin deficiency symptoms. Experimental results show that the traditional Chinese medicine composition can significantly improve the heart function of a chronic heart failure model mouse, inhibit myocardial cell hypertrophy and collagen deposition, and down-regulate fibrosis-related gene expression. Clinical observation shows that the traditional Chinese medicine can effectively relieve the symptoms of palpitation, insomnia, dysphoria with smothery sensation and the like of patients with heart yin deficiency. The traditional Chinese medicine composition is clear in formula and controllable in quality, and a novel safe and effective traditional Chinese medicine choice is provided for treating heart yin deficiency related myocardial fibrosis.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to a traditional Chinese medicine composition for treating myocardial fibrosis due to deficiency of heart yin and its application. Background Technology

[0002] Heart Yin Deficiency Syndrome is a common syndrome in Traditional Chinese Medicine (TCM), referring to the depletion of Yin fluid in the heart, leading to internal heat. Clinical manifestations include palpitations, insomnia, five-center heat (heat in the palms, soles, and chest), dry mouth and throat, etc., and it is commonly seen in the protracted stage of cardiovascular diseases such as chronic heart failure. Modern medicine believes that the continuous activation of chronic inflammation and fibrosis signals during the progression of heart failure is a key pathological link leading to myocardial interstitial fibrosis and poor ventricular remodeling. From the perspective of "syndrome-disease integration," the state of "internal disturbance of deficiency heat and malnourishment of the collaterals" in Heart Yin Deficiency is intrinsically related to this pathological process.

[0003] Currently, the treatment of heart yin deficiency syndrome often employs classic formulas such as Tianwang Buxin Dan and Zhigancao Tang to nourish yin and calm the mind, or uses Western medicines such as sleeping pills and beta-blockers for symptomatic treatment. However, classic formulas are complex, difficult to standardize in terms of quality, and generally lack targeted design for myocardial fibrosis, a structural lesion; while Western medicines focus on short-term symptom control and are difficult to regulate the core pathogenesis of "heart yin deficiency and internal disturbance of deficiency heat," thus having limited effect on reversing myocardial fibrosis and adverse remodeling, and long-term use carries the risk of dependence and adverse reactions.

[0004] Therefore, there is an urgent need to develop a traditional Chinese medicine compound that is simple in formulation, has controllable quality, can improve the core symptoms of heart yin deficiency, and can effectively intervene in the process of myocardial fibrosis, so as to meet the clinical needs for comprehensive treatment of heart diseases related to heart yin deficiency. Summary of the Invention

[0005] This invention addresses the common problems in existing drug treatments for myocardial fibrosis related to heart yin deficiency, such as "complex formulations, difficulty in quality control, and difficulty in simultaneously improving symptoms and intervening in structural intervention." It provides a traditional Chinese medicine composition for treating myocardial fibrosis caused by heart yin deficiency, along with its preparation method and application. Composed of six herbs—lily bulb, ophiopogon japonicus, ginseng, scrophularia ningpoensis, arborvitae seed, and schisandra chinensis—in a specific ratio, this composition exerts the effects of tonifying qi and nourishing yin, clearing heat and calming the mind. While improving symptoms of heart yin deficiency, it also inhibits the progression of myocardial fibrosis and improves cardiac function, achieving an integrated treatment that combines disease and syndrome.

[0006] To achieve the above objectives, the present invention adopts the following technical solution.

[0007] The present invention provides a traditional Chinese medicine composition for treating myocardial fibrosis caused by heart yin deficiency, which is composed of six raw medicinal materials, namely lily, ophiopogon root, ginseng, scrophularia root, semen platycladi and schisandra chinensis, and is named "Baimai Ningxin Formula"; specifically, the traditional Chinese medicine composition is composed of the following raw medicinal materials in parts by weight: 5-13 parts of lily, 5-13 parts of ophiopogon root, 3-9 parts of ginseng, 5-13 parts of scrophularia root, 3-9 parts of semen platycladi, and 1-5 parts of schisandra chinensis.

[0008] In some embodiments of the present invention, the traditional Chinese medicine composition is composed of the following raw medicinal materials in parts by weight: 7-11 parts of lily, 7-11 parts of ophiopogon root, 5-7 parts of ginseng, 7-11 parts of scrophularia root, 5-7 parts of semen platycladi, and 2-4 parts of schisandra chinensis.

[0009] In some embodiments of the present invention, the traditional Chinese medicine composition is composed of the following raw medicinal materials in parts by weight: 9 parts of lily, 9 parts of ophiopogon root, 6 parts of ginseng, 9 parts of scrophularia root, 6 parts of semen platycladi, and 3 parts of schisandra chinensis.

[0010] The present invention also discloses a preparation method of the traditional Chinese medicine composition, including: taking the raw medicinal materials of the traditional Chinese medicine composition according to the formula, extracting with water, and collecting the extractive solution to obtain the product.

[0011] The present invention also discloses a drug for treating myocardial fibrosis caused by heart yin deficiency, and the drug uses the traditional Chinese medicine composition as an active ingredient.

[0012] In some embodiments of the present invention, the drug is a decoction, granule, pill, capsule, tablet, powder or oral liquid.

[0013] In some embodiments of the present invention, the drug further includes other pharmaceutically acceptable excipients.

[0014] In some embodiments of the present invention, the other pharmaceutically acceptable excipients include: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, antiadhesives, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, clathrates, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, and release retardants.

[0015] In some embodiments of the present invention, the filler is selected from starch, sucrose, lactose, or microcrystalline cellulose; the binder is selected from cellulose derivatives, alginate, gelatin, or polyvinylpyrrolidone; the wetting agent is selected from water, glycerin, ethanol, methylcellulose, sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, or hydroxypropyl methylcellulose; the disintegrant is selected from sodium carboxymethyl starch, hydroxypropyl cellulose, cross-linked carboxymethyl cellulose, agar, calcium carbonate, or sodium bicarbonate; the absorption promoter is a quaternary ammonium compound; the surfactant is selected from hexadecyl alcohol or sodium dodecyl sulfate; the adsorbent carrier is selected from kaolin or soap clay; the lubricant is selected from talc, calcium and magnesium stearate, micronized silica gel, or polyethylene glycol; and the flavoring agent is selected from sucrose, simple syrup, aromatic syrup, glycerin, sorbitol, or mannitol.

[0016] In some embodiments of the present invention, the drug further includes a drug carrier.

[0017] In some embodiments of the present invention, the drug carrier includes microcapsules, microspheres, nanoparticles, and liposomes.

[0018] The present invention also discloses the application of the traditional Chinese medicine composition in the preparation of a drug for treating myocardial fibrosis due to deficiency of heart yin.

[0019] In some embodiments of the present invention, the heart yin deficiency myocardial fibrosis is accompanied by chronic heart failure.

[0020] In some embodiments of the present invention, the drug is used to improve cardiac function, inhibit cardiomyocyte hypertrophy, reduce myocardial collagen deposition, and / or downregulate the expression of fibrosis-related genes in cardiac tissue.

[0021] The present invention also discloses the application of the aforementioned traditional Chinese medicine composition in the preparation of a drug for improving symptoms of heart yin deficiency.

[0022] In some embodiments of the present invention, the symptoms of heart yin deficiency include one or more of palpitations, insomnia, five-center heat (heat in the palms, soles, and chest), and dry mouth and throat.

[0023] This invention relates to a traditional Chinese medicine composition based on the pathogenesis of "deficiency of heart yin, internal disturbance of deficiency heat, and malnourishment of the heart spirit" in Traditional Chinese Medicine, treating it with the principles of "tonifying qi and nourishing yin, clearing heat and calming the mind." The composition consists of six herbs: lily bulb, ophiopogon japonicus, ginseng, scrophularia ningpoensis, arborvitae seed, and schisandra chinensis. Lily bulb and ophiopogon japonicus are the principal herbs, nourishing the yin of the heart and lungs, moistening the meridians, and calming the mind. Ginseng is the assistant herb, tonifying qi and generating fluids, assisting the transformation and transportation of the middle jiao, and allowing yin fluids to ascend. Scrophularia ningpoensis nourishes the kidneys and reduces fire, while arborvitae seed nourishes the heart and calms palpitations; together they act as adjuvant herbs, guiding fire back to its source and calming the mind. Schisandra chinensis is the guiding herb, astringing yang with its sour properties, thus harmonizing yin and yang. The entire formula addresses the upper, middle, and lower jiao, achieving the combined effects of tonifying qi and nourishing yin, clearing heat and calming the mind.

[0024] Based on the technical solution of the present invention, the present invention has the following beneficial effects compared with the prior art:

[0025] The traditional Chinese medicine composition (Baimai Ningxin Formula) for treating myocardial fibrosis due to heart yin deficiency provided by this invention can achieve comprehensive intervention based on the principles of "tonifying qi and nourishing yin, clearing heat and calming the mind," thus addressing both the disease and its symptoms. While improving the symptoms of heart yin deficiency, it effectively inhibits the progression of myocardial fibrosis and adverse ventricular remodeling. Preliminary animal experiments and clinical observations have shown that Baimai Ningxin Formula can significantly improve cardiac function, reduce myocardial pathological damage and fibrosis, and inhibit the activation of pro-fibrotic signals at the molecular level, providing a new treatment strategy for the prevention and treatment of chronic heart failure and cardiac fibrosis related to heart yin deficiency.

[0026] Experimental studies have confirmed that the traditional Chinese medicine composition provided in this invention can significantly improve cardiac systolic function in mice with chronic heart failure, increase left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), and inhibit ventricular dilation in animal models. Furthermore, by inhibiting cardiomyocyte hypertrophy, reducing collagen deposition, and downregulating the expression of fibrosis-related genes (such as Acta2, Col1a1, Col3a1, and TGF-β), it systematically exerts protective effects against myocardial fibrosis and ventricular remodeling at the functional, tissue, and molecular levels, achieving a dual improvement in cardiac structure and function. Clinical cases show that this formula can effectively alleviate symptoms of heart yin deficiency such as palpitations, insomnia, and irritability, supporting its clinical application value in the dual intervention of "syndrome-structure."

[0027] The traditional Chinese medicine composition provided by this invention is concise and clearly defined, consisting of only six medicinal materials. The raw materials are sourced from standardized and of controllable quality, possessing a sound safety foundation and formulation feasibility. Its mechanism of action encompasses multiple levels, including functional, tissue, and genetic aspects, reflecting the multi-target and holistic regulatory characteristics of traditional Chinese medicine compound formulas. This provides experimental evidence and translational prospects for developing cardiovascular traditional Chinese medicines that combine symptom improvement and structural protection. Attached Figure Description

[0028] Figure 1 Echocardiography of cardiac function in each group of mice; among them, Figure 1 In the figure, A is a statistical bar chart of left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) for each group of mice; Figure 1 B in the figure represents a statistical bar chart of the left ventricular end-diastolic diameter (LVIDd) and end-systolic diameter (LVIDs) of each group of mice.

[0029] Figure 2 Images showing the cross-sectional area of ​​various mouse cardiomyocytes; among them, Figure 2 A in the image represents the wheat germ agglutinin (WGA) fluorescence staining image of myocardial tissue from each group of mice. Figure 2 B in the figure is a statistical bar chart of the cross-sectional area of ​​myocardial cells in each group of mice.

[0030] Figure 3 Images showing Masson staining and fibrosis detection in myocardial tissue of mice in each group; among them, Figure 3 In the diagram, A represents the Masson trichrome staining image of myocardial tissue from each group of mice. Figure 3 B in the figure is a statistical bar chart of the myocardial collagen volume fraction (CVF) of each group of mice.

[0031] Figure 4 Images of Sirius red staining for fibrosis in myocardial tissue of mice in each group; among them... Figure 4 In the diagram, A represents Sirius red staining of myocardial tissue from each group of mice. Figure 4 B in the figure represents a statistical bar chart of the myocardial fibrosis area in each group of mice.

[0032] Figure 5 The graph shows the expression of mRNA related to fibrosis in the heart tissue of mice in each group. Specifically, it is a statistical bar chart showing the relative expression levels of Acta2, Col1a1, Col3a1, and Tgfb1 genes in the heart tissue of mice in each group. Detailed Implementation

[0033] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the embodiments thereof. 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 protection scope of the present invention.

[0034] The following description is based on specific embodiments.

[0035] Experimental materials: The raw medicinal materials used in this application (lily bulb, ophiopogon japonicus, ginseng, scrophularia ningpoensis, cypress seed, and schisandra chinensis) were all purchased from the pharmacy of the First Affiliated Hospital of Guangzhou University of Chinese Medicine and were identified as meeting the standards of the Chinese Pharmacopoeia.

[0036] Example 1: A traditional Chinese medicine composition for treating myocardial fibrosis due to heart yin deficiency and its preparation method

[0037] A traditional Chinese medicine composition for treating myocardial fibrosis due to deficiency of heart yin includes 9g of lily bulb, 9g of ophiopogon japonicus, 6g of ginseng, 9g of scrophularia ningpoensis, 6g of arborvitae seed, and 3g of schisandra chinensis; this traditional Chinese medicine composition is named "Bai Mai Ning Xin Fang".

[0038] A method for preparing a traditional Chinese medicine composition for treating myocardial fibrosis due to deficiency of heart yin, using a decoction method:

[0039] The following herbs were weighed according to the prescription (Bai Mai Ning Xin Fang): 9g lily bulb, 9g ophiopogon japonicus, 6g ginseng, 9g scrophularia ningpoensis, 6g arborvitae seed, and 3g schisandra chinensis. After mixing, the herbs were soaked in 10 times their volume of distilled water for 30 minutes, brought to a boil over high heat, then simmered over low heat for 40 minutes. The mixture was filtered, and the filtrate was collected. The residue was then soaked in 8 times its volume of distilled water for a second time, brought to a boil over high heat, then simmered over low heat for 30 minutes. The filtrates from both processes were combined. The combined filtrate was concentrated using a rotary evaporator to obtain different concentrations of Bai Mai Ning Xin Fang decoction. Experimental animals were administered the decoction via gavage, with each mouse receiving 0.2 mL per dose. Three dosage levels (low, medium, and high) were established based on the decoction concentration, corresponding to crude drug doses of 2.73, 5.46, and 10.92 g / kg·d, respectively. The Bai Mai Ning Xin Fang decoction was stored at 4℃ for later use. Before administration, the decoction was warmed to room temperature and mixed thoroughly.

[0040] Example 2: A traditional Chinese medicine composition for treating myocardial fibrosis due to heart yin deficiency and its preparation method

[0041] A traditional Chinese medicine composition for treating myocardial fibrosis includes 5g of lily bulb, 5g of ophiopogon japonicus, 3g of ginseng, 5g of scrophularia ningpoensis, 3g of arborvitae seed, and 1g of schisandra chinensis; this traditional Chinese medicine composition is named "Bai Mai Ning Xin Fang".

[0042] The preparation method is the same as in Example 1.

[0043] Example 3: A traditional Chinese medicine composition for treating myocardial fibrosis due to heart yin deficiency and its preparation method

[0044] A traditional Chinese medicine composition for treating myocardial fibrosis includes 13g of lily bulb, 13g of ophiopogon japonicus, 9g of ginseng, 13g of scrophularia ningpoensis, 9g of arborvitae seed, and 5g of schisandra chinensis; this traditional Chinese medicine composition is named "Bai Mai Ning Xin Fang".

[0045] The preparation method is the same as in Example 1.

[0046] Experimental Example: Pharmacodynamic Study of the Drug Solution for Treating Myocardial Fibrosis Due to Heart Yin Deficiency

[0047] The traditional Chinese medicine decoction was prepared according to the method shown in Example 1 for efficacy studies. Specifically, in order to study the therapeutic effect of the traditional Chinese medicine composition provided in this application on myocardial fibrosis due to heart yin deficiency, a mouse model of chronic heart failure (CHF) was established, and the mouse was treated with the Bai Mai Ning Xin Fang formula provided in this application. Then, efficacy evaluation and pharmacological analysis were performed.

[0048] 1. Laboratory animals

[0049] A total of 30 SPF-grade male C57BL / 6J mice (8 weeks old, weighing 16–20 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. All mice were housed under conditions of temperature (22–25 °C), humidity (40–60%), a 12-h light / dark cycle, with free access to food and water. After ≥7 d of adaptive feeding, the mice were used in the experiment (license number: SCXK (Guangdong) 2022-0002). The mice were randomly divided into 6 groups (n = 5 per group) using a random number table method / computer random method: control group, model group, low-dose BMNX group (BMNX-L), medium-dose BMNX group (BMNX-M), high-dose BMNX group (BMNX-H), and ARNI group. During the experiment, data collection and analysis were performed using a blinded method.

[0050] 2. Animal model establishment

[0051] A chronic heart failure (CHF) model was established using ligation of the left anterior descending coronary artery (LAD). After inhalation anesthesia with isoflurane, the mice were fixed on the operating table, tracheally intubated, and connected to a small animal ventilator for assisted breathing. The chest was opened between the 3rd and 4th left intercostal spaces to expose the heart. At the lower edge of the left atrial appendage along the coronary artery, the LAD was ligated with 8-0 prolene suture. Successful ligation was indicated by the whitening of the anterior wall of the left ventricle and weakened movement. Subsequently, the chest cavity and skin were sutured layer by layer. After the operation, the mice were kept warm and resuscitated, and perioperative treatments such as analgesia and anti-infection were carried out. The control group underwent sham surgery (only thoracotomy and threading were performed, without ligating the LAD, and the remaining steps were the same as those in the model group).

[0052] 3. Grouping and drug administration

[0053] After successful model establishment, the 30 mice were randomly divided into 6 groups, with 5 mice in each group. The specific grouping and drug administration regimens are shown in Table 1:

[0054] Table 1 Animal grouping and drug administration

[0055]

[0056] After the mice recovered stably after surgery, drug administration began. The drugs were administered by gavage once a day for 4 consecutive weeks.

[0057] 4. Experimental methods

[0058] 4.1 General condition observation

[0059] The mental state, activity level, hair luster, nasal and oral color, auricle vascular filling degree, paw pad / tail color, water intake, and fecal characteristics of the mice were observed and recorded daily.

[0060] 4.2 Echocardiography for cardiac function detection

[0061] After the intervention, small animal echocardiography was performed to assess cardiac function. Mice were anesthetized to a light state (maintaining a relatively stable heart rate and avoiding underestimation of cardiac function due to excessive anesthesia), their chest hair was removed, and they were placed on a temperature-controlled platform connected to an electrocardiogram monitor. Two-dimensional (B-mode) and M-mode images were acquired at the level of the left ventricular papillary muscles, and ≥3 consecutive stable cardiac cycles were obtained and averaged. Parameters measured and calculated included: left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), end-diastolic left ventricular diameter (LVIDd), and end-systolic left ventricular diameter (LVIDs).

[0062] 4.3 Myocardial tissue pathological examination

[0063] After ultrasound examination, mice were euthanized by overdose and their hearts were perfused. The hearts were then rapidly removed. The hearts were fixed in 4% paraformaldehyde for 24 hours, followed by routine dehydration, clearing, and paraffin embedding. Serial sections were prepared along the short axis of the heart for the following staining:

[0064] (1) WGA fluorescence staining: to assess the cross-sectional area of ​​cardiomyocytes. After dewaxing and rehydration of paraffin sections, FITC-labeled wheat germ agglutinin (WGA, 10 μg / mL) working solution was added, and the sections were incubated at room temperature in the dark for 1 hour. The cell nuclei were counterstained with DAPI, and the sections were mounted and observed and photographed under a fluorescence microscope. The cross-sectional area of ​​at least 30 cardiomyocytes was randomly measured using ImageJ software, and the average value was used for statistical analysis.

[0065] (2) Masson's trichrome staining: to assess the degree of myocardial fibrosis. Follow the instructions of the kit (Pinofi Biotechnology Co., Ltd., P0043). Collagen fibers appear blue, and myocardial fibers appear red. Take photos under a microscope and calculate the collagen volume fraction (CVF) = collagen area / total myocardial area × 100%.

[0066] (3) Sirius red staining: to assess the area of ​​collagen deposition and fibrosis. Following the instructions of the kit (Pinofi Biotechnology Co., Ltd., S191018), collagen fibers appeared red. Images were taken under a microscope under polarized light, and the percentage of fibrosis area was calculated.

[0067] 4.4 Gene expression detection (qPCR)

[0068] Left ventricular tissue from mice was flash-frozen in liquid nitrogen and stored at -80 °C. Total RNA was extracted, and RNA concentration and purity (A260 / 280) were measured. cDNA was synthesized using a reverse transcription kit (EZB, Cat. No. EZB-RN001-plus). Real-time quantitative PCR was used to detect the mRNA expression of fibrosis-related genes, including Acta2, Col1a1, Col3a1, and Tgfb1. Gapdh was used as an internal control gene, and a 2...-ΔΔCt The relative expression levels of the target gene were calculated using primer sequences shown in Table 2. Each sample was configured with technical replicates and included a template-free control to exclude contamination.

[0069] Table 2 Primer sequence list

[0070]

[0071] 4.5 Data Analysis Methods

[0072] Normally distributed data are expressed as mean ± standard deviation (x ± s), and non-normally distributed data are expressed as median (interquartile range, M [P25–P75]). All statistical analyses were performed using GraphPad Prism software (GraphPad Software, San Diego, CA, USA). Normality of the data was assessed using the Shapiro–Wilk test, and homogeneity of variance was tested. For non-normally distributed data, the Mann–Whitney U test was used for comparisons between two groups, and the Kruskal–Wallis test was used for comparisons among multiple groups, with Dunn's post-hoc test performed when necessary. For normally distributed data, Student's t test or Welch's t test was used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. When homogeneity of variance was satisfied, the Tukey HSD method was used for post-hoc comparisons; when variances were unequal, Dunnett's T3 method was used for comparisons between groups. A p-value < 0.05 is considered statistically significant.

[0073] 5 Experimental Results

[0074] 5.1 The traditional Chinese medicine composition of this application improves the general condition of mice with heart failure.

[0075] Compared with the control group, the general condition of CHF model mice gradually declined after surgery, exhibiting manifestations consistent with "yin deficiency and internal heat": lethargy, reduced activity, and sluggish response; decreased coat luster, dryness and yellowing (some with piloerection); dark red mouth and nose, obvious engorgement of blood vessels in the auricles, and darkening and loss of luster in the paw pads and tail. Some mice showed increased licking around the mouth, more frequent drinking, and drier feces. The auricles / paw pads were warmer to the touch or showed a "preferring coolness" tendency, suggesting external characteristics of excessive internal heat and insufficient body fluids.

[0076] After intervention with Bai Mai Ning Xin Fang, the general condition of mice in each dosage group improved to varying degrees compared with the model group, showing improved mental state and activity, enhanced responsiveness; reduced dry and dull fur, alleviated dark red mouth and nose and vascular engorgement of auricles, and improved luster of paw pads / tails; some mice also showed an improvement trend in frequent water drinking and dry feces, suggesting that Bai Mai Ning Xin Fang helps to correct the body fluid deficiency-like manifestations after "yin deficiency and internal heat".

[0077] 5.2 The traditional Chinese medicine composition of this application improves cardiac dysfunction in mice with heart failure.

[0078] After modeling, small animal echocardiography was used to assess the cardiac function of mice in each group. For example... Figure 1 The echocardiogram results of A and B showed that, compared with the control group, the model group mice had significant impaired cardiac systolic function, manifested by a significant decrease in left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (P < 0.0001), while the left ventricular diameter showed dilatational changes, with significant increases in both end-diastolic left ventricular diameter (LVIDd) and end-systolic left ventricular diameter (LVIDs) (P < 0.0001), indicating that the CHF mouse model was successfully established.

[0079] After intervention with Baimaining Xinfang, the cardiac function indicators of mice showed varying degrees of improvement compared with the model group: LVEF increased compared with the model group (P<0.0001), LVFS increased compared with the model group (P<0.001), accompanied by a decrease in LVIDs (P<0.001) and LVIDd (P<0.05), suggesting that Baimaining Xinfang can effectively alleviate the impaired ventricular systolic function and ventricular dilatational remodeling in CHF model mice.

[0080] The above results indicate that Bai Mai Ning Xin Fang can improve left ventricular systolic function and protect cardiac function in CHF model mice.

[0081] 5.3 The traditional Chinese medicine composition of this application inhibits cardiomyocyte hypertrophy in mice with heart failure.

[0082] To further evaluate cardiomyocyte hypertrophy in CHF model mice, wheat germ agglutinin (WGA) staining was used to observe the cross-sectional morphology of cardiomyocytes and to perform quantitative analysis of cross-sectional area. Figure 2 The results of the cross-sectional area detection of cardiomyocytes in mice A and B showed that the cross-sectional area of ​​cardiomyocytes in the model group was significantly larger than that in the control group (P < 0.0001), suggesting that cardiac hypertrophy occurred in the CHF model mice.

[0083] After intervention with Baimaining Xinfang, the degree of cardiomyocyte hypertrophy was significantly reduced compared with the model group, as evidenced by a significant decrease in the cross-sectional area of ​​cardiomyocytes (P<0.05) and a more regular myocardial structure compared with the model group, suggesting that it can effectively inhibit the process of myocardial hypertrophy in CHF model mice.

[0084] The results suggest that, in addition to improving cardiac function, Baimainingxinfang can also inhibit cardiomyocyte hypertrophy in CHF model mice, thereby delaying adverse ventricular remodeling.

[0085] 5.4 The traditional Chinese medicine composition of this application improves myocardial fibrosis and collagen deposition in mice with heart failure.

[0086] To assess the degree of myocardial fibrosis in CHF model mice, Masson staining and Sirius red staining were used to observe and quantify the levels of myocardial collagen deposition and fibrosis. Figure 3 Masson staining of myocardial tissue from mice A and B showed a significant increase in interstitial collagen fibers in the model group (P < 0.0001), with an expanded area of ​​blue-stained collagen deposition, indicating significant excessive collagen deposition and interstitial fibrosis in the CHF model mice. After intervention with medium and high doses of Baimaining Xinfang, myocardial collagen deposition was significantly reduced compared to the model group (P < 0.0001), and the fibrotic area shrank, suggesting that it can effectively alleviate abnormal collagen accumulation in the myocardium of CHF model mice.

[0087] Furthermore, such as Figure 4 The results of A and B Sirius red staining also showed that the degree of myocardial fibrosis in CHF model mice was significantly increased (P < 0.0001), manifested as enhanced collagen fiber deposition and expanded fibrosis area; while the intervention of Baimaining Xinfang could significantly reduce Sirius red positive collagen deposition and significantly alleviate fibrosis level (P < 0.001), suggesting that it has a clear ameliorative effect on myocardial fibrosis in CHF model mice.

[0088] In conclusion, Baimainingxinfang can reduce collagen deposition and fibrosis in the heart of CHF model mice, suggesting that it has an anti-fibrotic protective effect against CHF-related structural remodeling.

[0089] 5.5 The herbal composition of this application inhibits the expression of profibrosis-related genes in mice with heart failure.

[0090] The mRNA expression levels of fibrosis-related genes Acta2, Col1a1, Col3a1, and TGF-β in cardiac tissue were detected using qPCR. Figure 5 qPCR results showed that, compared with the control group, the expression of the above-mentioned fibrosis-related genes in CHF model mice was significantly upregulated (P < 0.0001), suggesting that CHF model mice have significantly enhanced transcriptional activation and collagen production-related programs in cardiac fibrosis.

[0091] After intervention with Baimaining Xinfang, the expression of fibrosis-related genes in each dose group decreased to varying degrees compared with the model group, showing an overall trend of increasing inhibitory effect with increasing dose. Among them, the BMNX-H group showed the most prominent inhibitory effect, significantly downregulating the mRNA expression of Acta2, Col1a1, Col3a1, and TGF-β (P < 0.001), suggesting that high-dose Baimaining Xinfang can more effectively inhibit the expression of cardiac fibrosis-related genes in CHF model mice.

[0092] The above results indicate that the herbal composition of this application can inhibit the abnormal upregulation of myocardial fibrosis-related genes in CHF mice, which is consistent with the histological results of improving collagen deposition and reducing fibrosis. This suggests that it may reduce adverse ventricular remodeling by inhibiting pro-fibrosis signals and collagen production processes.

[0093] Clinical application cases

[0094] Case 1

[0095] Patient Huang, female, 62 years old, had been experiencing palpitations for over a year, characterized by episodic episodes lasting one minute each. She was diagnosed at another hospital with "occasional premature ventricular contractions and anxiety," and was regularly taking bisoprolol and duloxetine. Later, due to insomnia, she took eszopiclone and estazolam to aid sleep. After being given a traditional Chinese medicine compound formula to nourish Yin and calm the mind, her palpitations improved slightly. At the time of consultation, she presented with occasional dizziness, fatigue, good appetite, hot palms, loose stools, frequent urination at night, and a red, dry tongue with a thin white coating. These symptoms met the TCM diagnostic criteria for palpitations and insomnia, and the syndrome was identified as Yin deficiency of the heart. She was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250mL, taken in two divided doses morning and evening, for 14 consecutive days. After treatment, the patient's palpitations improved, the frequency of attacks decreased, dizziness slightly improved, sleep duration increased, and the hot palm symptoms improved.

[0096] Case 2

[0097] Patient Deng, female, 74 years old, presented with palpitations for over a month, accompanied by chest tightness, which was more pronounced during sleep at night, recurring and lasting for one hour each time, accompanied by dizziness and headache. She had a history of hypertension and diabetes. Upon consultation, her symptoms included palpitations, chest tightness, normal appetite, poor sleep, easy awakening, palpitations upon waking, normal urination, and dry stools. This met the diagnostic criteria for palpitations and insomnia in Traditional Chinese Medicine (TCM), and was identified as a syndrome of heart yin deficiency. She was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250mL, taken in two divided doses morning and evening, for five consecutive days. After treatment, the patient's palpitations significantly improved, and her sleep slightly improved. After continuing treatment for another seven days, her sleep disturbances also improved.

[0098] Case 3

[0099] Patient Li, male, 42 years old, had previously undergone catheter radiofrequency ablation for Wolff-Parkinson-White syndrome with palpitations. He relapsed half a month post-procedure, experiencing a heart rate of 190 bpm during each episode, lasting approximately half an hour. He reported palpitations, shortness of breath, fatigue, chest pain, and dry mouth. His watch monitoring revealed premature ventricular contractions (PVCs). He also experienced decreased exercise tolerance, poor sleep, and difficulty falling asleep. His diagnosis of palpitations conformed to Traditional Chinese Medicine (TCM) principles, specifically a pattern of Yin deficiency in the heart. He was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250 mL, taken in two divided doses morning and evening, for 14 consecutive days. After treatment, the patient's palpitations improved, the frequency of attacks decreased, and a heart rate exceeding 120 bpm was no longer detected. PVCs significantly decreased, sleep improved, and shortness of breath and fatigue subsided.

[0100] Case 4

[0101] Patient Guo, female, 73 years old, had a history of recurrent heartburn for over two years, accompanied by acid reflux, belching, and recurrent palpitations. A gastroscopy at a local hospital revealed gastroesophageal reflux and multiple proliferative gastric polyps. Coronary angiography showed 55% stenosis of the left anterior descending artery. She had no significant chest tightness, chest pain, or decreased exercise tolerance. Upon presentation, her symptoms included acid reflux, heartburn, bloating, belching, irritability, palpitations, dry mouth, bitter taste, fair appetite, fair sleep, and normal bowel movements. This met the diagnostic criteria of Traditional Chinese Medicine (TCM) for acid reflux and palpitations, and was diagnosed as a syndrome of heart yin deficiency. She was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250mL, taken in two divided doses morning and evening, for 14 consecutive days. After treatment, the patient's acid reflux symptoms improved, the frequency decreased, bloating and belching lessened, appetite increased, and palpitations did not recur.

[0102] Case 5

[0103] Patient Kuang, male, 28 years old, had been experiencing chest tightness and palpitations for over half a month. He was initially diagnosed with "acute myocarditis" due to "chest tightness, palpitations, and recurrent low-grade fever," and had been taking trimetazidine and metoprolol. Upon consultation, he still experienced chest tightness and palpitations, restlessness, shortness of breath, poor sleep with vivid dreams, dry mouth, bitter taste, normal appetite, and normal bowel movements. This met the diagnostic criteria for palpitations in Traditional Chinese Medicine (TCM), and the syndrome was identified as Yin deficiency of the heart. He was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250mL, taken in two divided doses morning and evening, for 14 consecutive days. After treatment, the patient's chest tightness and palpitations improved, and his restlessness significantly decreased. He still had poor sleep, which improved after another 7 doses.

[0104] Case 6

[0105] Patient Lin, female, 52 years old, presented with chest tightness and pain for over three months. She had suffered a left rib fracture due to a sprain, resulting in persistent left chest pain accompanied by chest tightness. At the time of consultation, she still experienced chest tightness and pain, which came and went intermittently. She also experienced poor sleep, difficulty falling asleep, frequent awakenings, dry mouth, belching, irritability, and frequent sighing. Her condition met the diagnostic criteria for hypochondriac pain in Traditional Chinese Medicine (TCM), and the syndrome was identified as Yin deficiency of the heart. She was prescribed the herbal decoction prepared in Example 1, one dose daily, decocted to 250mL, taken in two divided doses morning and evening, for 14 consecutive days. After treatment, the patient's mood improved, the chest tightness and pain lessened, and her sleep improved slightly.

[0106] Case 7

[0107] Patient Chen, male, 62 years old, had repeated chest tightness and palpitations for more than 5 years, with the symptoms aggravated for 1 week. He was diagnosed with coronary heart disease in an external hospital. When seeking medical treatment, he felt a sense of stuffiness in the manubrium sterni, especially at night, and numbness in both palms from time to time. His appetite was normal, sleep was average, his lips were dark, his tongue was dark red, and the sublingual collaterals were tortuous. It conformed to the diagnosis of chest impediment and heart pain in traditional Chinese medicine, and was differentiated as the syndrome of heart yin deficiency. He was given the traditional Chinese medicine decoction prepared in Example 1, 1 dose per day, decocted to 250 mL, taken in the morning and evening, and treated continuously for 14 days. After treatment, the patient's symptoms of chest tightness and palpitations improved, the numbness symptoms in both palms were slightly improved, and the sleep was improved compared with before.

[0108] To sum up, as a traditional Chinese medicine compound composition with relatively clear composition and convenient quality control, the traditional Chinese medicine composition (Baimai Ningxin Formula) provided by the present invention has the following beneficial technical effects in the treatment of myocardial fibrosis related to heart yin deficiency, and can be explained from the technical principle level of "combining disease and syndrome":

[0109] First of all, from the perspective of traditional Chinese medicine syndromes, Baimai Ningxin Formula is based on the therapeutic principles of "tonifying qi and nourishing yin, nourishing yin and clearing heat, and calming the mind and tranquilizing the nerves", and can play a comprehensive regulatory role in the core pathogenesis of "heart yin deficiency, internal disturbance of deficiency heat, and loss of nourishment of the mind" of the syndrome of heart yin deficiency, which helps to improve the clinical manifestations related to heart yin deficiency, such as palpitations, sleep disorders, restlessness, internal heat, and dry mouth. The above-mentioned typical case shows that after giving the compound decoction of the present invention, the frequency of palpitations in the patient decreased, the sleep time was prolonged or the improvement of waking up easily was achieved, and the restlessness and fever in the palms were reduced, suggesting that the present invention can improve the core symptoms of heart yin deficiency to a certain extent and meet the treatment needs at the "symptom - syndrome" level in the long-term management of chronic diseases.

[0110] Secondly, from the perspective of modern medical pathology and phenotype, in LAD-ligated CHF model mice, intervention with Bai Mai Ning Xin Fang not only improved the general condition changes consistent with "Yin deficiency and internal heat, fluid depletion" in the model animals, but also showed a protective effect against cardiac dysfunction and structural remodeling defects. Specifically, in terms of general condition, common physical and behavioral changes in model mice, such as decreased mental activity, dry and dull fur, dark red mouth and nose, engorged ear blood vessels, darkened paw pads / tail color, as well as frequent drinking, dry feces, and increased temperature sensitivity / preferring coolness, were alleviated to varying degrees after intervention with Bai Mai Ning Xin Fang. Regarding cardiac function and structure, echocardiography showed that it could improve left ventricular systolic function and alleviate dilatational remodeling (increased LVEF and LVFS, decreased LVIDd / LVIDs); histological results showed that it could reduce cardiomyocyte hypertrophy (decreased WGA cross-sectional area) and reduce collagen deposition and fibrosis levels (reduced Masson and Sirius red positive areas); at the molecular level, qPCR further showed that it could downregulate the expression of pro-fibrosis-related genes (Acta2, Col1a1, Col3a1, Tgfb1). The above functional, histological, and molecular evidence are consistent, suggesting that Baimaining Xinfang can, to a certain extent, inhibit myocardial fibrosis and adverse remodeling during the progression of CHF, thereby achieving comprehensive protection of cardiac function and structure.

[0111] In terms of technical principles, the beneficial effects of this invention stem from its compound synergistic and holistic regulatory characteristics: on the one hand, by nourishing the heart's yin, clearing away deficiency heat, and calming the mind, it helps correct the persistent imbalance corresponding to "yin deficiency and internal heat—malnourishment of the collaterals," thus improving the underlying syndrome of heart yin deficiency; on the other hand, by inhibiting key processes related to fibrosis during the progression of heart failure (such as myofibroblast activation and collagen generation and deposition), it reduces myocardial interstitial fibrosis and delays adverse ventricular remodeling, ultimately resulting in improved cardiac function and structural protection. Therefore, this invention achieves an integrated approach to both "improvement of heart yin deficiency syndrome" and "intervention in the risk of cardiac fibrosis / remodeling," overcoming the shortcomings of existing technologies that only focus on symptom control or struggle to address structural remodeling intervention.

[0112] Therefore, this invention provides experimental evidence and theoretical support for the treatment of myocardial fibrosis related to Yin deficiency of the heart, and lays the foundation for subsequent clinical translation and application.

[0113] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0114] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A traditional Chinese medicine composition for treating myocardial fibrosis due to deficiency of heart yin, characterized in that, The traditional Chinese medicine composition consists of the following raw medicinal materials in parts by weight: 5-13 parts lily bulb, 5-13 parts ophiopogon japonicus, 3-9 parts ginseng, 5-13 parts scrophularia ningpoensis, 3-9 parts arborvitae seed, and 1-5 parts schisandra chinensis.

2. The traditional Chinese medicine composition as described in claim 1, characterized in that, It is composed of the following raw medicinal materials in parts by weight: 7-11 parts lily bulb, 7-11 parts ophiopogon japonicus, 5-7 parts ginseng, 7-11 parts scrophularia ningpoensis, 5-7 parts arborvitae seed, and 2-4 parts schisandra chinensis.

3. The method for preparing the traditional Chinese medicine composition as described in claim 1 or 2, characterized in that, include: Take the raw medicinal materials of the Chinese herbal composition according to the formula, extract with water, and collect the extract to obtain the final product.

4. A drug for treating myocardial fibrosis due to deficiency of heart yin, characterized in that, The drug uses the traditional Chinese medicine composition as described in claim 1 or 2 as its active ingredient.

5. The drug as described in claim 4, characterized in that, The medicine is in the form of decoction, granules, pills, capsules, tablets, powders, or oral liquid.

6. The drug as described in claim 4, characterized in that, The drug also includes other pharmaceutically acceptable excipients.

7. The drug as described in claim 4, characterized in that, The drug also includes a drug carrier.

8. The use of the traditional Chinese medicine composition as described in claim 1 or 2 in the preparation of a medicament for treating myocardial fibrosis due to deficiency of heart yin.

9. The application as described in claim 8, characterized in that, The drug is used to improve cardiac function, inhibit cardiomyocyte hypertrophy, reduce myocardial collagen deposition, and / or downregulate the expression of fibrosis-related genes in cardiac tissue.

10. The use of the traditional Chinese medicine composition as described in claim 1 or 2 in the preparation of a medicament for improving symptoms of heart yin deficiency.

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