A traditional Chinese medicine formula for treating cardiac impairment and heart failure caused by anthracycline chemotherapy drugs, and a preparation method and application thereof

Abstract

The application discloses a traditional Chinese medicine formula for treating heart damage and heart failure caused by anthracycline chemotherapy drugs and a preparation method and application thereof. The traditional Chinese medicine formula comprises the following components in parts by mass: 50-70 parts of raw astragalus, 10-30 parts of rhodiola, 3-8 parts of prepared aconite root, 5-15 parts of polyporus umbellatus, 5-15 parts of Semen Lepidii, 5-15 parts of curcuma zedoary, 5-18 parts of white peony root and 1-7 parts of dried ginger. The traditional Chinese medicine formula can improve heart damage and heart failure caused by anthracycline chemotherapy drugs, significantly inhibit the interventricular septum thickness during diastole and systole, left ventricular posterior wall thickness during diastole and systole of DOX-HF mice, improve the left ventricular ejection fraction of the mice, effectively relieve the change of myocardial cell morphology and cardiac fibrosis caused by DOX, correct the decrease of acylcarnitine content caused by DOX, reverse the inhibition of DOX on the fatty acid beta oxidation function of heart tissue, and thus reduce the heart damage caused by anthracycline chemotherapy drugs.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine formula, preparation method, and application for treating cardiac damage and heart failure caused by anthracycline chemotherapy drugs, belonging to the field of traditional Chinese medicine. Background Technology

[0002] Anthracyclines (doxorubicin, daunorubicin, epirubicin, and idarubicin) are widely used as first-line treatment for various cancers, including breast cancer, sarcoma, lymphoma, and leukemia. Although anthracyclines are highly effective and difficult to replace, their dose-dependent cardiotoxicity can significantly increase the incidence of heart failure (HF) in cancer patients. HF induced by anthracycline chemotherapy drugs is currently a hot topic in cardiac oncology research. Extensive epidemiological and evidence-based medicine evidence has shown that even with standard doses of doxorubicin (DOX) for breast cancer treatment, approximately 20% of patients experience decreased left ventricular systolic function (left ventricular ejection fraction decrease >10%) within the first 6 months of treatment, and this decrease increases when the cumulative treatment dose rises to 700 mg / m². 2 At that time, the incidence of HF can reach nearly 50%.

[0003] Dexrazoxane is a drug that can clearly improve DOX-HF and is currently the only drug approved by the US FDA for the prevention of anthracycline cardiotoxicity. Its main mechanism of action is to compete with DOX for Top2β. Related reports suggest that it may reduce the chemotherapeutic effect of anthracyclines and has the potential risk of causing secondary malignancies, which limits the clinical application of dexrazoxane. Besides dexrazoxane, various cardiovascular disease prevention and treatment drugs, especially sodium-glucose cotransporter-2 inhibitors, beta-blockers, angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, spironolactone, and statins, have shown preliminary evidence of potential preventive effects against DOX-HF, but their efficacy is still unclear, and clinical studies are insufficient, requiring further evidence. To protect the active drug from systemic circulation and increase the specificity of tumor cell uptake, a pegylated liposome form of doxorubicin has been developed, and its clinical efficacy has been demonstrated, but it has not yet shown a significant reduction in the incidence of DOX-HF. Furthermore, the high cost is also a limiting factor for liposomal administration; many clinicians also use epirubicin instead of doxorubicin in hopes of reducing cardiotoxicity, but studies have shown that there is no significant difference between the two drugs when used at equivalent doses. Clearly, there is currently a lack of economically available and effective Chinese and Western medicines for the treatment of anthracycline-induced cardiac damage and heart failure. Therefore, exploring the potential mechanisms of DOX-HF and seeking safe and effective treatments is of significant clinical and economic importance for improving the prognosis of patients using anthracycline chemotherapy drugs and reducing the incidence of HF. Summary of the Invention

[0004] Purpose of the invention: The technical problem to be solved by the present invention is to provide a traditional Chinese medicine formula, preparation method and application for treating cardiac damage and heart failure caused by anthracycline chemotherapy drugs.

[0005] Technical solution: To solve the above-mentioned technical problems, the present invention provides a traditional Chinese medicine composition for invigorating qi and warming yang, comprising the following components by mass: 50-70 parts of raw astragalus, 10-30 parts of rhodiola rosea, 3-8 parts of prepared aconite root, 5-15 parts of polyporus umbellatus, 5-15 parts of lepidium seed, 5-15 parts of turmeric, 5-18 parts of white peony root, and 1-7 parts of dried ginger.

[0006] The formula, by weight, includes the following components: 55-65 parts of raw Astragalus membranaceus, 15-25 parts of Rhodiola rosea, 3-6 parts of processed Aconitum carmichaelii, 10-15 parts of Polyporus umbellatus, 10-15 parts of Lepidium apetalum, 10-15 parts of Curcuma zedoaria, 10-18 parts of Paeonia lactiflora, and 1-5 parts of dried ginger.

[0007] The formula, by weight, includes the following components: 60 parts raw Astragalus membranaceus, 20 parts Rhodiola rosea, 6 parts prepared Aconitum carmichaelii, 10 parts Polyporus umbellatus, 10 parts Lepidium apetalum, 10 parts Curcuma zedoaria, 12 parts Paeonia lactiflora, and 3 parts dried ginger.

[0008] The present invention also provides a traditional Chinese medicine preparation comprising the aforementioned traditional Chinese medicine composition.

[0009] This also includes pharmaceutically acceptable excipients.

[0010] The present invention also provides a freeze-dried powder containing the aforementioned traditional Chinese medicine composition that invigorates qi and warms yang.

[0011] This invention also provides a method for preparing the freeze-dried powder described above. The method involves soaking the herbal composition containing the invigorating and warming properties of traditional Chinese medicine in water, bringing it to a boil over high heat, then simmering it over low heat, and collecting the first decoction. Cold water is added to the dregs, and the mixture is brought to a boil again, then simmered over low heat, and the second decoction is collected. The two decoctions are mixed and heated to concentrate the mixture to half its original volume. The mixture is then poured out and coarsely filtered through a double-layered gauze to obtain the herbal decoction. The decoction is then placed in a freeze dryer for vacuum freeze-drying to obtain the freeze-dried powder containing the invigorating and warming properties of traditional Chinese medicine.

[0012] The present invention also provides the application of the traditional Chinese medicine composition that invigorates qi and warms yang, or the traditional Chinese medicine preparation or the freeze-dried powder, in the preparation of a drug for treating pharmacological cardiac injury.

[0013] The present invention also provides the application of the traditional Chinese medicine composition that invigorates qi and warms yang, or the traditional Chinese medicine preparation or the freeze-dried powder, in the preparation of a drug for treating pharmacological heart failure.

[0014] The present invention also provides the use of the traditional Chinese medicine composition, the traditional Chinese medicine preparation, or the lyophilized powder in the preparation of a medicament for treating cardiac damage caused by anthracycline chemotherapy drugs.

[0015] The present invention also provides the use of the traditional Chinese medicine composition, the traditional Chinese medicine preparation, or the lyophilized powder in the preparation of a drug for treating heart failure caused by anthracycline chemotherapy drugs.

[0016] The present invention also provides the application of the traditional Chinese medicine composition for invigorating qi and warming yang, or the traditional Chinese medicine preparation or the lyophilized powder in the preparation of a drug for treating heart failure caused by doxorubicin.

[0017] The present invention also provides the use of the traditional Chinese medicine composition that invigorates qi and warms yang, or the traditional Chinese medicine preparation or the freeze-dried powder, in the preparation of a medicament for treating cardiac damage caused by doxorubicin.

[0018] DOX-HF can be roughly classified as heart failure in Traditional Chinese Medicine Internal Medicine. Anthracycline chemotherapy drugs can be understood as highly toxic substances in the concept of Chinese medicine. If a person with a weak constitution due to tumors uses them for a long time, it will inevitably damage the heart, resulting in a deficiency of heart yang and a lack of nourishment for the heart. Ultimately, both the body and function will be damaged, leading to heart failure. The root cause of its pathogenesis is the deficiency of qi and yang. When qi and yang (heart function) are exhausted and unable to support themselves, the circulation of qi, blood and body fluids will be impaired. "When blood is not flowing smoothly, it becomes water." Subsequently, blood stasis and dampness combine to form symptoms, ultimately leading to the root and symptoms of heart failure. Based on the core pathogenesis of Qi and Yang deficiency and the mutual binding of stagnant water, the treatment principle of tonifying Qi and warming Yang, resolving stasis and promoting diuresis is determined. The formulas such as Linggui Zhugan Decoction, Zhigancao Decoction, and Zhenwu Decoction in the "Treatise on Cold Damage" all reflect the intention of tonifying Qi and warming Yang. The "Synopsis of Prescriptions of the Golden Chamber" states that "for those with phlegm and fluid retention, warm medicines should be used to harmonize them." Later, the "Standards for Diagnosis and Treatment" mentions that "if the heart Qi is insufficient and the kidney water overwhelms it, causing it to rise and stop in the heart, it is necessary to stop the rebellious Qi, drain the water, and tonify the Yang." All of these are in line with this treatment principle. The heart Yang originates from the kidney Yang, and the kidney Yang is the root of the Yin and Yang of the whole body. Therefore, invigorating the heart Yang Qi and restoring the warming effect of the kidney Yang, warming Yang and transforming Qi to circulate throughout the body, and helping to disperse stagnant water, treating both the root cause and the symptoms, is the main treatment method for this disease.

[0019] The Qi-tonifying and Yang-warming formula consists of eight Chinese herbs: raw astragalus, rhodiola rosea, prepared aconite root, polyporus umbellatus, lepidium seed, turmeric, white peony root, and fresh ginger. Raw astragalus tonifies qi and promotes diuresis, serving as the principal herb. Rhodiola rosea tonifies qi, nourishes the heart, and invigorates blood; prepared aconite root warms and tonifies the heart and kidneys, serving as the assistant herbs. Polyporus umbellatus and lepidium seed purge the lungs and promote diuresis; turmeric breaks up blood stasis and promotes qi circulation, serving as the adjuvant herbs. White peony root and fresh ginger act as the guiding herbs. The entire formula works synergistically to tonify qi and warm yang, resolve blood stasis, and promote diuresis. Based on tonifying heart qi and warming heart yang, it further utilizes polyporus umbellatus and lepidium seed to promote diuresis, and turmeric to promote qi circulation and break up blood stasis, ensuring that tonification does not lead to stagnation. This balanced approach of tonifying and draining, with half the effort yielding twice the result.

[0020] To further explore the pharmacological effects and mechanisms of Yiqi Wenyang Formula in treating DOX-HF, this invention constructed an animal model of DOX-HF and intervened with Yiqi Wenyang Formula. It was found that Yiqi Wenyang Formula can significantly reduce LVIDs in DOX-HF mice, increase EF% and FS% in DOX-HF mice, and effectively alleviate the changes in cardiomyocyte morphology and cardiac fibrosis caused by DOX. Acylcarnitine is an intermediate product of fatty acid β-oxidation. In this invention, non-target metabolomics analysis was performed on mid-section of mouse heart tissue. The results showed that DOX significantly reduced the levels of acylcarnitine Stearoyl-L-Carnitine (Acylcarnitine C18:0), L-Palmitoylcarnitine (C16Carnitine), Tetradecanoyl-L-Carnitine (C14 Carnitine), Octanoyl-L-Carnitine (C8 Carnitine), and Hexanoyl-L-Carnitine (C6 Carnitine) in heart tissue, indicating that fatty acid β-oxidation in heart tissue was inhibited. The Qi-invigorating and Yang-tonifying formula could correct the reduction in acylcarnitine levels caused by DOX, suggesting that the Qi-invigorating and Yang-tonifying formula may reverse the inhibition of fatty acid β-oxidation function in heart tissue by DOX.

[0021] Beneficial Effects: Compared with the prior art, the present invention has the following significant advantages: 1. The Qi-invigorating and Yang-tonifying formula of the present invention has good clinical efficacy for heart failure, including improving the TCM syndrome score of heart failure patients, alleviating clinical symptoms, reducing left ventricular end-diastolic diameter, increasing LVEF, increasing the achievement rate of clinical target doses of neuroendocrine system antagonists (β-blockers and ACEI / ARB / ARNI), and helping to follow guideline-directed medical therapy (GDMT); 2. The Qi-invigorating and Yang-tonifying formula can significantly inhibit the thickness of the interventricular septum during diastole and systole, the thickness of the left ventricular posterior wall during diastole and systole in DOX-HF mice, increase the left ventricular ejection fraction in mice, effectively alleviate the changes in cardiomyocyte morphology and cardiac fibrosis caused by DOX; it can correct the reduction in acylcarnitine content caused by DOX, and can reverse the inhibition of fatty acid β-oxidation function of cardiac tissue by DOX, thereby reducing cardiac damage caused by anthracycline chemotherapy drugs. This provides a pharmacological and molecular biological basis for the Yiqi Wenyang Formula to improve heart failure caused by anthracycline chemotherapy drugs; 3. Five active ingredients were identified in the water extract of Yiqi Wenyang Formula by LC-MS / MS analysis, namely, gentianin, verbascoside-7-O-β-D-glucopyranoside, astragaloside IV, rhodioloside, and paeoniflorin. Attached Figure Description

[0022] Figure 1The effects of Yiqi Qiyang formula and SR9009 on echocardiographic parameters in mice treated with DOX: A: echocardiograms of each group, B: left ventricular ejection fraction (EF%) of each group, C: left ventricular shortening fraction (FS%) of each group, D: diastolic interventricular septal thickness (IVS d) of each group, E: systolic interventricular septal thickness (IVS s) of each group, F: left ventricular end-diastolic anterior wall diameter (LVID d) of each group, G: left ventricular end-systolic anterior wall diameter (LVID s) of each group, H: diastolic left ventricular posterior wall thickness (LVPW d) of each group, I: statistical data on systolic left ventricular posterior wall thickness (LVPW s) of each group;

[0023] Figure 2 The effects of Yiqi Qiyang formula and SR9009 on HW / BW and HW / TL in DOX-HF mice: A: Effects of Yiqi Qiyang formula and SR9009 on HW / BW in DOX-HF mice; B: Effects of Yiqi Qiyang formula and SR9009 on HW / TL in DOX-HF mice;

[0024] Figure 3 Effects of Yiqi Qiyang formula and SR9009 on the morphology and cardiac fibrosis of DOX-treated mouse cardiomyocytes: A: HE staining images of mouse cardiomyocytes in each group (100X); B: HE staining images of mouse cardiomyocytes in each group (400X); C: Sirius staining images of mouse cardiomyocytes in each group (100X); D: Sirius staining images of mouse cardiomyocytes in each group (200X);

[0025] Figure 4 The results are from PLS-DA analysis in positive ion mode.

[0026] Figure 5 The results are from PLS-DA analysis in negative ion mode.

[0027] Figure 6 The positive ion mode thermal image is used to display the identified substances;

[0028] Figure 7 The thermal image of the negative ion mode is used to identify the substances.

[0029] Figure 8 Heatmap of differential expression of all identified acylcarnitines;

[0030] Figure 9 Figure 1 shows the fatty acid metabolism of the heart in DOX-HF mice: A: Representative PET / CT images of mice in the Control group; B: Representative PET / CT images of mice in the DOX intervention group; C: Representative PET / CT images of mice in the DOX + Yiqi Wenyang Formula intervention group; D: Statistical analysis results of myocardial %ID / g values ​​in each group of mice; E: Left ventricular %ID / g values ​​in each group of mice; F: Statistical analysis results of whole cardiac %ID / g values ​​in each group of mice. Detailed Implementation

[0031] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0032] All the Chinese herbs in the Yiqi Qiyang formula were provided by the Affiliated Hospital of Nanjing University of Chinese Medicine.

[0033] Example 1: Preparation of Qi-invigorating and Yang-tonifying freeze-dried powder

[0034] 1. The formula for Yiqi Qiyang Decoction consists of 60g of raw Astragalus membranaceus, 20g of Rhodiola rosea, 6g of processed Aconitum carmichaelii, 10g of Polyporus umbellatus, 10g of Lepidium apetalum, 10g of Curcuma zedoaria, 12g of Paeonia lactiflora, and 3g of dried ginger. Decoction preparation: To ensure consistency with clinical dosage, Yiqi Qiyang Decoction is extracted using traditional decoction methods. Each dose of herbs is soaked in 1.31L of water for 30 minutes, brought to a boil over high heat, then simmered over low heat for 1 hour, collecting the first decoction. The residue is then added to 1.048L of cold water, brought to a boil again, then simmered over low heat for 1 hour, collecting the second decoction. The two decoctions are combined and heated to concentrate to half their original volume. The concentrate is then poured out and coarsely filtered through a double-layered gauze to obtain the decoction. This decoction is then placed in a freeze dryer for vacuum freeze-drying to obtain the freeze-dried powder of Yiqi Qiyang Decoction. In this formula, raw astragalus tonifies qi and promotes diuresis, serving as the principal herb; rhodiola rosea tonifies qi, nourishes the heart, and invigorates blood; and prepared aconite root warms and tonifies the heart and kidneys, serving as the assistant herbs; polyporus umbellatus and lepidium seed purge the lungs and promote diuresis; and turmeric root breaks up blood stasis and promotes qi circulation, serving as adjuvant herbs; and white peony root and dried ginger serve as guiding herbs. The entire formula works to tonify qi and warm yang, resolve blood stasis, and promote diuresis. Based on tonifying heart qi and warming heart yang, it further uses polyporus umbellatus and lepidium seed to promote diuresis, and turmeric root to promote qi circulation and break up blood stasis, ensuring that tonification does not lead to stagnation, achieving a balance of clearing and tonifying, and mildly draining, resulting in twice the effect with half the effort.

[0035] 2. LC-MS / MS detection of Yiqi Qiyang formula

[0036] Follow the article (YQWY decoction reverses cardiac hypertrophy induced by TACthrough inhibiting GATA4 phosphorylation and MAPKs HUANG Jing-Jing1Δ,XIEYong1Δ,LI He1,ZHANG Xiao-Xiao1,HUANG Qing1,ZHU Yao1,GU Ping2*,JIANG The method described in Wei-Min1*) was used to perform LC-MS / MS detection on the water extract (the freeze-dried powder of Yiqi Wenyang Fang dissolved in water). The LC-MS / MS results showed that five compounds were identified in the water extract: mangiferin (RT = 1.630 min), verbascoside-7-O-β-D-glucopyranoside (RT = 4.299 min), astragaloside IV (RT = 9.106 min), rhodioloside (RT = 9.383 min), and paeoniflorin (RT = 13.175 min), with contents of 0.348 μg / mg, 0.289 μg / mg, 0.317 μg / mg, 0.258 μg / mg, and 0.643 μg / mg, respectively. Among them, 1: Formononetin; 2: Calycosin; 3: Astragaloside IV; 4: Salidroside; 5: Paeoniflorin.

[0037] Example 2: Regulation of cardiac ultrasound parameters in DOX-HF mice by the Qi-invigorating and Yang-tonifying formula

[0038] Animal source and grouping: Eight-week-old male C57BL / 6 mice were used as the research subjects. All mice were purchased from Beijing Vital River Co., Ltd. All mice were housed in the Animal Center of Nanjing University of Chinese Medicine in an SPF-grade laboratory with a 12-hour diurnal cycle and constant temperature and humidity (temperature 23±1℃, humidity 60%). A DOX-HF model was established. Eight-week-old male mice were randomly divided into five groups: control group, DOX modeling group, low-yield Qi Yang Fang group, medium-yield Qi Yang Fang group, and high-yield Qi Yang Fang group, with 10 mice in each group. The model was established by administering four doses of DOX, each at a dose of 6 mg / kg to mice, every two days. The DOX solution was diluted with PBS on the day of injection, resulting in a concentration of 600 ug / ml. The solution was sterilized using a 0.22 μm filter membrane and administered via intraperitoneal injection on days 8, 11, 14, and 17 after the start of the experiment, for a total of four injections, totaling 24 mg / kg (Reference: Fang Yuan. The intervention effect of Qianyang Yuyin Granules on the regulation of cardiac and renal fibrosis based on the TGF-β1 signaling pathway [D]. Nanjing University of Chinese Medicine, 2019). The low-yiqi-qi-yang formula group, medium-yiqi-qi-yang formula group, and high-yiqi-qi-yang formula group were given lyophilized powder solutions of the yiqi-qi-yang formula decoction (wherein, the lyophilized powder production rate: crude drug: lyophilized powder = 1:0.1104; the gavage volume for mice was 0.2 ml / 10 g; the equivalent dose group lyophilized powder solution: 323 mg / ml; the low-dose group lyophilized powder solution: 161.5 mg / ml; the high-dose group lyophilized powder solution: 646 mg / ml). The medium-dose group received the equivalent human dose of crude drug (29.27 g crude drug / kg / d), the low-dose group received half the medium dose (14.635 g crude drug / kg / d), and the high-dose group received twice the medium dose (58.54 g crude drug / kg / d). The equivalent dose was calculated using the human and mouse body surface area formula. During gavage, the aqueous extract of the Qi-tonifying and Yang-warming formula was weighed according to the mass ratio of crude drug to raw drug (crude drug: lyophilized powder = 1:0.1104), dissolved in water, and then administered by gavage. Gavage with the traditional Chinese medicine began simultaneously with DOX modeling, once daily at ZT6 (ZT0 is the timing factor, i.e., the time the lights were turned on. For example, ZT0 is defined as 7:00 AM, and ZT6 is 1:00 PM), for 18 consecutive days. On day 26 of the experiment, mice were euthanized with an excessive dose of sodium pentobarbital (250 mg / kg, ip). Blood samples were centrifuged at 3500 rpm for 10 minutes to obtain serum, which was stored at -80°C for further analysis. After blood collection, heart tissue was removed, weighed, and its mass recorded and stored at -80°C for further analysis. Additionally, tibial length was measured after dissecting skeletal muscle and soft tissue to expose the tibia. These animal experiments complied with the National Institutes of Health's guidelines for the use of laboratory animals. All experiments were approved by the Animal Ethics Committee of the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine (Nanjing, Jiangsu).

[0039] Methods: Echocardiography was performed on day 25 of the experiment at the ultrasound laboratory of the Jiangsu Provincial Medical Animal Experiment Base using a vevo 2100 high-frequency color Doppler ultrasound system with a 30MHz center frequency probe. Image acquisition was performed using M-mode and two-dimensional images at the parasternal papillary muscle level. The parameters examined in this study included IVS d, IVS s, LVID d, LVID s, LVPW d, LVPW s, and LVFS (FS%).

[0040] The results are as follows Figure 1As shown, the control group was the saline intervention group, the DOX group was the DOX intervention group, the YQWY group (medium-dose warming and tonifying formula) was the DOX+YQWY intervention group, and the SR9009 group (activator of the circadian rhythm gene Nr1d1) was the DOX+SR9009 intervention group. Data are expressed as mean ± standard deviation. *P<0.05, **P<0.01, relative to the control group. #P<0.05, ##P<0.01, relative to the DOX group. Control group: saline intervention group (n=12), DOX group (n=16), YQWY group (n=16), SR9009 group (n=16). Where n is the number of animals. Compared with the saline group, the DOX group mice showed the following changes: left ventricular ejection fraction (LVEF, EF%, 63.06±2.007 vs 46.83±4.995), left ventricular shortening fraction (FS%, 33.53±1.392 vs 23.11±2.898), diastolic interventricular septal thickness (IVS d, 0.7491±0.02503 vs 0.6149±0.06440), systolic interventricular septal thickness (IVS s, 1.164±0.04514 vs 0.8841±0.08497), diastolic left ventricular posterior wall thickness (LVPW d, 0.7548±0.03605 vs 0.6109±0.05764), and systolic left ventricular posterior wall thickness (LVPW s, 1.277±0.08683 vs 0.6109±0.05764). The values ​​of EF% (46.83±4.995 vs 58.74±3.764), FS% (23.11±2.898 vs 30.51±2.504), IVS d (0.6149±0.06440 vs 0.6865±0.04226), IVS s (0.8841±0.08497 vs 1.050±0.07308), LVPW d (0.6109±0.05764 vs 0.6776±0.03506), and LVPW s (0.8736±0.09957 vs 1.095±0.07453) were significantly lower in the Yiqi Wenyang group (YQWY).Compared with the DOX group, the SR9009 group also showed significant improvements in EF% (46.83±4.995 vs 58.57±4.226), FS% (23.11±2.898 vs 30.42±2.802), IVS d (0.6149±0.06440 vs 0.6593±0.05124), IVS s (0.8841±0.08497 vs 1.007±0.09663), LVPW d (0.6109±0.05764 vs 0.6698±0.05962), and LVPW s (0.8736±0.09957 vs 1.061±0.1139). Figure 1 ).like Figure 1 As shown, compared with the saline group, the left ventricular end-systolic anterior wall diameter (LVIDs, 2.492±0.1972 vs 3.003±0.1838) was significantly increased in the DOX group. Compared with the DOX group, the LVIDs of the Qi-tonifying and Yang-warming group (3.003±0.1838 vs 2.586±0.2766) and the SR9009 group (3.003±0.1838 vs 2.587±0.1831) were also significantly decreased. There was no statistically significant difference in LVIDs among the three groups.

[0041] Example 3: Effects of the Qi-invigorating and Yang-tonifying formula on HW / BW and HW / TL in DOX-HF mice

[0042] Methods: See the "Animal Source and Grouping" section.

[0043] like Figure 2As shown, compared with the saline group, DOX increased heart weight / body weight (4.837±0.2141 vs 6.378±0.6708, HW / BW, mg / g) and heart weight / tibia length (7.029±0.5342 vs 7.748±0.8287, HW / TL, mg / mm) in mice. Meanwhile, a medium dose of Yiqi Wenyang formula decreased HW / BW (6.378±0.6708 vs 5.490±0.3114, mg / g) and HW / TL (7.748±0.8287 vs 7.078±0.5317, mg / mm) in DOX-HF mice. SR9009 also decreased HW / BW (6.378±0.6708 vs 5.490±0.3114, mg / g) in DOX-HF mice. 5.026±0.6546 (mg / g), HW / TL (7.748±0.8287 vs 7.250±0.6168, mg / mm). It is evident that the Qi-tonifying and Yang-warming formula can reduce HW / BW and HW / TL in DOX-HF mice. *P<0.05, **P<0.01, relative to Control. #P<0.05, ##P<0.01, relative to DOX. Control: saline intervention group (n=16), DOX: DOX intervention group (n=16), YQWY: DOX+YQWY intervention group (n=16), SR9009: DOX+SR9009 intervention group (n=16). Data are presented as mean ± standard deviation.

[0044] Example 4: Effects of the Qi-invigorating and Yang-tonifying formula on the cardiac pathology of DOX-HF mice

[0045] (1) Heart tissues from each group of mice were isolated, and HE staining was used to evaluate the morphology of cardiomyocytes. The specific method was as follows: Heart tissues were fixed in 4% paraformaldehyde solution, embedded in paraffin, and cut into sections with a thickness of about 5 μm. The sections were first soaked in xylene solution for 20 minutes, then soaked in fresh xylene solution (purchased from Sinopharm Chemical Reagent Co., Ltd., ≥99.0%) for 20 minutes, then soaked in pure alcohol for 5 minutes, then soaked in fresh pure alcohol for 5 minutes, and then soaked in 75% ethanol for 5 minutes. The sections were then rinsed with pure water to complete the dewaxing. The sections were then soaked in hematoxylin staining solution for 5 minutes, rinsed with pure water, differentiated with differentiation solution, rinsed with pure water, blued with blue solution, and rinsed with pure water. The sections were then sequentially immersed in 85% and 95% graded alcohol for 5 minutes each, and then incubated in eosin solution for 5 minutes. Finally, the sections were incubated in pure alcohol for 5 minutes each, for a total of 3 incubations, and then incubated in xylene for 5 minutes x 2 times. After mounting with neutral resin, the sections were observed under a microscope.

[0046] (2) Heart tissues from each group of mice were isolated, and the degree of cardiac fibrosis was assessed using Sirius red staining. The specific method was as follows: Heart tissues were fixed in 4% paraformaldehyde solution, embedded in paraffin, and cut into sections approximately 5 μm thick. The paraffin sections were dewaxed, rinsed with tap water, and then incubated overnight with potassium dichromate solution. They were then stained with iron hematoxylin solution for 5 minutes, rinsed with tap water, and then differentiated with 1% hydrochloric acid ethanol. After rinsing with tap water, they were incubated with virgin red and acid fuchsin solution for 5 minutes. After a brief rinse with tap water, they were incubated with 1% phosphomolybdic acid solution for 3 minutes. After incubation, they were directly incubated with aniline blue solution for 5 minutes. Afterward, without rinsing with tap water, they were directly incubated in 1% glacial acetic acid for 1 minute. After rapid washing with water, they were dehydrated, mounted, and then photographed under a microscope.

[0047] The results show ( Figure 3 Medium doses of Yiqi Qiyang formula and SR9009 can effectively alleviate DOX-induced changes in myocardial cell morphology and cardiac fibrosis.

[0048] Effects of medium-dose Qi-invigorating and Yang-tonifying formula on the metabolomics of cardiac tissue in DOX-HF mice in Example 5

[0049] To explore whether changes in fatty acid β-oxidation genes related to DOX action and the regulation of YQWY affect changes in the overall metabolite levels of the animal heart, this invention performed non-target metabolomics analysis on DOX-HF mice.

[0050] Instruments: Mass spectrometry: Thermo QE HF-X mass spectrometer; Chromatography: Thermo Vanquish liquid chromatograph; Chromatographic column: Waters, ACQUITY UPLC HSS T3 1.7μm, 2.1mm×150mm column.

[0051] Reagents: Acetonitrile (Merck), acetic acid (Sigma), methanol (Merck), formic acid (Merck).

[0052] The sample was derived from Example 2.

[0053] 1. Metabolite extraction

[0054] Remove the sample from the -80℃ freezer and place it on prepared dry ice. Add 400 μL of extraction buffer (methanol:acetonitrile = 3:1, pre-cooled at -40℃), vortex for 5 min, sonicate for 15 min, and let stand at 4℃ for 2 hours. Centrifuge the sample for 15 min (12000 rpm, 4℃), take an equal volume, and concentrate to dryness under vacuum. Redissolve in 100 μL of 50% methanol aqueous solution (methanol:water = 1:1, v / v), vortex for 3 min (4℃, 2000 rpm), centrifuge for 15 min (12000 rpm, 4℃), and use the supernatant for analysis.

[0055] 2. Chromatographic separation

[0056] Throughout the analysis, the sample was placed in an 8°C autosampler and separated using an HSST3 column in an ultra-high performance liquid chromatography (UHPLC) system. The injection volume was 2 μL, the column temperature was 40°C, and the flow rate was 0.3 mL / min. The mobile phase consisted of A: 0.1% formic acid in methanol and B: 0.05% acetic acid in methanol. The gradient elution program was as follows: positive ion: 0-0.5 min, A: 2%; 0.5-6 min, A: 2-50%; 6-10 min, A: 50-98%; 10-14 min, A: 98%; 14-16 min, A: 98-2%; 16-21 min, A: 2%. Negative ion concentrations: 0-0.5 min, B: 2%; 0.5-6 min, B: 2-50%; 6-10 min, B: 50-98%; 10-14 min, B: 98%; 14-16 min, B: 98-2%; 16-21 min, B: 2%. QC samples were inserted into the sample queue to monitor and evaluate the system's stability and the reliability of the experimental data.

[0057] 3. Mass spectrometry acquisition

[0058] Each sample was detected using electrospray ionization (ESI) in both positive and negative ion modes. After separation by UHPLC, samples were analyzed by mass spectrometry using a Thermo QE HF-X mass spectrometer. Mass spectrometry conditions: Ionization source: ESI; sheath gas flow rate: 30; auxiliary gas: 10; spray voltage: 2.5 kV(+) / 2.5 kV(-); S-Lens RF: 50; capillary temperature: 325 °C; auxiliary gas temperature: 300 °C; second-order collision energy (NCE): 30; isolation window: 1.5 m / z; Top N = 8. Scan range: 70-1050 m / z; scanning mode: separate scans for positive and negative ions. Analytical software: Xcalibur version 4.1.

[0059] Results: Both positive ion mode and negative ion mode collected response peaks, which could reflect the typical differences among the three groups. Figure 4 , Figure 5 After peak alignment, noise filtering, and database matching using MS-DAIL software, a total of 2409 metabolites were identified in positive ion mode and 1607 in negative ion mode. Figure 6 , Figure 7Among these substances, the contents of acylcarnitines—Acetyl-L-carnitine, Stearoyl-L-carnitine (C18:0), L-Palmitoylcarnitine (C16 Carnitine), Tetradecanoyl-L-carnitine (C14 Carnitine), Octanoyl-L-carnitine (C8 Carnitine), and Hexanoyl-L-carnitine (C6 Carnitine)—were significantly decreased in the DOX group and significantly increased in the YQWY group. Figure 8 , (cutoff: P value<0.05, |logFC|>0.5, VIP>1)).

[0060] The effect of medium-dose Qi-tonifying and Yang-warming formula on fatty acid metabolism in the heart of DOX-HF mice in Example 6

[0061] In this embodiment, 18F-FTHA PET / CT was used to observe the metabolism of fatty acids in the hearts of mice in each group.

[0062] Mice from the DOX group, DOX+YQWY intervention group, and saline intervention group constructed in Example 2 were sent to Mido (Nanjing) Biotechnology Co., Ltd. for PET-CT examination.

[0063] Result: As Figure 9 As shown, the groups were: Control: saline intervention group (n=3), DOX: DOX intervention group (n=3), and YQWY: DOX+YQWY intervention group (n=3). Figure A shows representative images of mice in the Control group, Figure B shows representative images of mice in the DOX group, Figure C shows representative images of mice in the YQWY group, Figure D shows the statistical analysis results of myocardial %ID / g between groups, Figure E shows the statistical analysis results of left ventricular %ID / g, and Figure F shows the statistical analysis results of whole heart %ID / g. Compared with the saline group, the whole heart %ID / g value of mice in the DOX group was significantly decreased, and the difference was statistically significant (P<0.05), while the changes in myocardial %ID / g and left ventricular %ID / g values ​​were not statistically significant (P>0.05). Compared with the DOX group mice, the myocardial %ID / g and whole-heart %ID / g values ​​of the YQWY group mice were significantly increased (P<0.05). The left ventricular %ID / g value showed an increasing trend, but the difference was not statistically significant (P>0.05). Data are expressed as mean ± standard deviation. *P<0.05, relative to Control. #P<0.05, relative to DOX.

[0064] Conclusion: DOX mice showed reduced fatty acid metabolism in their hearts, while YQWY could improve fatty acid metabolism.

Claims

1. A traditional Chinese medicine composition for invigorating qi and warming yang, characterized in that, The ingredients are made from the following raw materials in parts by weight: 50-70 parts raw Astragalus membranaceus, 10-30 parts Rhodiola rosea, 3-8 parts processed Aconitum carmichaelii, 5-15 parts Polyporus umbellatus, 5-15 parts Lepidium apetalum, 5-15 parts Curcuma zedoaria, 5-18 parts Paeonia lactiflora, and 1-7 parts dried ginger.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients are made from the following raw materials in parts by weight: 55-65 parts raw Astragalus membranaceus, 15-25 parts Rhodiola rosea, 3-6 parts processed Aconitum carmichaelii, 10-15 parts Polyporus umbellatus, 10-15 parts Lepidium apetalum, 10-15 parts Curcuma zedoaria, 10-18 parts Paeonia lactiflora, and 1-5 parts dried ginger.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The ingredients are made from the following raw materials in parts by weight: 60 parts raw Astragalus membranaceus, 20 parts Rhodiola rosea, 6 parts processed Aconitum carmichaelii, 10 parts Polyporus umbellatus, 10 parts Lepidium apetalum, 10 parts Curcuma zedoaria, 12 parts Paeonia lactiflora, and 3 parts dried ginger.

4. A traditional Chinese medicine preparation, characterized in that, Includes the traditional Chinese medicine composition as described in claim 1.

5. The traditional Chinese medicine preparation according to claim 4, characterized in that, It also includes pharmaceutically acceptable excipients.

6. A freeze-dried powder containing the traditional Chinese medicine composition for invigorating qi and warming yang as described in claim 1.

7. A method for preparing the freeze-dried powder according to claim 6, characterized in that, Soak the herbal composition according to any one of claims 1 to 3 in water, bring to a boil over high heat, simmer over low heat, and collect the first decoction; add cold water to the dregs, continue to boil, then simmer over low heat, and collect the second decoction; mix the two decoctions and continue to heat and concentrate to half of the original volume, pour out and coarsely filter with a double-layer gauze to obtain the herbal decoction; place the above decoction in a freeze dryer for vacuum freeze drying to obtain the freeze-dried powder of Yiqi Qiyang formula.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3, the traditional Chinese medicine preparation according to any one of claims 4 to 5, or the freeze-dried powder according to claim 6 in the preparation of a drug for treating pharmacological heart failure.

9. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3, the traditional Chinese medicine preparation according to any one of claims 4 to 5, or the lyophilized powder according to claim 6 in the preparation of a medicament for treating cardiac damage and / or heart failure caused by anthracycline chemotherapy drugs.

10. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3, the traditional Chinese medicine preparation according to any one of claims 4 to 5, or the lyophilized powder according to claim 6 in the preparation of a medicament for treating cardiac damage and / or heart failure caused by doxorubicin.

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