A traditional Chinese medicine composition for preventing and treating myocardial fibrosis and its medical use

Abstract

The main purpose of the present invention is to provide a traditional Chinese medicine composition for the prevention and treatment of myocardial fibrosis. The traditional Chinese medicine composition is mainly prepared from leeches, Chuanxiong rhizome, astragalus root, and salvia miltiorrhiza. The traditional Chinese medicine composition of the present invention can inhibit the formation and progression of myocardial fibrosis in rats by inhibiting the increase in type I collagen synthesis induced by isoproterenol. The traditional Chinese medicine composition of the present invention can treat myocardial fibrosis in spontaneously hypertensive rats (SHR). The traditional Chinese medicine composition of the present invention is effective in preventing and treating myocardial fibrosis and has comprehensive effects.

Description

Technical Field

[0001] The present invention relates to a composition and a preparation method thereof, in particular to a traditional Chinese medicine composition for preventing and treating myocardial fibrosis and medical use thereof, belonging to the field of medicine. Background Art

[0002] Myocardial fibrosis (MF) is a common pathological change in various heart diseases at certain stages of development. It is one of the most characteristic structural changes and a major manifestation of ventricular remodeling after MI, which can promote myocardial systolic and diastolic dysfunction. Studies have shown that long-term activation of cardiac fibroblasts (CFs), characterized by increased collagen content, myofibroblast differentiation, and concomitant extracellular matrix secretion, can lead to myocardial fibrosis.

[0003] Cardiomyocytes are highly differentiated terminal cells. Under pathological conditions such as chronic mechanical stimulation, humoral stimulation, pressure, and / or excessive volume overload, cardiac fibroblasts proliferate and transform into myofibroblasts, leading to increased synthesis and decreased degradation of extracellular matrix (ECM) proteins such as collagen. The content of these ECM components can be used to assess the degree of myocardial fibrosis. Type I and type III collagen are the major components of the ECM, accounting for 80% and 11% of myocardial collagen, respectively. As myocardial fibrosis progresses, myocardial stiffness increases and compliance decreases, leading to restricted filling and diastolic dysfunction. This directly activates the RAAS system, causing sodium and water retention, further stimulating the atria to secrete atrial natriuretic factor (ANF) to dilute sodium and water. ANF is a key clinical indicator of cardiac function.

[0004] Currently, the main clinical drug treatments for myocardial fibrosis include angiotensin-converting enzyme inhibitors (ACEIs), aldosterone receptor antagonists (ARBs), and beta-blockers. However, because ACEIs can cause irritating dry coughs and angioedema, many patients cannot tolerate them. The incidence of hyperkalemia with ARBs is as high as 8.6% to 26%, often with arrhythmias as the first manifestation. Blood potassium and electrocardiograms must be closely monitored during medication. Beta-blockers carry the risk of causing hypotension, inducing or exacerbating bronchial asthma, and worsening atrioventricular block. Furthermore, sudden discontinuation of long-term use often exacerbates the original symptoms.

[0005] Chuanzhi Tongluo Capsules are used to treat blood stasis and qi deficiency in the meridians (cerebral infarction) during the recovery phase of stroke. Symptoms include hemiplegia, slurred speech or loss of speech, hemiplegic numbness, facial paralysis, fatigue, a dark tongue with ecchymosis or petechiae, visible sublingual veins, a deep, thready, or stringy pulse, and a thin, white fur. Chuanzhi Tongluo Capsules are formulated with four traditional Chinese medicinal herbs: leech, Chuanxiong rhizome, astragalus, and salvia miltiorrhiza. Developed based on Buyang Huanwu Decoction, Chuanzhi Tongluo Capsules are formulated with leech, Chuanxiong rhizome, astragalus, and salvia miltiorrhiza.

[0006] Wang Fei et al. (Wang Fei, Rao Feng, Qin Yaqin, Zhao Fulan. Efficacy of Chuanzhi Tongluo Capsule combined with rosuvastatin in patients with normal blood lipids and acute cerebral infarction [J]. World Chinese Medicine, 2019, 14(04): 887-890) studied the efficacy of Chuanzhi Tongluo Capsule combined with rosuvastatin in patients with normal blood lipids and acute cerebral infarction, and found that the efficacy of Chuanzhi Tongluo Capsule combined with rosuvastatin in the treatment of normal blood lipids in ACI was significantly better than that of rosuvastatin alone. Wu Qingqiang et al. (Wu Qingqiang, Lin Yafa, Wang Yonghe. Clinical observation of Chuanzhi Tongluo Capsule combined with cefuroxime in the treatment of adult lobar pneumonia [J]. Pharmacology and Clinic of Traditional Chinese Medicine, 2017, 33(01): 175-177) found that the inflammation level of patients was better controlled after the combined use of Chuanzhi Tongluo Capsule. It is speculated that the reason may be related to the antibacterial activity of multiple Chinese medicinal ingredients in Chuanzhi Tongluo Capsule. Summary of the Invention

[0007] The main purpose of the present invention is to provide a traditional Chinese medicine composition for preventing and treating myocardial fibrosis. The traditional Chinese medicine composition is mainly prepared from leech, chuanxiong, astragalus and salvia miltiorrhiza.

[0008] One of the purposes of the present invention is to provide the composition of the above-mentioned Chinese medicine composition, that is, the Chinese medicine composition is mainly prepared from the following raw materials:

[0009] 300-750 parts by weight of leech, 300-750 parts by weight of chuanxiong, 250-500 parts by weight of salvia miltiorrhiza, and 250-500 parts by weight of astragalus.

[0010] Preferably, the Chinese medicine composition is mainly prepared from the following raw materials:

[0011] 450-700 parts by weight of leech, 450-700 parts by weight of chuanxiong, 300-450 parts by weight of salvia miltiorrhiza, and 300-450 parts by weight of astragalus.

[0012] In one of the further preferred solutions, the Chinese medicine composition is mainly prepared from the following raw materials:

[0013] 600 parts by weight of leech, 600 parts by weight of chuanxiong, 400 parts by weight of salvia miltiorrhiza, and 400 parts by weight of astragalus.

[0014] The second object of the present invention is to provide specific uses of the above-mentioned Chinese medicine composition, mainly including:

[0015] The present invention found through animal experiments that the type I collagen content in the serum, the hydroxyproline content in the myocardial tissue, and the left ventricular index in each group of rats that were gavage-administered with the traditional Chinese medicine composition of the present invention were significantly reduced, with significant differences (P < 0.05) or extremely significant differences (P < 0.01) compared with the model control group. This indicates that the traditional Chinese medicine composition of the present invention can inhibit the increase in type I collagen synthesis induced by isoproterenol, thereby inhibiting the formation and development of myocardial fibrosis in rats.

[0016] The present invention studies the influence of the traditional Chinese medicine composition on myocardial fibrosis in spontaneously hypertensive rats (SHR), and confirms that the traditional Chinese medicine composition of the present invention has the effect of resisting myocardial fibrosis.

[0017] In the Chuanzhi Tongluo Capsule, leech is the main medicine, which is good at resolving blood stasis, breaking up blood and eliminating accumulation; Chuanxiong is the auxiliary medicine, which has an upward nature and can cooperate with leech to reach the brain meridians, promote blood circulation and dredge the meridians, so that the overall qi and blood flow smoothly and the functions of the internal organs are strong; astragalus and salvia miltiorrhiza are adjuvants. Astragalus replenishes qi to relieve blood stasis, warm and dredge the meridians, and alone it promotes blood circulation, removes blood stasis, calms the mind and nourishes the heart; the whole formula has the effects of promoting blood circulation, removing blood stasis, invigorating qi and dredging the meridians.

[0018] The third object of the present invention is to provide a Chinese medicine preparation containing the above-mentioned Chinese medicine composition, wherein the Chinese medicine preparation is a clinically acceptable oral preparation;

[0019] Preferably, the clinically acceptable oral preparation is one or more of a pill, a capsule, a tablet, a granule or a liquid oral preparation;

[0020] More preferably, the oral preparation is a capsule.

[0021] Compared with the prior art, the present invention has achieved remarkable technical effects:

[0022] The Chinese herbal composition of the present invention is effective and comprehensive in preventing and treating myocardial fibrosis. Clinical results have shown that the Chinese herbal composition of the present invention has low toxicity and side effects. The discovery of this new use for preventing and treating myocardial fibrosis has broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 HE staining and PAS staining of the Chinese medicinal composition of Example 1 for preventing and treating myocardial fibrosis. Specific embodiments

[0024] Formula Example 1

[0025] 600 parts by weight of leech, 600 parts by weight of chuanxiong, 400 parts by weight of salvia miltiorrhiza, and 400 parts by weight of astragalus.

[0026] Prescription Example 2

[0027] 300 parts by weight of leech, 750 parts by weight of chuanxiong, 250 parts by weight of salvia miltiorrhiza, and 500 parts by weight of astragalus.

[0028] Formula Example 3

[0029] 750 parts by weight of leech, 300 parts by weight of chuanxiong, 500 parts by weight of salvia miltiorrhiza, and 250 parts by weight of astragalus.

[0030] Prescription Example 4

[0031] 700 parts by weight of leech, 450 parts by weight of chuanxiong, 450 parts by weight of salvia miltiorrhiza, and 300 parts by weight of astragalus.

[0032] Prescription Example 5

[0033] 450 parts by weight of leech, 700 parts by weight of chuanxiong, 300 parts by weight of salvia miltiorrhiza, and 450 parts by weight of astragalus.

[0034] Example 1 Study on the efficacy of the Chinese medicine composition of the present invention in preventing and treating myocardial fibrosis

[0035] Isoproterenol, a β1 receptor agonist, has been shown to induce myocardial fibrosis in rats through continuous administration, primarily by increasing Angll concentrations in the circulatory system and myocardium. This model has become a common method for studying myocardial fibrosis due to its simplicity, low cost, and long-lasting effects.

[0036] Forty SD rats, half female and half female, weighing 180-220 g, were randomly divided into the following five groups: normal group, model group, low-dose Chinese medicine group, medium-dose Chinese medicine group, and high-dose Chinese medicine group, with 8 rats in each group.

[0037] Except for the normal group, all other groups received a subcutaneous injection of isoproterenol 5 mg / (kg·d) on the back for 7 consecutive days. The rats in each Chinese medicine group were given Chuanzhi Tongluo Capsule by gavage starting from the second day for 14 days, once a day, with a gavage dose of 10 mL / kg each time. The normal group and the model group were given the same volume of purified water every day.

[0038] Chuanzhi Tongluo Capsule: Lunan Houpu Pharmaceutical Co., Ltd.

[0039] Low-dose Chinese medicine group: 60 mg / kg Chuanzhi Tongluo capsule;

[0040] Medium dose group of traditional Chinese medicine: 120mg / kg Chuanzhi Tongluo capsule;

[0041] High-dose Chinese medicine group: 240 mg / kg Chuanzhi Tongluo capsule;

[0042] After the last dose, the rats were fasted but not watered for 24 h. After being weighed, they were anesthetized with 10% chloral hydrate (0.3 mL / kg). Blood was collected from the abdominal aorta and serum was collected for later use. The heart was removed by thoracotomy, and the large blood vessels and epicardial fat tissue were removed. The heart was washed with physiological saline, dried, and weighed to calculate the left ventricular weight index (left ventricular weight / body weight).

[0043] Serum was collected and type I collagen content was measured using an ELISA kit using a microplate reader according to the kit's instructions. Additionally, after rinsing the heart tissue with ice-cold saline, 150 mg of left ventricular myocardial tissue was collected and subjected to alkaline hydrolysis. Hydroxyproline content in the myocardial tissue was measured spectrophotometrically at a wavelength of 550 nm according to the kit's instructions. The test results for these indicators are shown in Table 1.

[0044] Table 1 Effects of the Chinese medicine composition of the present invention on various indicators of rats with myocardial fibrosis

[0045]

[0046] Compared with the normal group # P<0.05, # P < 0.01;

[0047] Compared with the model group, *p<0.05, **p<0.01.

[0048] The experimental results in Table 1 show that the serum type I collagen content, myocardial tissue hydroxyproline content, and left ventricular index in the model group were significantly higher than those in the control group. The present invention uses isoproterenol to induce a rat myocardial fibrosis model, which is manifested by an increase in the left ventricular index, an increase in the left ventricular hydroxyproline content, and an increase in the serum type I collagen content, indicating that the myocardial fibrosis model is established.

[0049] The results of the Chinese herbal medicine combination on the pathological changes of myocardial fibrosis rats are shown in Figure 1 . Figure 1 The results showed that the rats in the model group had myocardial cell degeneration and necrosis, interstitial inflammatory cell infiltration and fibrous tissue proliferation; after administration of the Chinese medicine composition of the present invention, the pathological changes were significantly improved. The Chinese medicine composition of the present invention can improve myocardial fibrosis and remodeling.

[0050] The serum type I collagen content, myocardial tissue hydroxyproline content, and left ventricular index of each group of the Chinese medicine composition of the present invention were significantly reduced, with significant or extremely significant differences compared with the model control group. This indicates that the Chinese medicine composition of the present invention can inhibit the increase in type I collagen synthesis induced by isoproterenol, thereby inhibiting the formation and development of myocardial fibrosis in rats.

[0051] Example 2 Effect of the Chinese medicine composition of the present invention on myocardial fibrosis in SHR rats

[0052] The present invention uses spontaneously hypertensive rats (SHR) as an experimental animal model and is conducted according to the method of the following document (Kuijpers M HM, Jong W D. Spontaneous hypertension in the fawn-hooded rat: a cardiovascular disease model [J]. J Hypertens Suppl, 1986, 4(3): 41-4.). The method proposes that fawn-hooded rats can develop spontaneous hypertension at 5 weeks of age, which increases with age, reaching 180-240 mmHg at 1 year of age, accompanied by increased weight of organs such as the heart, kidneys, liver, and spleen, and histopathological changes such as myocardial fibrosis and myocardial infarction. Since then, many scholars have applied SHR to the study of hypertension-induced myocardial fibrosis, such as the literature (Barone FC, Campbell WG, Nelson AH, et al. Carvedilol prevents severe hypertensive cardiomyopathy and remodeling [J]. Journal of Hypertension, 1998, 16(6): 871., and Huikeshoven M, Wal A CV D, Tukkie R, et al. The old spontaneously hypertensive rat as a model for transmyocardial laser revascularization research [J]. Lasers in Medical ence, 2003, 18(1): 12-18.). Therefore, spontaneously hypertensive rats are a very reliable model for studying hypertension-induced myocardial fibrosis.

[0053] Thirty-two 12-week-old male SHR rats were randomly divided into four groups, with 8 rats in each group:

[0054] Low-dose Chinese medicine group: 120 mg / kg Chuanzhi Tongluo capsule;

[0055] High-dose Chinese medicine group: 240 mg / kg Chuanzhi Tongluo capsule;

[0056] The positive drug group was given captopril 35 mg / kg;

[0057] The SHR blank control group was given an equal volume of distilled water.

[0058] All groups were given the drug by oral gavage for 28 days. At the end of the experiment, the rats were weighed and their blood pressure was measured before being killed.

[0059] Observation and quantitative analysis of myocardial collagen morphology

[0060] A cross section of the left ventricular myocardium was taken approximately 1 mm below the coronary sulcus of the myocardium. The sections were fixed with Boum's solution, embedded in paraffin, and sliced to a thickness of approximately 6 μm. Picric acid Sirius red staining specifically stains collagen, with myocardial cells appearing yellow and collagen appearing red. Measurements were made using an optical microscope, with four fixed fields of view, upper, lower, left, and right, taken from the endothelium and adventitia of each myocardial section. Computer image analysis was used to distinguish collagen from non-collagenous components through grayscale adjustment, and the collagen volume fraction of the myocardial section was measured (CVF = myocardial collagen area / measured field area). Myocardial arterioles are the ratio of the collagen area around the arteriole to the area of the arteriole.

[0061] result

[0062] Effect of the Chinese medicine composition of the present invention on myocardial fibrosis in SHR rats

[0063] The myocardial collagen volume fractions of endocardium, epicardium and arterioles in each group of rats after drug administration are shown in Table 2.

[0064] Compared with the blank control group, the collagen in the ventricle, epicardium and around the myocardial arterioles in each group of the Chinese medicine composition of the present invention was reduced (P < 0.05). The low-dose and high-dose groups of the Chinese medicine composition of the present invention had a significant inhibitory effect on the myocardial fibers of SHR rats.

[0065] Table 2 Effect of the Chinese medicine composition of the present invention on the volume fraction of myocardial collagen in the endocardium, epicardium and arterioles of SHR

[0066]

[0067]

[0068] Compared with the blank group # P<0.05, ## P<0.01.

[0069] Conclusion: This experiment shows that the direct administration of the spontaneous hypertension-induced myocardial fibrosis rat model experiment proves that the Chinese medicine composition of the present invention has the effect of resisting myocardial fibrosis.

[0070] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A use of a Chinese medicine composition prepared from leech, Chuanxiong, astragalus and salvia miltiorrhiza in preparing a drug for preventing and treating myocardial fibrosis, characterized in that: The traditional Chinese medicine composition is prepared from 600 parts by weight of leech, 600 parts by weight of chuanxiong, 400 parts by weight of salvia miltiorrhiza, and 400 parts by weight of astragalus. The myocardial fibrosis is myocardial fibrosis caused by isoproterenol or myocardial fibrosis caused by hypertension.

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