Application of monascus composition in preparation of medicine for treating coronary microcirculation disturbance

Abstract

The invention provides application of a monascus composition in preparation of a medicine for preventing and / or treating coronary microcirculation disturbance. The monascus composition comprises one or more of cortex mori, ramulus mori and folium mori, monascus and radix puerariae. When the composition containing the red yeast rice is applied to preparation of the medicine for preventing and / or controlling coronary microcirculation disturbance, the abnormal microvascular structure can be improved (myocardial microvascular density MVD is increased, edema around microvessels is relieved, the lumen stenosis phenomenon is improved, and the endothelial cell condition is good); 2) microvascular occlusion is improved (pathologic results show that microvascular contents are reduced); (3) the abnormal vasodilatation and contraction function of the microvessels is improved (NO is increased); 4) improving blood lipid metabolism disorder; and (5) the heart function is improved (the left ventricular ejection fraction LVEF and the left ventricular short axis shortening rate LVFS are improved, and the left ventricular pressure maximum rising rate LV + dp / dtmax and the left ventricular pressure maximum falling rate LV-dp / dtmax are increased).

Description

Technical Field

[0001] The present application relates to the field of medicine, and specifically to the use of a red yeast rice composition in the preparation of a medicine for treating coronary microcirculation disorders. Background Art

[0002] The coronary microcirculation consists of anterior arterioles and arterioles. Anterior arterioles (approximately 100-500 mm in diameter) can sense changes in coronary perfusion pressure and / or blood flow, regulating microcirculatory pressure through vasodilation and constriction. Arterioles (<100 mm in diameter) are the site of myocardial metabolic exchange, and their blood flow is primarily influenced by metabolic products.

[0003] Coronary microvascular dysfunction (CMD) refers to a clinical syndrome characterized by myocardial ischemia as the main symptom, caused by functional or structural disorders of the coronary microvessels, leading to insufficient myocardial cell perfusion. It usually occurs in coronary microvessels with a diameter of less than 300 μm and is a common pathogenesis of microcirculatory diseases such as cardiac syndrome X and no-reflow after percutaneous coronary intervention (PCI).

[0004] Studies have found that coronary microcirculatory disorders are closely associated with adverse cardiovascular events. Currently, there is a lack of clear, safe, and effective medications for coronary microcirculatory disorders, both domestically and internationally. In recent years, some researchers have proposed that statins, nicorandil, and calcium antagonists have the potential to improve coronary microcirculatory disorders based on their mechanisms. These studies have led to a series of basic and small-scale clinical studies, but these studies have not yielded satisfactory clinical results. Summary of the Invention

[0005] The present application provides an application of a red yeast rice composition in the preparation of a medicament for treating coronary microcirculation disorders, and provides a new use of the red yeast rice composition.

[0006] The first aspect of the present application provides a use of a red yeast rice composition in preparing a medicament for preventing and / or treating coronary microcirculation disorders, wherein the red yeast rice composition comprises one or more of mulberry bark, mulberry branches and mulberry leaves, red yeast rice and kudzu root.

[0007] The present application includes the application of the above-mentioned red yeast rice composition in the preparation of drugs for preventing and / or treating coronary microcirculatory disorders. Compared with the combination of one or more of mulberry bark, mulberry branch and mulberry leaf and red yeast rice, or the combination of red yeast rice and kudzu root, a synergistic effect is achieved, which can significantly improve microvascular structural abnormalities (increase myocardial microvascular density MVD, reduce edema around microvessels, improve lumen stenosis, and improve endothelial cell condition); 2) improve microvascular obstruction (pathological results show a decrease in microvascular contents); 3) improve microvascular vasomotor function abnormalities (increase NO, reduce ANGⅡ); 4) improve blood lipid metabolism disorders; 5) improve cardiac function (increase left ventricular ejection fraction LVEF and left ventricular short-axis shortening rate LVFS, increase the maximum rate of increase of left ventricular pressure LV+dp / dtmax and the maximum rate of decrease of left ventricular pressure LV-dp / dtmax).

[0008] In any embodiment of the first aspect, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 6-12 parts of white mulberry bark, and 10-15 parts of kudzu root.

[0009] In any embodiment of the first aspect, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 5-10 parts of mulberry leaves, and 10-15 parts of kudzu root.

[0010] In any embodiment of the first aspect, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 9-15 parts of mulberry branches, and 10-15 parts of kudzu roots.

[0011] In any embodiment of the first aspect, the red yeast rice composition comprises, by weight, 6 parts of red yeast rice, 12 parts of mulberry branches, and 12 parts of kudzu roots.

[0012] In any embodiment of the first aspect, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0013] In any embodiment of the first aspect, the pharmaceutically acceptable carrier and / or excipient comprises a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant, a filler, or a disintegrant.

[0014] In any embodiment of the first aspect, the dosage form of the drug comprises an oral administration dosage form.

[0015] In any embodiment of the first aspect, the dosage of red yeast rice is 3-12 g / person / d, the dosage of mulberry bark, mulberry branch or mulberry leaf is 6-12 g / person / d of mulberry bark, 5-10 g / person / d of mulberry leaf or 9-15 g / person / d of mulberry branch, and the dosage of kudzu root is 10-15 g / person / d.

[0016] In any embodiment of the first aspect, the preventing and / or treating of coronary microcirculation disorder comprises one or more of regulating inflammatory response, reducing endothelial cell damage, and increasing myocardial microvessel density. BRIEF DESCRIPTION OF DRAWINGS

[0017] The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiments of the application and together with the description serve to explain the application. In the drawings:

[0018] Figure 1 The results of immunohistochemical detection of CD31 expression (x400) of rats in each experimental group in the present application are shown.

[0019] Figure 2 The results of myocardial tissue HE staining (x200) of rats in each experimental group in the present application are shown.

[0020] Figure 3 The transmission electron microscope images of endothelial cells of rats in each experimental group in the present application are shown. DETAILED DESCRIPTION

[0021] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. The description of the following at least one exemplary embodiment is actually only illustrative, but not as any limitation on the present application and its application or use. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present application.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0023] The present application provides an application of a monascus composition in the preparation of a medicine for preventing and / or treating coronary microcirculation disorder, wherein the monascus composition comprises one or more of mulberry bark, mulberry branch and mulberry leaf, monascus and kudzu root.

[0024] The present application includes the application of the above-mentioned red yeast rice composition in the preparation of a drug for preventing and / or treating coronary microcirculatory disorders. Compared with the combination of one of mulberry bark, mulberry branch and mulberry leaf and red yeast rice, or the combination of red yeast rice and kudzu root, a synergistic effect is achieved, which can significantly improve microvascular structural abnormalities (increase myocardial microvascular density MVD, reduce edema around microvessels, improve lumen stenosis, and improve endothelial cell condition); 2) improve microvascular obstruction (pathological results show a decrease in microvascular contents); 3) improve microvascular vasomotor function abnormalities (increase NO, reduce ANGⅡ); 4) improve blood lipid metabolism disorders; 5) improve cardiac function (increase left ventricular ejection fraction LVEF and left ventricular short-axis shortening rate LVFS, increase the maximum rate of increase of left ventricular pressure LV+dp / dtmax and the maximum rate of decrease of left ventricular pressure LV-dp / dtmax).

[0025] The red yeast rice of the present application can be obtained by directly crushing the red yeast rice medicinal material, preferably through a 40-60 mesh sieve or a 20-80 mesh sieve.

[0026] The mulberry bark, mulberry branch or mulberry leaf and kudzu root of the present application can be extracted by conventional extraction methods. In some embodiments, the respective extraction methods are exemplified, but the following examples should not be regarded as limiting the scope of protection of the present application.

[0027] Add 5-10 times its weight of water to the bark of Morus alba, soak for 1-3 hours, decoct 1-3 times, each time for 1-2 hours, filter, combine the filtrate and concentrate it into an extract, put the extract into a vacuum drying oven (-0.05--0.09Mpa pressure, 60℃-80℃ drying), grind it and pass it through a 40-mesh sieve to obtain the bark of Morus alba extract powder.

[0028] Add 5-10 times the weight of water to mulberry leaves, boil 1-3 times, each time for 1-2 hours, filter, combine the filtrate and concentrate it into an extract, put the extract into a vacuum drying oven (-0.05--0.09Mpa pressure, 60℃-80℃ drying), crush it and pass it through a 40-mesh sieve to obtain mulberry leaf extract powder.

[0029] Add 5-10 times the weight of water to the mulberry branches, boil for 1-3 times, each time for 1-2 hours, filter, combine the filtrate and concentrate it into an extract, put the extract into a vacuum drying oven (-0.05--0.09Mpa pressure, 60℃-80℃ drying), crush it and pass it through a 40-mesh sieve to obtain mulberry branch extract powder.

[0030] Add 5-10 times its weight of water to the kudzu root slices, soak for 1-3 hours, boil the water and then boil for 1-3 times, each time for 1-2 hours, filter, combine the filtrate and concentrate to obtain an extract, put the extract into a reduced pressure drying oven (-0.05--0.09Mpa pressure, 60℃-80℃ drying), crush and pass through a 40-mesh sieve to obtain kudzu root extract powder, which is used in this application.

[0031] In some embodiments, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 6-12 parts of white mulberry bark, and 10-15 parts of kudzu root.

[0032] In some embodiments, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 5-10 parts of mulberry leaves, and 10-15 parts of kudzu root.

[0033] In some embodiments, the red yeast rice composition comprises, by weight, 3-12 parts of red yeast rice, 9-15 parts of mulberry branches, and 10-15 parts of kudzu roots.

[0034] In some embodiments, the red yeast rice composition comprises 6 parts of red yeast rice, 12 parts of mulberry branches and 12 parts of kudzu roots, in parts by weight. In some embodiments, the drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0035] In some embodiments, the pharmaceutically acceptable carrier and / or excipient includes a diluent, a binder, a surfactant, a wetting agent, an absorbent carrier, a lubricant, a filler, or a disintegrant.

[0036] In some embodiments, the dosage form of the drug comprises an oral dosage form.

[0037] In some embodiments, the dosage of red yeast rice is 3-12 g / person / d, the dosage of mulberry bark, mulberry branch or mulberry leaf is: mulberry bark 6-12 g / person / d, mulberry leaf 5-10 g / person / d or mulberry branch 9-15 g / person / d, and the dosage of kudzu root is 10-15 g / person / d.

[0038] In some embodiments, preventing and / or treating coronary microcirculatory disorders includes one or more of regulating inflammatory response, reducing endothelial cell damage, and increasing myocardial microvascular density.

[0039] The beneficial effects of the present application will be further illustrated below with reference to examples and comparative examples, but these should not be construed as limiting the scope of protection of the present application.

[0040] [Experimental reagents]

[0041] Sodium laurate solution (lot number: 01591945), Sigma Co., Ltd., USA. Penicillin (lot number: 080451253), Zhongjing Biotechnology Co., Ltd.; isoflurane (lot number: G45992), Beijing Yizejia Technology Co., Ltd.; total cholesterol (TC) kit (lot number: 20240809), triglyceride (TG) kit (20240809), low-density lipoprotein (LDL) kit (lot number: 20240813), high-density lipoprotein (HDL) kit (lot number: 20240809), endothelin-1 (ET-1) kit (lot number: 20240830), Nitric oxide (NO) detection kit by nitric acid reduction method (Batch No.: 20240826) was purchased from Nanjing Jiancheng Biological Research Institute; rat vascular cell adhesion molecule 1 (VCAM-1) enzyme-linked immunosorbent assay kit (Batch No.: 20250203.60718R) and rat angiotensin II (ANG-II) enzyme-linked immunosorbent assay kit (Batch No.: 20250302.60001R) were purchased from Beijing Rigbo Biotechnology Co., Ltd.; rat tumor necrosis factor α (TNF-α) kit (Batch No. : 202411), rat thromboxane B2 (TXB2) kit (batch number: 202411), purchased from Wuhan Yilai Ruite Biotechnology Co., Ltd.; hematoxylin-eosin (HE) staining kit (batch number: G1120), all purchased from Beijing Solarbio Company; BCA protein quantification kit (Yazyme Biotechnology Co., Ltd., batch number: ZJ102); B lymphocytoma-2 (Bcl-2) polyclonal antibody (batch number: 26593-1-AP), Bcl-2 associated protein X (BAX) polyclonal antibody (batch number: :50599-2-AP), caspase-3 polyclonal antibody (lot number: 25128-1-AP), all purchased from Wuhan Proteintech Biological Co., Ltd.; β-actin antibody (lot number: bs-10900R); vascular endothelial growth factor (VEGF) antibody (lot number: bs-0279R); rat anti-CD31 antibody (lot number: bs-0195R), all purchased from Beijing Bioss Biological Co., Ltd.; goat anti-rabbit IgG H&L (lot number: ab150077), all purchased from Abcam, USA. Red yeast rice powder (lot number: Y202310190) was purchased from Peking University Weixin Biological Co., Ltd.

[0042] Pueraria root extraction: Take 3kg of Pueraria root slices, add 8 times the amount (24L) of water, soak for 2h, boil the water and decoct twice, each time for 1.5h, combine the filtrates, concentrate to obtain the extract, and dehydrate in a vacuum drying oven at -0.07MPa pressure and 70℃. Grind and pass through a 40-mesh sieve.

[0043] Mulberry branch extraction: Take 3kg of mulberry branches and add 8 times the amount (24L) of water. Soak for 2h and then boil twice, each time for 1-2 hours. Filter, combine the filtrate and concentrate it into an extract. Put the extract into a vacuum drying oven (-0.05--0.09Mpa pressure, 60℃-80℃ drying), crush it and pass it through a 40-mesh sieve to obtain mulberry branch extract powder.

[0044] [Experimental instruments]

[0045] Small animal anesthesia machine (Shenghe Aizhong Medical Technology Co., Ltd.); BL-420I animal ventilator (Chengdu Taimeng Software Co., Ltd.); PowerPac TM HC high current electrophoresis instrument, Mini- Tetra type vertical protein electrophoresis tank, Mini-Trans- The Module-type slot transfer system was purchased from Shanghai Bio-Rad Biomedical Products Co., Ltd.; the MuttiSkan Mk3 microplate reader was purchased from Thermo Fisher Scientific, USA; the HT7800 transmission electron microscope was purchased from Hitachi, Ltd., Japan; and the Leica UC7 ultrathin microtome was purchased from Leica Microsystems, Germany.

[0046] [Experimental Animals]:

[0047] SPF-grade Sprague-Dawley rats, half male and half female, weighing 120 g to 150 g, were obtained from Sibeifu (Beijing) Biotechnology Co., Ltd. (Production License: SCXK (Beijing) 2019-0010, Animal Qualification Certificate No.: 110324241100966447). The rats were maintained at a constant temperature of (25 ± 2)°C and a relative humidity of (60 ± 5)%.

[0048] [Experimental Grouping]

[0049] The control group was fed with normal maintenance feed, and the other groups were fed with high-fat feed after 3 days of adaptive feeding; according to the different modeling and drug administration methods, they were divided into: high-fat feeding group, positive drug nicorandil group (positive drug group), red yeast rice + kudzu root group, red yeast rice + mulberry branch group, red yeast rice + kudzu root + mulberry branch group and model group.

[0050] The clinical daily dosage for humans is 6g of red yeast rice and 12g of kudzu root in the red yeast rice + mulberry branch group; 6g of red yeast rice and 12g of mulberry branch in the red yeast rice + kudzu root + mulberry branch group; and 6g of red yeast rice, 12g of kudzu root, and 12g of mulberry branch in the red yeast rice + kudzu root + mulberry branch group. Assuming the proportions of each component remain constant within each group, the equivalent daily dosage for rats is 1.8g (crude drug) / kg for the red yeast rice + kudzu root group, 1.8g (crude drug) / kg for the red yeast rice + mulberry branch group, and 3.0g (crude drug) / kg for the red yeast rice + kudzu root + mulberry branch group. The clinical dosage of Nicorandil tablets is 15mg / human / day, which translates to a rat dosage of 1.5mg / kg.

[0051] [Animal modeling method]

[0052] The control group was fed a normal diet and water, and after four weeks, blood was collected from the inner canthus for lipid testing. After one week of adaptive feeding, healthy SD rats were fed a high-fat diet for four weeks to induce hyperlipidemia. After four weeks, blood was collected from the inner canthus for lipid testing. After confirming dyslipidemia, the rats were divided into six groups based on cholesterol (TC) values: a high-fat diet group, a model group, a positive drug group (nicorandil), a red yeast rice + kudzu root group, a red yeast rice + mulberry branch group, and a red yeast rice + kudzu root + mulberry branch group. The mean TC values ​​of the rats in each group were similar, and preventive medication was then administered for two weeks.

[0053] Two weeks after the preventive treatment, rats in all groups except the control group were subjected to a rat model of coronary microcirculatory dysfunction. The rats were anesthetized with 4% isoflurane, intubated, and connected to a small animal ventilator and a gas anesthesia machine for maintenance. The left thorax was prepared, and the skin was incised obliquely along the 2nd to 4th ribs to the left of the sternum. The muscular layer was bluntly dissected, and the 3rd and 4th intercostal spaces were stretched to fully expose the heart and aorta. The aortic arch beneath the thymus was carefully dissected with forceps, and a cotton thread was passed under the arch. The heart was carefully retrieved from the thorax and gently lifted. The aorta was clamped with a vascular clamp, and a 4 mg / kg sodium laurate solution was simultaneously injected rapidly into the apex of the heart using an insulin syringe. After maintaining the clamp for 20 seconds, the clamp was released, the heart was intubated, air was expelled from the thorax, and the wound was sutured layer by layer. The sodium laurate solution was kept warm in a 37°C water bath to prevent coagulation of the sodium laurate solution. After surgery, 40,000 units of penicillin were injected intramuscularly to prevent wound infection.

[0054] The high-fat feeding group also received the above treatment, but an equal amount of normal saline was used instead of sodium laurate solution. The rats in the high-fat feeding group continued to be fed with high fat after the model was established, and the samples were collected after 3 weeks.

[0055] The control group continued to be fed with normal food and water, and samples were collected after 3 weeks.

[0056] The rats in the model group continued to be fed with high-fat diet for 3 weeks before samples were collected.

[0057] After the rat models were established, the rats in each drug-treated group continued to be fed with high-fat diet and given drugs for 3 weeks before the samples were collected.

[0058] [test]

[0059] Echocardiography

[0060] Three weeks after surgery, rats were anesthetized with an intraperitoneal injection of 1.5% sodium pentobarbital solution at a dose of 0.3 mL / 100 g on day 21. The skin was prepared from the right midclavicular line to the left midaxillary line. Ultrasound observation was performed on the parasternal short-axis section with a high-frequency probe in M-mode. The average left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) of three cardiac cycles were calculated.

[0061] Hemodynamic testing

[0062] On the 21st day after postoperative medication, rats were anesthetized with an intraperitoneal injection of 1.5% sodium pentobarbital solution at a dose of 0.3 mL / 100 g. The rats were fixed on the operating board, the skin and muscles of the neck were cut layer by layer, the right common carotid artery was freed, the distal end was ligated, and the proximal end was clamped with an arterial clamp. After the arterial retention needle was inserted and the arterial clamp was released, the tail end of the arterial retention needle was connected to the BL-420I system to record the heart rate, the maximum rate of increase of left ventricular pressure (LV+dp / dtmax), and the maximum rate of decrease of left ventricular pressure (LV-dp / dtmax).

[0063] Detection of changes in rat serum related indicators

[0064] After hemodynamic testing, blood was collected from the abdominal aorta of each group of rats. The blood samples were kept at room temperature for 2 hours and centrifuged at 3500 rpm for 10 minutes to obtain serum. Serum nitric oxide (NO) levels were measured by biochemical analysis, and serum endothelin (ET-1), vascular cell adhesion molecule-1 (VCAM-1), and angiotensin II (ANGⅡ) levels were measured by ELISA.

[0065] Myocardial ultrastructural observation

[0066] 21 days after surgery, the rats were anesthetized and the hearts were removed, washed with pre-cooled saline, and dried with filter paper. The heart weight was weighed and the cardiac organ index was calculated (heart weight / body weight*100). 3 The left ventricular tissues of different sizes were fixed with 3.5% glutaraldehyde, embedded with conventional electron microscopy technology, and ultrathin sections (thickness 50nm) were cut. After double staining with uranyl acetate and lead citrate, the ultrastructural changes of rat myocardial microvessels were observed under transmission electron microscopy.

[0067] Myocardial microvascular density detection

[0068] 21 days after surgery, heart tissue was obtained and fixed in 10% neutral malonic acid, embedded in paraffin, sectioned, and CD31 antibody was added and incubated at 4°C overnight. Secondary antibody was added and incubated at room temperature for 1 hour. DAB was used for color development, hematoxylin was used for counterstaining, and the slides were mounted with neutral gum. Under a microscope, vascular endothelial cells were observed to be brown-yellow stained. Image J software was used for analysis, and the number of vascular endothelial cells per unit area was calculated, which was the microvessel density (MVD).

[0069] Western blotting to detect related protein expression

[0070] Cardiac tissue from each group was removed from a -80°C freezer and placed on ice. RIPA lysis buffer and protease and phosphatase inhibitors were added, followed by mechanical homogenization and centrifugation. The protein concentration of the supernatant was determined using the BCA protein assay, and the protein concentration of each sample was adjusted. Total myocardial tissue protein (10 μg) was separated by SDS-PAGE and then electrotransferred onto a PVDF membrane. The PVDF membrane containing proteins was blocked with skim milk powder for 1.5 hours and then incubated with primary antibodies overnight at 4°C. The following antibodies were used: VEGF (1:2000), Bcl-2 (1:2000), BAX (1:2000), caspase-3, GADPH (1:5000), and β-actin (1:5000). The membrane was washed three times with TBST and incubated with goat anti-rabbit IgG HRP (1:5000) or goat anti-mouse IgG HRP (1:5000) for 1 hour at room temperature. Protein band signals were detected using an Amersham ECL system. Image J software was used to quantitatively analyze protein bands.

[0071] [Statistics and Analysis]

[0072] Data are average Data are expressed as mean ± standard deviation (SD). Statistical comparisons among groups were performed using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test. Statistical significance was defined as P < 0.05 or P < 0.01.

[0073] [result]

[0074] (1) Effects of red yeast rice composition on body weight and cardiac organ index in rats with coronary microcirculatory disorder induced by sodium laurate injection and high-fat diet

[0075] Table 1 Test results of rat body weight and organ index

[0076]

[0077] Note: Compared with the control group, △△ P<0.01; compared with the model group, ▲ P<0.05, ▲▲ P<0.01.

[0078] Compared with the control group, the body weight of rats with coronary microcirculation disorder induced by sodium laurate injection plus high-fat diet in the model group decreased to a certain extent; compared with the model group, there was no significant difference in body weight among the red yeast rice + mulberry branch group, red yeast rice + kudzu root group, and red yeast rice + kudzu root + mulberry branch group, among which the body weight of the red yeast rice + kudzu root + mulberry branch group showed a trend of increasing.

[0079] Compared with the control group, the cardiac organ index of rats in the model group with coronary microcirculatory impairment induced by sodium laurate injection and high-fat diet was significantly decreased, indicating the presence of heart disease. Compared with the model group, the cardiac organ index of rats in the red yeast rice + kudzu root group and the red yeast rice + kudzu root + mulberry branch group significantly recovered.

[0080] (2) Effects of red yeast rice composition on echocardiography in rats with coronary microcirculatory disorders induced by sodium laurate injection and high-fat diet

[0081] Table 2 LVEF and LVFS test results of rats

[0082]

[0083]

[0084] Note: Compared with the control group, △ P<0.05, △△ P<0.01; compared with the model group, ▲ P<0.05, ▲▲ P<0.01.

[0085] Compared with the control group, the cardiac function-related indicators of the rat model with coronary microcirculation disorder induced by sodium laurate injection and high-fat diet in the model group, including left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), were significantly decreased, indicating that the rats in the model group had obvious cardiac contractile function damage.

[0086] Compared with the model group, the administration of red yeast rice + kudzu root and red yeast rice + kudzu root + mulberry twig can significantly increase the LVEF and LVFS levels, and significantly and effectively improve the cardiac contractile function of the model animals; the administration of red yeast rice + mulberry twig has a trend of increasing LVEF and LVFS, but no significant difference was observed.

[0087] (3) Effects of red yeast rice composition on hemodynamics in rats with coronary microcirculatory disorders induced by sodium laurate injection and high-fat diet

[0088] Table 3 Hemodynamic test results of rats

[0089]

[0090] Note: Compared with the control group, △ P<0.05, △△ P<0.01; compared with the model group, ▲ P<0.05, ▲▲ P<0.01.

[0091] Compared with the control group, the heart rate of the rat model with coronary microcirculation disorder induced by sodium laurate injection plus high-fat diet in the model group was significantly decreased, and the absolute values ​​of the maximum rate of increase of left ventricular pressure (LV+dp / dtmax) and the maximum rate of decrease of left ventricular pressure (LV-dp / dtmax) were significantly reduced, indicating that the animals had obvious cardiac systolic and diastolic function disorders.

[0092] Compared with the model group of rats with coronary microcirculatory dysfunction induced by sodium laurate injection and high-fat diet, administration of red yeast rice combined with kudzu root and red yeast rice combined with kudzu root combined with mulberry twigs significantly improved LV+dp / dtmax and LV-dp / dtmax in rats, significantly and effectively improving cardiac function in rats with coronary microcirculatory dysfunction. Administration of red yeast rice combined with mulberry twigs did not significantly improve hemodynamics.

[0093] (4) Effects of red yeast rice composition on endothelial function markers in rats with coronary microcirculatory disorders induced by sodium laurate injection and high-fat diet

[0094] Table 4 Test results of rat vascular endothelial function markers

[0095]

[0096] Note: Compared with the control group, △ P<0.05, △△ P<0.01; compared with the model group, ▲ P<0.05, ▲▲ P<0.01.

[0097] Compared with the control group, the vascular endothelial function of the rat model of coronary microcirculation disorder induced by sodium laurate injection plus high-fat diet in the model group was significantly damaged, as manifested by a significant decrease in blood NO and a significant increase in ET-1, VCAM-1, and ANGⅡ levels, indicating abnormal vascular function.

[0098] Compared with the model group, the administration of red yeast rice + kudzu root, red yeast rice + kudzu root + mulberry branch can significantly increase the NO level and reduce the ET-1 and ANGII levels; the administration of red yeast rice + kudzu root, red yeast rice + mulberry branch, red yeast rice + kudzu root + mulberry branch can significantly reduce the VCAM-1 level; the administration of red yeast rice + kudzu root, red yeast rice + kudzu root + mulberry branch can significantly reduce the ANGII level.

[0099] (5) Effect of red yeast rice composition on myocardial microvascular density in rats with coronary microcirculatory disorder induced by sodium laurate injection and high-fat diet

[0100] Table 5 Rat microvessel density test results

[0101]

[0102]

[0103] Note: Compared with the control group, △ P<0.05, △△ P<0.01; compared with the model group, ▲ P<0.05, ▲▲ P<0.01.

[0104] Compared with the control group, the myocardial microvascular density (MVD) of rats with coronary microcirculation disorder induced by sodium laurate injection plus high-fat diet in the model group was significantly decreased, and microcirculation disorder was obvious.

[0105] Compared with the model group, administration of red yeast rice + kudzu root and red yeast rice + kudzu root + mulberry branch can significantly and effectively increase the microvascular density of rats.

[0106] [Immunohistochemical detection of CD31 expression]

[0107] The results are recorded in Figure 1 middle. Figure 1 It can be seen that compared with the model group, the microvascular density of each compound drug group increased significantly (P<0.05).

[0108] [Effects on myocardial tissue morphology in rats with microvascular injury]

[0109] Myocardial tissue morphology in rats with microvascular injury Figure 2 .

[0110] In the control group, the myocardial cells were tightly arranged, the cell structure was intact, the macrovascular and microvascular structures were intact without obvious lesions, and there was no inflammatory infiltration.

[0111] The rats in the model group showed obvious myocardial cell dissolution and necrosis, large-area inflammatory infiltration, obvious microvascular structural damage, perivascular edema and a large amount of contents in the blood vessels.

[0112] Compared with the model group, the myocardial inflammatory infiltration in the red yeast rice + kudzu root group was improved, and the microvascular structure became more intact. The red yeast rice + mulberry branch group showed significant improvement in myocardial inflammatory infiltration, with mild damage and edema in the microvascular wall. The red yeast rice + kudzu root + mulberry branch group showed significant improvements in myocardial arrangement and cellular integrity, as well as significant improvement in inflammatory infiltration, with relatively intact microvascular walls and mild edema. This improvement in myocardial tissue morphology was synergistic with the other two treatment groups.

[0113] [Effects on rat myocardial ultrastructure]

[0114] Ultrastructural changes of rat myocardium Figure 3 .

[0115] In the control group, the tight junctions of myocardial endothelial cells were intact, the basement membrane structure was intact, there was no stenosis or deformation of the lumen, no edema outside the microvessels, and the myocardial mitochondrial structure around the blood vessels was clear and intact.

[0116] In the model group, the tight junctions of myocardial endothelial cells were significantly damaged, the basement membrane structure was discontinuous, vacuoles were present in the vascular wall, the lumen was narrowed and deformed, and the mitochondrial cristae structure around the blood vessels was unclear.

[0117] The loss of tight junctions of endothelial cells in rats in the red yeast rice + kudzu root group, red yeast rice + mulberry branch group, and red yeast rice + kudzu root + mulberry branch group was improved, the luminal stenosis was improved, the perivascular edema was reduced, and a small number of vacuolar areas were present on the vascular wall, indicating that the condition of microvascular endothelial cells was improved; the structure of myocardial mitochondria around the blood vessels was clearer.

[0118] Data analysis of the synergistic effects of red yeast rice, kudzu root, and mulberry branches:

[0119] The coefficient of drug interaction (CDI) was calculated as AB / (A×B), where AB is the ratio of the experimental results of the red yeast rice + kudzu root + mulberry branch group to the model group, and A and B are the ratios of the experimental results of the red yeast rice + mulberry branch group and the red yeast rice + kudzu root group to the model group, respectively.

[0120] When CDI < 1, it indicates the presence of synergistic effect. The calculation results are recorded in Table 6. Table 6

[0121]

[0122] The results showed that compared with the two-component compound group, the three-component compound group showed a synergistic effect in organ index, LVEF, LVFS, LV+dp / dtmax, LV-dp / dtmax, NO and MVD indicators (CDI < 1), suggesting that the compound composed of red yeast rice, kudzu root and mulberry twig can cooperate with each other to improve the efficacy of myocardial tissue, cardiac function, vasodilation function and angiogenesis in the model group rats.

[0123] in conclusion:

[0124] The core pathologies of coronary microcirculatory disorders include abnormal microvascular structure, microvascular obstruction and abnormal microvascular vasodilation and contraction function. The study successfully simulated clinical coronary microcirculatory disorders by injecting sodium laurate and feeding rats with high-fat diet, and on this basis, the function of red yeast rice composition was studied.

[0125] The results showed that the red yeast rice combination of red yeast rice + kudzu root + mulberry branch showed a clear improvement effect on coronary microcirculation disorders, including 1) improvement of microvascular structural abnormalities (increased myocardial microvascular density MVD, reduced peri-microvascular edema, improved lumen stenosis, and good endothelial cell condition); 2) improvement of microvascular obstruction (pathological results showed a decrease in microvascular contents); 3) improvement of microvascular vasomotor function abnormalities (increased NO and reduced ANGⅡ); 4) improvement of lipid metabolism disorders; 5) improvement of cardiac function (increased left ventricular ejection fraction LVEF and left ventricular short-axis shortening rate LVFS, increased the maximum rate of increase of left ventricular pressure LV+dp / dtmax and the maximum rate of decrease of left ventricular pressure LV-dp / dtmax). 6) The three components of red yeast rice, kudzu root, and mulberry twig showed synergistic effects in organ index, LVEF, LVFS, LV+dp / dtmax, LV-dp / dtmax, NO and MVD indicators (CDI < 1), indicating that the compound composed of red yeast rice, kudzu root, and mulberry twig can cooperate with each other to improve the therapeutic effect on myocardial tissue, cardiac function, vasodilation function and angiogenesis in the model group rats.

[0126] The above describes exemplary embodiments of the present invention. However, the scope of protection of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A use of a red yeast rice composition in preparing a medicament for preventing and / or treating coronary microcirculation disorders, the red yeast rice composition comprising one or more of mulberry bark, mulberry branch and mulberry leaf, red yeast rice and kudzu root.

2. The use according to claim 1, wherein In parts by weight, the red yeast rice composition comprises 3-12 parts of red yeast rice, 6-12 parts of white mulberry bark and 10-15 parts of kudzu root; or In parts by weight, the red yeast rice composition comprises 3-12 parts of red yeast rice, 5-10 parts of mulberry leaves and 10-15 parts of kudzu root; or In parts by weight, the red yeast rice composition comprises 3-12 parts of red yeast rice, 9-15 parts of mulberry branches and 10-15 parts of kudzu roots.

3. The use according to claim 1, wherein: In parts by weight, the red yeast rice composition comprises 6 parts of red yeast rice, 12 parts of mulberry branches and 12 parts of kudzu roots.

4. The use according to any one of claims 1 to 3, wherein The medicine further includes pharmaceutically acceptable carriers and / or excipients.

5. The use according to claim 4, wherein: The pharmaceutically acceptable carriers and / or excipients include diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, or disintegrants.

6. The use according to any one of claims 1 to 5, wherein The dosage form of the drug includes an oral dosage form.

7. The use according to any one of claims 1 to 6, wherein The dosage of the red yeast rice is 3-12 g / person / d, the dosage of the mulberry bark, mulberry branch or mulberry leaf is 6-12 g / person / d of the mulberry bark, 5-10 g / person / d of the mulberry leaf or 9-15 g / person / d of the mulberry branch, and the dosage of the kudzu root is 10-15 g / person / d.

8. The use according to any one of claims 1 to 7, wherein: The prevention and / or treatment of coronary microcirculation disorders includes one or more of regulating inflammatory response, alleviating endothelial cell damage, and increasing myocardial microvascular density.

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