Use of a monascus composition in the preparation of a medicament for treating coronary microcirculation disorder

Abstract

The application provides application of a monascus composition in 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 pueraria. When the monascus composition is applied in preparation of the medicine for preventing and / or treating coronary microcirculation disorder, the following effects can be achieved: 1) improving microvascular structure abnormality (increasing myocardial microvascular density MVD, reducing perivascular edema of microvessels, improving lumen stenosis phenomenon, and good endothelial cell condition); 2) improving microvascular obstruction (pathological results show that the content of microvessels is reduced); 3) improving abnormal microvascular vasomotor function (increasing NO); 4) improving blood lipid metabolism disorder; and 5) improving heart function (increasing left ventricular ejection fraction LVEF and left ventricular short axis shortening rate LVFS, increasing left ventricular internal pressure maximum rising rate LV+dp / dtmax and left ventricular internal pressure maximum falling rate LV‑dp / dtmax).

Description

Technical Field

[0001] This application relates to the pharmaceutical field, specifically to the use of a red yeast rice composition in the preparation of a medicament for treating coronary microcirculation disorders. Background Technology

[0002] The coronary microcirculation includes 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 vasoconstriction. Arterioles (<100 mm in diameter) are the site of myocardial metabolic exchange, and their blood flow is mainly affected by metabolic products.

[0003] Coronary microvascular dysfunction (CMD) refers to a clinical syndrome characterized by myocardial ischemia due to insufficient myocardial perfusion caused by functional or structural disorders of coronary microvessels. 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 a close link between coronary microcirculatory disturbances and adverse cardiovascular events. Currently, there is a lack of clearly safe and effective drugs for intervening in coronary microcirculatory disturbances both domestically and internationally. In recent years, some scholars have proposed that statins, nicorandil, and calcium channel blockers have the potential to improve coronary microcirculatory disturbances by focusing on their mechanisms, and have conducted a series of basic and small-scale clinical studies, but have not achieved ideal clinical results. Summary of the Invention

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

[0006] The first aspect of this application provides the use of a red yeast rice composition in the preparation of a medicament for the prevention and / or treatment of coronary microcirculation disorders, the red yeast rice composition comprising one or more of mulberry bark, mulberry twigs and mulberry leaves, red yeast rice and kudzu root.

[0007] When the above-mentioned red yeast rice composition is used in the preparation of a drug for preventing and / or improving coronary microcirculatory disorders, this application achieves a synergistic effect relative to one or more of mulberry bark, mulberry twigs and mulberry leaves and red yeast rice or a combination of red yeast rice and kudzu root. It can significantly improve abnormal microvascular structure (increase myocardial microvascular density MVD, reduce perivascular edema, improve luminal stenosis, and improve endothelial cell condition); 2) improve microvascular obstruction (pathological results show a reduction in microvascular contents); 3) improve abnormal microvascular vasomotor function (increase NO, decrease ANGII); 4) improve lipid metabolism disorders; 5) improve cardiac function (increase left ventricular ejection fraction LVEF and left ventricular fractional shortening LVFS, increase the maximum rate of rise of left ventricular intraventricular pressure LV+dp / dtmax and the maximum rate of fall of left ventricular intraventricular pressure LV-dp / dtmax).

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

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

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

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

[0012] In any embodiment of the first aspect, the medicament further includes a pharmaceutically acceptable carrier and / or excipients.

[0013] In any embodiment of the first aspect, pharmaceutically acceptable carriers and / or excipients include diluents, binders, surfactants, humectants, adsorbents, lubricants, fillers, or disintegrants.

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

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

[0016] In any embodiment of the first aspect, prevention and / or treatment of coronary microcirculation disorders includes one or more of the following: modulating inflammatory responses, reducing endothelial cell damage, and increasing myocardial microvascular density. Attached Figure Description

[0017] The accompanying drawings, which are included to provide a further understanding of the invention and form part of this invention, illustrate exemplary embodiments of the invention and are used to explain the invention, but do not constitute an undue limitation of the invention. In the drawings:

[0018] Figure 1 The results of immunohistochemical detection of CD31 expression (×400) in rats of each experimental group in this application are shown.

[0019] Figure 2 The results of HE staining (×200) of myocardial tissue from rats in each experimental group of this application are shown.

[0020] Figure 3 Transmission electron micrographs of rat endothelial cells from each experimental group in this application are shown. Detailed Implementation

[0021] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[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 this specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0023] This application provides the use of a red yeast rice composition in the preparation of a medicament for the prevention and / or treatment of coronary microcirculation disorders. The red yeast rice composition includes one or more of mulberry bark, mulberry twigs and mulberry leaves, red yeast rice, and kudzu root.

[0024] When the above-mentioned red yeast rice composition is used in the preparation of a drug for preventing and / or improving coronary microcirculatory disorders, this application achieves a synergistic effect relative to the combination of red yeast rice with one of mulberry bark, mulberry twig and mulberry leaf, or the combination of red yeast rice and kudzu root. It can significantly improve abnormal microvascular structure (increase myocardial microvascular density MVD, reduce perivascular edema, improve luminal stenosis, and improve endothelial cell condition); 2) improve microvascular obstruction (pathological results show a reduction in microvascular contents); 3) improve abnormal microvascular vasomotor function (increase NO, decrease ANGII); 4) improve lipid metabolism disorders; 5) improve cardiac function (increase left ventricular ejection fraction LVEF and left ventricular fractional shortening LVFS, increase the maximum rate of rise of left ventricular pressure LV+dp / dtmax and the maximum rate of fall of left ventricular pressure LV-dp / dtmax).

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

[0026] The mulberry bark, twigs or leaves and kudzu root of this application can be extracted using conventional extraction methods. In some embodiments, the respective extraction methods are illustrated by way of example, but the following examples should not be regarded as limiting the scope of protection of this application.

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

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

[0029] Add 5-10 times the weight of water to mulberry branches, decoct 1-3 times, 1-2 hours each time, filter, combine the filtrates and concentrate into an extract, put the extract into a vacuum drying oven (-0.05--0.09 MPa pressure, 60℃-80℃ drying), pulverize and pass 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 decoct 1-3 times, 1-2 hours each time, filter, combine the filtrates and concentrate to obtain an extract, put the extract into a vacuum drying oven (-0.05--0.09 MPa pressure, 60℃-80℃ drying), pulverize 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 red yeast rice, 6-12 parts mulberry bark, and 10-15 parts kudzu root.

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

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

[0034] In some embodiments, the red yeast rice composition comprises, by weight, 6 parts red yeast rice, 12 parts mulberry twigs, and 12 parts kudzu root. In some embodiments, the drug further comprises a pharmaceutically acceptable carrier and / or excipients.

[0035] In some embodiments, pharmaceutically acceptable carriers and / or excipients include diluents, binders, surfactants, humectants, adsorbents, lubricants, fillers, or disintegrants.

[0036] In some implementations, the dosage form of the drug includes an oral dosage form.

[0037] In some implementations, the dosage of red yeast rice is 3-12g / person / day, and the dosage of mulberry bark, mulberry twigs or mulberry leaves is: 6-12g / person / day for mulberry bark, 5-10g / person / day for mulberry leaves or 9-15g / person / day for mulberry twigs, and 10-15g / person / day for kudzu root.

[0038] In some implementations, prevention and / or treatment of coronary microcirculatory disorders include one or more of the following: modulating the inflammatory response, reducing endothelial cell damage, and increasing myocardial microvascular density.

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

[0040] [Experimental Reagents]

[0041] Sodium lauryl solution (batch number: 01591945), Sigma-Aldrich, Inc., USA. Penicillin (batch number: 080451253), Zhongjing Biotechnology Co., Ltd.; Isoflurane (batch number: G45992), Beijing Yizejia Technology Co., Ltd.; Total cholesterol (TC) kit (batch number: 20240809), triglyceride (TG) kit (20240809), low-density lipoprotein (LDL) kit (batch number: 20240813), high-density lipoprotein (HDL) kit (batch number: 20240809), endothelin-1 (ET-1) kit (batch number: 20240830). Nitric oxide (NO) detection kit by nitrate reduction method (batch number: 20240826), all kits were purchased from Nanjing Jiancheng Biological Institute; rat vascular cell adhesion molecule-1 (VCAM-1) enzyme-linked immunosorbent assay kit (batch number: 20250203.60718R), rat angiotensin II (ANG-II) enzyme-linked immunosorbent assay kit (batch number: 20250302.60001R), all kits were purchased from Beijing Ruigebo Biotechnology Co., Ltd.; rat tumor necrosis factor α (TNF-α) kit (batch number: ... Rat thromboxane B2 (TXB2) kit (batch number: 202411), purchased from Wuhan Yilairuit Biotechnology Co., Ltd.; Hematoxylin-eosin (HE) staining kit (batch number: G1120), both purchased from Beijing Solarbio Co., Ltd.; BCA protein quantification kit (Yamei Biotechnology Co., Ltd., batch number: ZJ102); B lymphoma-2 (Bcl-2) polyclonal antibody (batch number: 26593-1-AP), Bcl-2 related protein X (BAX) polyclonal antibody (batch number: 202411), purchased from Wuhan Yilairuit Biotechnology Co., Ltd.; Hematoxylin-eosin (HE) staining kit (batch number: G1120), both purchased from Beijing Solarbio Co., Ltd.; BCA protein quantification kit (Yamei Biotechnology Co., Ltd., batch number: ZJ102); B lymphoma-2 (Bcl-2) polyclonal antibody (batch number: 26593-1-AP), purchased from Beijing Solarbio Co., Ltd.; BCA protein quantification kit (Yamei Biotechnology Co., Ltd., batch number: ZJ102); B lymphoma-2 (Bcl-2) polyclonal antibody (batch number: 202411), purchased from Wuhan Yilairuit Biotechnology Co., Ltd.; Hematoxylin-eosin (HE) staining kit (batch number: G1120), both purchased from Beijing Solarbio Co., Ltd.; BCA protein quantification kit (Yamei Biotechnology Co., Ltd., batch number: ZJ102); B lymphoma-2 (Bcl-2) polyclonal antibody (batch number: 202411), purchased from Wuhan Yilairuit Biotechnology Co., Ltd.; Bcl-2 related protein X (BAX) polyclonal antibody (batch number: 202411), purchased from Wuhan Yilairuit Biotechnology Co., Ltd.; The following antibodies were purchased from Proteintech Biotechnology Co., Ltd.: 50599-2-AP, cysteine-aspartic protease (Caspase-3) polyclonal antibody (batch number: 25128-1-AP); β-actin antibody (batch number: bs-10900R); vascular endothelial growth factor (VEGF) antibody (batch number: bs-0279R); rat anti-CD31 antibody (batch number: bs-0195R); and goat anti-rabbit IgG H&L (batch number: ab150077). All of these antibodies were purchased from Abcam Biotechnology Co., Ltd. (USA). Red yeast rice powder (batch number: Y202310190) was purchased from Peking University Weixin Biotechnology Co., Ltd.

[0042] Kudzu root extraction: Take 3 kg of kudzu root slices, add 8 times the amount (24 L) of water, soak for 2 hours, then boil twice and decoct for 1.5 hours each time. Combine the filtrates, concentrate to obtain an extract, and place in a vacuum drying oven at -0.07 MPa pressure and 70°C for dehydration. After pulverizing, pass through a 40-mesh sieve.

[0043] Mulberry branch extraction: Take 3 kg of mulberry branches, add 8 times the amount of water (24 L), soak for 2 hours, then decoct twice, each time for 1 - 2 hours. Filter, combine the filtrates and concentrate them into an extract. Put the extract into a vacuum drying oven (-0.05 - -0.09 Mpa pressure, dried at 60°C - 80°C), pulverize and sieve through a 40-mesh sieve to obtain mulberry branch extract powder.

[0044] [Experimental instruments]

[0045] Small animal anesthesia machine (Shenhe Aizhong Medical Technology Co., Ltd.); BL-420I animal ventilator (Chengdu Taimeng Software Co., Ltd.); PowerPac TM HC type high-current electrophoresis instrument, Mini- Tetra type vertical protein electrophoresis tank, Mini-Trans- Module type tank transfer system are all purchased from Shanghai Bio-Rad Laboratories Co., Ltd.; MuttiSkan Mk3 microplate reader (Thermo Fisher Scientific Inc., USA); HT7800 transmission electron microscope (Hitachi, Ltd., Japan); Leica UC7 ultramicrotome (Leica Microsystems GmbH, Germany).

[0046] [Experimental animals]:

[0047] SPF grade SD rats, half male and half female, 120 g - 150 g, sourced from Spf (Beijing) Biotechnology Co., Ltd. (Production license: SCXK (Beijing) 2019 - 0010, Animal certificate number: 110324241100966447). The feeding conditions are constant temperature (25 ± 2)°C and relative humidity (60 ± 5)%.

[0048] [Experimental grouping]

[0049] The control group is fed with maintenance feed normally, and the other groups are fed with high-fat feed after 3 days of adaptive feeding; they are 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 according to different modeling and drug administration methods.

[0050] The daily human clinical dosage: for the red yeast rice + kudzu root group, it is 6 g of red yeast rice and 12 g of kudzu root; for the red yeast rice + mulberry branch group, it is 6 g of red yeast rice and 12 g of mulberry branch; for the red yeast rice + kudzu root + mulberry branch group, it is 6 g of red yeast rice, 12 g of kudzu root and 12 g of mulberry branch. On the premise that the component ratios of each group remain unchanged, the daily drug dosage for rats is calculated as 1.8 g (raw drug) / kg for the red yeast rice + kudzu root group, 1.8 g (raw drug) / kg for the red yeast rice + mulberry branch group, and 3.0 g (raw drug) / kg for the red yeast rice + kudzu root + mulberry branch group; the clinical drug administration dose of nicorandil tablets is 15 mg / person / day, and the drug dosage for rats is calculated as 1.5 mg / kg.

[0051] [Animal Modeling Methods]

[0052] The control group was fed a normal diet and water. After four weeks, blood was collected from the inner canthus of the eyes to measure blood lipid levels. Healthy SD rats were acclimatized for one week and then fed a high-fat diet for four weeks to induce hyperlipidemia. After four weeks, blood was collected from the inner canthus of the eyes to measure blood lipid levels. After confirming abnormal blood lipids, they were divided into 6 groups according to cholesterol (TC) values: high-fat feeding group, model group, positive drug group (nicorandil), red yeast rice + kudzu root group, red yeast rice + mulberry twig group, and red yeast rice + kudzu root + mulberry twig group, so that the mean TC values ​​of rats in each group were similar. Preventive treatment was started for 2 weeks.

[0053] Two weeks after prophylactic treatment, rats in groups other than the control group underwent coronary microcirculation disorder model construction as follows: Rats were anesthetized with 4% isoflurane and then intubated. A small animal ventilator was connected to a gas anesthesia machine to maintain the anesthesia. The left chest of the rats was prepared, and the skin was obliquely cut along the 2nd-4th ribs on the left side of the sternum, with blunt dissection of the muscle layer. The 3rd and 4th intercostal spaces were opened to fully expose the heart and aorta. The aortic arch below the thymus was carefully dissected with forceps, and a cotton thread was passed under the aortic arch. The heart was carefully removed from the thorax, and the thread was gently lifted. The aorta was clamped with vascular clamps, and simultaneously, 4 mg / kg of sodium laurate solution was rapidly injected into the apex of the heart using an insulin syringe. After holding the clamp for 20 seconds, the vascular clamps were released, the heart was returned to its original position, 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 condensation. Postoperatively, 40,000 U of penicillin was injected intramuscularly to prevent wound infection.

[0054] The high-fat feeding group also underwent the same treatment, but used an equal amount of physiological saline instead of sodium lauryl solution. After the model was established, the rats in the high-fat feeding group continued to be fed a high-fat diet for 3 weeks before being harvested.

[0055] The control group continued to be fed and watered normally for 3 weeks before samples were collected.

[0056] The rats in the model group continued to be fed a high-fat diet for 3 weeks before being harvested.

[0057] After the rat models in each treatment group were established, they continued to be fed a high-fat diet and received the drugs for 3 weeks before being harvested.

[0058] [test]

[0059] Echocardiography

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

[0061] Hemodynamic testing

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

[0063] Detection of changes in relevant indicators in rat serum

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

[0065] Observation of myocardial ultrastructure

[0066] Twenty-one days post-surgery, rats were anesthetized, and their hearts were harvested, washed with pre-cooled saline, blotted dry with filter paper, and weighed. The cardiac organ index (heart weight / body weight * 100) was calculated. Subsequently, a 1mm sample was taken. 3 Left ventricular and left ventricular tissues were fixed with 3.5% glutaraldehyde, embedded using conventional electron microscopy techniques, and ultrathin sections (50 nm thick) were prepared. After double staining with uranium ester and lead citrate, the ultrastructural changes of rat myocardial microvessels were observed by transmission electron microscopy.

[0067] Myocardial microvessel density detection

[0068] Twenty-one days post-surgery, cardiac tissue was taken, fixed in 10% neutral morin, embedded in paraffin, sectioned, and incubated overnight at 4°C with CD31 antibody. Secondary antibody was added, and the tissue was incubated at room temperature for 1 hour. DAB staining was performed, followed by hematoxylin counterstaining. The tissue was mounted with neutral resin and observed under a microscope. The vascular endothelial cells showed brownish-yellow staining. ImageJ software was used for analysis, and the number of vascular endothelial cells per unit area was calculated, which is the microvessel density (MVD).

[0069] Western blotting for detection of related protein expression

[0070] Rat heart tissues from each group were removed from a -80°C freezer and placed on ice. RIPA lysis buffer and protease and phosphatase inhibitors were added, and the tissues were mechanically homogenized, centrifuged, and the protein concentration of the supernatant was determined using the BCA protein assay. The protein concentration of each sample was adjusted accordingly. Total myocardial tissue protein (10 μg) was separated by SDS-PAGE and then electrotransferred to a PVDF membrane. The protein-containing PVDF membrane was blocked with skim milk powder for 1.5 hours, and then incubated overnight at 4°C with primary antibody. The antibodies used were: 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 at room temperature for 1 hour with goat anti-rabbit IgG HRP (1:5000) or goat anti-mouse IgG HRP (1:5000). Protein band signals were detected using the Amersham ECL system. The protein bands were quantitatively analyzed using ImageJ software.

[0071] [Statistics and Analysis]

[0072] Data based on mean Mean standard deviation (SD) is expressed as mean ± standard deviation. Statistical comparisons between groups were performed using one-way 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-containing composition on body weight and cardiac organ index in rats with coronary microcirculation disorder induced by sodium laurate injection and high-fat diet.

[0075] Table 1 Results of rat body weight and organ index detection

[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 rats in the model group with coronary microcirculation disorders caused by sodium laurate injection and high-fat diet had a certain degree of weight loss; compared with the model group, there was no significant difference in 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 red yeast rice + kudzu root + mulberry branch group showed a trend of increasing weight.

[0079] Compared with the control group, the organ indices of the heart in rats with coronary microcirculation disorders induced by sodium laurate injection combined with high-fat diet in the model group were significantly decreased, indicating the presence of cardiac lesions. Compared with the model group, the organ indices of the heart in rats in the red yeast rice + kudzu root group and the red yeast rice + kudzu root + mulberry twig group were significantly restored.

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

[0081] Table 2 Results of LVEF and LVFS detection in 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 rat model of coronary microcirculation disorder induced by sodium laurate injection and high-fat diet showed a significant decrease in cardiac function-related indicators, including left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), indicating that the rats in the model group had obvious cardiac systolic dysfunction.

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

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

[0088] Table 3 Results of rat hemodynamic testing

[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 rat model of coronary microcirculation disorder induced by sodium laurate injection and high-fat diet showed a significant decrease in heart rate, 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 impairment of cardiac systolic and diastolic function.

[0092] Compared with the rat model of coronary microcirculatory disturbance induced by sodium laurate injection and high-fat diet, administration of red yeast rice + kudzu root and red yeast rice + kudzu root + mulberry twig significantly improved the LV+dp / dtmax and LV-dp / dtmax of rat hearts, and significantly and effectively improved cardiac function in rats with coronary microcirculatory disturbance. Administration of red yeast rice + mulberry twig did not show a significant improvement in hemodynamics.

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

[0094] Table 4 Results of detection 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 rat model of coronary microcirculation disorder induced by sodium laurate injection and high-fat diet showed significant damage to vascular endothelial function, as evidenced by a significant decrease in serum NO and a significant increase in ET-1, VCAM-1 and ANGII levels, indicating abnormal vascular function.

[0098] Compared with the model group, administration of red yeast rice + kudzu root, red yeast rice + kudzu root + mulberry twig significantly increased NO levels and decreased ET-1 and ANGII levels; administration of red yeast rice + kudzu root, red yeast rice + mulberry twig, red yeast rice + kudzu root + mulberry twig significantly decreased VCAM-1 levels; administration of red yeast rice + kudzu root, red yeast rice + kudzu root + mulberry twig significantly decreased ANGII levels.

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

[0100] Table 5 Results of rat microvessel density detection

[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 model group rats with coronary microcirculation disorder induced by sodium laurate injection and high-fat diet showed a significant decrease in myocardial microvessel density (MVD) and obvious microcirculation disorder.

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

[0106] [Immunohistochemical detection of CD31 expression]

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

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

[0109] Morphology of myocardial tissue in rats with microvascular injury is shown in Figure 2 .

[0110] In the control group, the myocardial cells of the rats were tightly arranged, the cell structure was intact, the large and micro blood vessels were intact and there were no obvious lesions or inflammatory infiltration.

[0111] The model group rats showed obvious cardiomyocyte lysis and necrosis, large-area inflammatory infiltration, significant damage to microvascular structure, perivascular edema, and a large amount of contents within the blood vessels.

[0112] Compared with the model group, the red yeast rice + kudzu root group showed improved myocardial inflammatory infiltration and more intact microvascular structure. The red yeast rice + mulberry twig group showed significantly improved myocardial inflammatory infiltration, with slight damage and edema to the microvascular walls. The red yeast rice + kudzu root + mulberry twig group showed significant improvement in myocardial arrangement and cell integrity, significantly reduced inflammatory infiltration, and relatively intact microvascular walls with slight edema. Its effect on improving myocardial tissue morphology showed a certain synergistic effect with the other two treatment groups.

[0113] [Effects on the ultrastructure of rat myocardium]

[0114] Ultrastructure of rat myocardium Figure 3 .

[0115] In the control group, the myocardial endothelial cells of the rats were tightly connected and 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 rats, the tight junctions of myocardial endothelial cells were significantly damaged, the basement membrane structure was discontinuous, vacuolation was present in the blood vessel wall, the lumen was narrowed and deformed, and the perivascular mitochondrial cristae structure was unclear.

[0117] The red yeast rice + kudzu root group, red yeast rice + mulberry twig group, and red yeast rice + kudzu root + mulberry twig group showed improved loss of tight junctions in rat endothelial cells, improved luminal stenosis, reduced perivascular edema, and a small number of vacuolar areas on the blood vessel wall, indicating improved microvascular endothelial cell condition; the perivascular myocardial mitochondrial structure was relatively clear.

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

[0119] Calculate the drug-in-interaction (CDI) coefficient, where CDI = AB / (A×B). AB represents the ratio of the experimental results of the red yeast rice + kudzu root + mulberry twig group to the model group, and A and B represent the ratios of the experimental results of the red yeast rice + mulberry twig group and the red yeast rice + kudzu root group to the model group, respectively.

[0120] When CDI < 1, it indicates a 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 exhibited a synergistic effect in terms of organ index, LVEF, LVFS, LV+dp / dtmax, LV-dp / dtmax, NO and MVD (CDI<1), suggesting that the compound composed of red yeast rice, kudzu root and mulberry twig can work together to synergistically improve the efficacy on myocardial tissue, cardiac function, vasodilatory function and angiogenesis in model rats.

[0123] in conclusion:

[0124] The core pathology of coronary microcirculation disorders includes abnormal microvascular structure, microvascular obstruction, and abnormal microvascular vasomotor function. Studies have shown that clinical coronary microcirculation disorders can be successfully simulated by injecting rats with sodium laurate and feeding them with a high-fat diet. Based on this, the function of red yeast rice composition was studied.

[0125] The results showed that the red yeast rice composition (red yeast rice + kudzu root + mulberry twig) had a clear effect on improving coronary microcirculation disorders, including: 1) improving abnormal microvascular structure (increasing myocardial microvascular density (MVD), reducing perivascular edema, improving luminal stenosis, and improving endothelial cell condition); 2) improving microvascular obstruction (pathological results showed a reduction in microvascular contents); 3) improving abnormal microvascular vasomotor function (increasing NO and decreasing ANGII); 4) improving lipid metabolism disorders; and 5) improving cardiac function (increasing left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), increasing the maximum rate of increase of left ventricular intraventricular pressure (LV+dp / dtmax) and the maximum rate of decrease of left ventricular intraventricular pressure (LV-dp / dtmax). 6) The three components of red yeast rice, kudzu root, and mulberry twig showed a synergistic effect in terms of organ index, LVEF, LVFS, LV+dp / dtmax, LV-dp / dtmax, NO, and MVD (CDI<1), indicating that the compound composed of red yeast rice, kudzu root, and mulberry twig can work together to improve the efficacy of the treatment on myocardial tissue, cardiac function, vasodilation function, and angiogenesis in the model group rats.

[0126] The exemplary embodiments of the present invention have been described above. 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 within the scope of protection of the present invention.

Claims

1. The use of a red yeast rice composition in the preparation of a drug for treating coronary microcirculation disorders, wherein, by weight, the red yeast rice composition comprises 3-12 parts red yeast rice, 9-15 parts mulberry twigs and 10-15 parts kudzu root.

2. The application according to claim 1, wherein, The red yeast rice composition consists of 6 parts red yeast rice, 12 parts mulberry branches and 12 parts kudzu root by weight.

3. The application according to claim 1 or 2, wherein, The drug also includes pharmaceutically acceptable carriers and / or excipients.

4. The application according to claim 3, wherein, Pharmaceutically acceptable carriers and / or excipients include binders, surfactants, lubricants, fillers, or disintegrants.

5. The application according to claim 4, wherein, The surfactant includes a humectant.

6. The application according to claim 3, wherein, Pharmaceutically acceptable carriers include adsorbent carriers.

7. The application according to claim 1 or 2, wherein, The dosage forms of the drug include oral dosage forms.

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