A pharmaceutical composition for preventing and treating diabetic vascular calcification and a preparation method thereof

Abstract

The present application is applicable to the field of medical technology, and provides a pharmaceutical composition for preventing and treating diabetic vascular calcification and a preparation method thereof, wherein the pharmaceutical composition comprises the following raw materials by mass: 15-25 parts of salvia miltiorrhiza, 9-12 parts of coptis chinensis, 9-12 parts of knotweed, 9-15 parts of rhodiola rosea and 9-12 parts of ebony. The present application uses a pharmaceutical composition prepared by compounding salvia miltiorrhiza, coptis chinensis, knotweed, rhodiola rosea and ebony in proportion, which can prevent and treat diabetic vascular calcification; in addition, it can also effectively improve blood sugar metabolism, and has the function of reducing aortic calcium salt deposition and improving aortic pathological morphology.

Description

Technical Field

[0001] The present application belongs to the field of medical technology, and in particular relates to a pharmaceutical composition for preventing and treating diabetic vascular calcification and a preparation method thereof. Background Art

[0002] Diabetes and its complications seriously endanger human health and are currently a major public health issue of global concern. Among them, vascular complications are the primary cause of high disability and mortality in diabetic patients. Vascular calcification (VC) is one of the most common vascular pathological conditions in diabetes, characterized by abnormal calcium phosphate deposition in the vascular wall. It is an important pathological basis of coronary artery disease and a sign of subclinical atherosclerosis, which can easily induce atherosclerotic plaque rupture and the occurrence of adverse cardiovascular events.

[0003] VC is an independent predictor of all-cause mortality and lifetime cardiovascular disease risk in diabetic patients. According to the location of calcium and phosphate deposition, VC can be divided into intimal calcification and medial calcification. Intimal calcification is closely related to atherosclerotic plaques, mainly occurring in the cerebral arteries, coronary arteries, aorta, renal arteries and other sites, and is associated with lipid deposition, inflammatory response and cell necrosis. Medial calcification often coexists with intimal calcification, or can exist independently of intimal calcification. It mainly occurs in small and medium-sized arteries throughout the body and is associated with increased arterial stiffness, impaired high-flow organ perfusion and limb ischemia. In the past, vascular calcification was often regarded as a feature of vascular aging. In recent years, it has been found that vascular calcification is an active, gene-regulated biological process, similar to bone mineralization. Its core link is the osteogenic phenotype transformation of vascular smooth muscle cells, and involves multiple pathological mechanisms such as inflammation, apoptosis, abnormal calcium and phosphorus metabolism, and extracellular vesicle aggregation. VC in diabetic patients often presents with coexistence of intimal nodular calcification and medial calcification. The lesions are more complex, progress faster, and have a worse prognosis than those in non-diabetic patients. Although the pathogenesis of diabetic vascular calcification has been widely explored, its prevention and treatment plans and targeted therapeutic drugs are still unclear. Calcification further increases the risk of adverse cardiovascular events in diabetic patients and the difficulty of implementing vascular reconstruction. How to effectively prevent and treat vascular calcification has become the focus and difficulty in the research field of diabetes and its vascular complications. Summary of the invention

[0004] The purpose of the embodiments of the present application is to provide a pharmaceutical composition for preventing and treating diabetic vascular calcification, aiming to effectively prevent and treat diabetic vascular calcification, improve blood glucose metabolism, and maintain vascular homeostasis and function.

[0005] The embodiment of the present application is implemented as follows: a pharmaceutical composition for preventing and treating diabetic vascular calcification, the pharmaceutical composition comprising the following raw materials in parts by weight:

[0006] 15-25 parts of Salvia miltiorrhiza, 9-12 parts of Coptis chinensis, 9-12 parts of Polygonum cuspidatum, 9-15 parts of Rhodiola rosea and 9-12 parts of Prunus mume.

[0007] Another object of the present invention is to provide a method for preparing a pharmaceutical composition for preventing and treating diabetic vascular calcification, comprising:

[0008] According to the above formula of the pharmaceutical composition for preventing and treating diabetic vascular calcification, weigh salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony, and set aside;

[0009] The salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony are mixed and crushed, and then alcohol extraction is performed, and the obtained extract is concentrated and refrigerated to obtain a concentrated medicinal solution and a mixed medicinal residue;

[0010] The mixed medicinal residue is soaked in water, boiled, left to stand for refrigeration, filtered and concentrated to obtain a concentrated decoction;

[0011] The concentrated decoction is added to the concentrated medicinal solution, and an alcohol solution is added to the solution for standing and refrigerating. The solution is filtered, the ethanol is recovered under reduced pressure, and the solution is concentrated to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification.

[0012] The embodiment of the present application provides a pharmaceutical composition prepared by compounding salvia miltiorrhiza, coptis chinensis, polygonum cuspidatum, rhodiola rosea and ebony in proportion, which can play a role in preventing and treating diabetic vascular calcification; in addition, it can also effectively improve blood sugar metabolism, reduce aortic calcium salt deposition, and improve aortic pathological morphology. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is a graph showing the experimental results of the effect of the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the examples of the present application on the body weight and food intake of rats;

[0014] Figure 2 This is a graph showing the experimental results of the effect of the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the examples of the present application on the fasting blood glucose level of rats;

[0015] Figure 3 This is a graph showing the experimental results of the effect of the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the examples of the present application on the calcification of the aorta in rats;

[0016] Figure 4 This is an Alizarin red staining image of rat aortic arch tissue using the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the examples of the present application;

[0017] Figure 5 This is a graph showing the experimental results of the effect of the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the examples of the present application on the degree of aortic calcification in rats. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the present application more clearly understood, the present application is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0019] Traditional Chinese medicine has the advantages of multi-component, multi-target and multi-pathway holistic effects, and has achieved remarkable therapeutic effects in the prevention and treatment of major diseases such as metabolic diseases and cardiovascular diseases. Diabetes belongs to the category of "Xiao Ke" in traditional Chinese medicine, and diabetic VC belongs to the variant of Xiao Ke, which is caused by the prolonged and uncured Xiao Ke. It has the law of pathological evolution of "vascular stasis as the root of the disease, blood stasis and toxins as the key link, and deficiency of middle qi as the aggravating factor". Based on its pathogenic characteristics and pathological laws, the use of traditional Chinese medicine for syndrome differentiation and treatment can comprehensively and systematically improve the body state and maintain vascular homeostasis. Based on this, the embodiment of the present application has developed a Chinese medicine compound for the prevention and treatment of diabetic vascular calcification. The compound is compounded by five Chinese herbal medicines: Danshen, Coptis chinensis, Polygonum cuspidatum, Rhodiola rosea, and Plum. During the preparation, the five medicinal pieces of Danshen, Coptis chinensis, Polygonum cuspidatum, Rhodiola rosea, and Plum are weighed according to a specific ratio, washed and dried. Then, the Chinese medicine compound can be prepared by distillation extraction, water extraction, alcohol extraction and the like. When preparing the medicine for preventing and treating diabetic vascular calcification, the Chinese medicine compound composition can be added with pharmaceutically acceptable carrier excipients to prepare the medicine in the corresponding dosage form.

[0020] Among them, pharmaceutically acceptable carrier excipients refer to conventional drug carrier excipients in the pharmaceutical field, which do not produce obvious stimulation to organisms and do not eliminate the biological activity and characteristics of the administered compound, such as: diluents, such as water, etc.; fillers, such as starch, sucrose, etc.; binders, such as cellulose derivatives, alginates, gelatin, polyvinyl pyrrolidone; wetting agents, such as glycerol; disintegrants, such as agar, calcium carbonate and sodium bicarbonate; absorption promoters, such as quaternary ammonium compounds; surfactants, such as hexadecanol; adsorption carriers, such as kaolin and bentonite; lubricants, such as talc, calcium stearate and magnesium stearate and polyethylene glycol, etc. In addition, other excipients, such as flavoring agents and sweeteners, etc., may also be added to the above-mentioned pharmaceutical composition.

[0021] Specifically, the pharmaceutical composition for preventing and treating diabetic vascular calcification provided in the embodiments of the present application comprises the following raw materials in parts by weight:

[0022] 15-25 parts of Salvia miltiorrhiza, 9-12 parts of Coptis chinensis, 9-12 parts of Polygonum cuspidatum, 9-15 parts of Rhodiola rosea and 9-12 parts of Prunus mume.

[0023] Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight:

[0024] 20 parts of Salvia miltiorrhiza, 10 parts of Coptis chinensis, 10 parts of Polygonum cuspidatum, 10 parts of Rhodiola rosea and 10 parts of Plum Blossoms.

[0025] Among them, Danshen is bitter and slightly cold. Efficacy: promoting blood circulation and removing blood stasis, relieving pain by menstruation, clearing the heart and eliminating vexation, cooling blood and eliminating carbuncle. Indications: chest pain, heart pain, abdominal pain, accumulation of lumps, pain caused by heat, restlessness and insomnia, irregular menstruation, dysmenorrhea and amenorrhea, and swelling and pain of sores.

[0026] Among them, Coptis chinensis is bitter and cold in nature. Efficacy: clearing away heat and dampness, purging fire and detoxifying. Indications: damp-heat fullness, vomiting and acid regurgitation, diarrhea, jaundice, high fever and coma, hyperactivity of heart fire, restlessness and insomnia, palpitations and restlessness, vomiting and bleeding due to blood heat, red eyes, toothache, thirst, carbuncle, furuncle.

[0027] Among them, Polygonum cuspidatum is slightly bitter and slightly cold. Efficacy: diuresis and jaundice, clearing away heat and detoxification, dispersing blood stasis and relieving pain, relieving cough and reducing phlegm. Indications: damp-heat jaundice, stranguria, leucorrhea, rheumatic pain, carbuncle, burns, amenorrhea, abdominal mass, traumatic injury, and lung heat cough.

[0028] Among them, Rhodiola rosea is sweet, bitter and flat. Efficacy: replenishing qi and activating blood circulation, dredge meridians and relieve asthma. Indications: qi deficiency and blood stasis, chest pain, hemiplegia due to stroke, fatigue and asthma.

[0029] Among them, black plum tastes sour, astringent, and neutral. Efficacy: astringent to the lungs, astringent to the intestines, promoting the production of body fluids, and calming ascaris. It is mainly used to treat chronic cough due to lung deficiency, chronic diarrhea, dysentery, deficiency heat and thirst, ascariasis, vomiting, and abdominal pain.

[0030] The present application also provides a method for preparing a pharmaceutical composition for preventing and treating diabetic vascular calcification, comprising the following steps:

[0031] Step S1: according to the formula of the pharmaceutical composition for preventing and treating diabetic vascular calcification, weigh salvia miltiorrhiza, coptis root, knotweed, rhodiola rosea and ebony for later use;

[0032] Step S2: mixing and grinding the salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony, and then performing alcohol extraction, and concentrating the obtained extract, standing and refrigerating it to obtain a concentrated medicinal solution and mixed medicinal residues;

[0033] Step S3: soaking the mixed medicinal residue in water, boiling it, standing it for refrigeration, filtering it and concentrating it to obtain a concentrated decoction;

[0034] Step S4: adding the concentrated decoction to the concentrated medicinal solution, adding an alcohol solution, standing and refrigerating, filtering, reducing pressure to recover ethanol and concentrating to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification.

[0035] Furthermore, the Chinese medicine extract obtained by the concentration treatment in step S4 can be made into Chinese patent medicines in the form of oral liquid, granules, syrup, wine, paste, powder injection, etc. for preventing and treating diabetic vascular calcification by using conventional processes and adding conventional pharmaceutical excipients.

[0036] Optionally, the step of mixing and crushing the salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony plum, and then performing alcohol extraction, concentrating the obtained extract, standing and refrigerating it to obtain a concentrated medicinal solution and mixed medicinal residues comprises:

[0037] The salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony are mixed and crushed, and then placed in an extraction tank of a hot reflux circulation extraction concentrator, 70% ethanol 10 times the weight of the medicinal materials is added, and the mixture is soaked for 30-50 minutes, and hot reflux extraction is performed until the ethanol-soluble components are completely leached out, and the ethanol in the extraction tank is colorless. When the extract is concentrated to a liquid volume 10 times that of the original medicine, the liquid medicine is released, filtered, and then placed for static refrigeration treatment to obtain a concentrated liquid medicine and mixed medicinal residues.

[0038] Optionally, the step of soaking and decocting the mixed medicinal residues, standing and refrigerating, filtering and concentrating to obtain a concentrated decoction comprises:

[0039] The mixed medicinal residue is soaked in water for 3 hours, and decocted three times with 30 times, 20 times, and 10 times of the original drug in water, respectively, and the decoction time is 2 hours, 1.5 hours, and 1 hour, respectively, to obtain three decoction liquids; the three decoction liquids are combined and allowed to stand and refrigerate, and the supernatant is extracted, filtered, and concentrated to a liquid volume 10 times that of the original drug to obtain a concentrated decoction liquid.

[0040] Optionally, the step of adding the concentrated decoction to the concentrated medicinal solution, adding an alcohol solution, standing and refrigerating, filtering, reducing pressure to recover ethanol, and concentrating to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification comprises:

[0041] The concentrated decoction is added to the concentrated medicinal liquid, and 95% ethanol is gradually added to make the alcohol content in the combined liquid reach 60%-70%, and then the combined liquid is placed in a static refrigerated storage process. After filtering, decompression and ethanol recovery, and concentration, a pharmaceutical composition for preventing and treating diabetic vascular calcification is obtained.

[0042] Optionally, the method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification further comprises:

[0043] The thick paste is subjected to vacuum spray drying to obtain a bulky dry powder;

[0044] The bulky dry powder is added to auxiliary materials and filled into capsules to obtain a capsule-shaped pharmaceutical composition for preventing and treating diabetic vascular calcification.

[0045] The dry powder obtained above can be made into Chinese patent medicine or health drink in the form of capsules, tablets, pills, buccal tablets, etc. for preventing and treating diabetic vascular calcification by using conventional processes and adding conventional pharmaceutical excipients.

[0046] Optionally, the method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification further comprises:

[0047] Starch and xylitol are added to the obtained concentrated solution, and the concentrated solution is processed into tablets to obtain a tablet-shaped pharmaceutical composition for treating diabetic vascular calcification.

[0048] Optionally, the salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony are mixed and crushed into coarse particles of 10-24 mesh.

[0049] Optionally, the conditions for the static refrigeration treatment are: refrigerating at a temperature of 0-3°C for 24-48 hours.

[0050] The following are examples of the pharmaceutical composition for preventing and treating diabetic vascular calcification. The experimental methods used in the following examples are conventional methods unless otherwise specified; the materials, reagents, etc. used can be obtained from commercial channels unless otherwise specified.

[0051] In addition, it should be noted that the numerical values ​​given in the following examples are as accurate as possible, but those skilled in the art understand that due to unavoidable measurement errors and experimental operation problems, each number should be understood as an approximate number rather than an absolutely accurate value.

[0052] Example 1: Preparation of a medicinal oral liquid decoction for preventing and treating diabetic vascular calcification

[0053] Weigh 20 parts of salvia miltiorrhiza, 10 parts of coptis chinensis, 10 parts of polygonum cuspidatum, 10 parts of rhodiola rosea, and 10 parts of ebony. Wash and dry, mix and grind into 10-24 mesh coarse particles, place in the extraction tank of a hot reflux circulation extraction concentrator, add 70% ethanol 10 times the weight of the medicinal material, soak for 40 minutes, perform hot reflux extraction until the ethanol-soluble components are completely leached (the ethanol in the extraction tank is colorless, and the time is about 4.5 hours), concentrate the extract to 10 times the amount of the original drug, release the concentrated liquid, filter, put in a freezer (0-3°C) for 48 hours, filter again, and set aside.

[0054] Soak the residue in pure water for 3 hours, and decoct three times with 30 times, 20 times, and 10 times the amount of water added to the original drug, respectively, for 2 hours, 1.5 hours, and 1 hour, respectively. Combine the decoctions, refrigerate (0-3°C) for 24 hours, take the supernatant, filter, and concentrate to 10 times the amount of the original drug to obtain the concentrated decoction for later use.

[0055] The concentrated decoction is added to the concentrated medicinal liquid, and ethanol is slowly added to extract the concentrated liquid under hot reflux and stirred, and then 95% ethanol is gradually added to make the alcohol content in the combined liquid reach 60%-70%, refrigerated (0-3°C) for 24 hours, filtered, and the ethanol is recovered under reduced pressure. The medicinal liquid is concentrated to contain 2.66 grams of crude drug per 100 ml, bottled, sterilized at high temperature of 115°C for 30 minutes, cooled, and is the finished product after passing the inspection.

[0056] Example 2: Preparation of a drug capsule for preventing and treating diabetic vascular calcification

[0057] Weigh 20 parts of salvia miltiorrhiza, 10 parts of coptis chinensis, 10 parts of polygonum cuspidatum, 10 parts of rhodiola rosea, and 10 parts of ebony. Wash and dry, mix and grind into 10-24 mesh coarse particles, place in the extraction tank of a hot reflux circulation extraction concentrator, add 70% ethanol 10 times the weight of the medicinal material, soak for 40 minutes, perform hot reflux extraction until the ethanol-soluble components are completely leached (the ethanol in the extraction tank is colorless, and the time is about 4.5 hours), concentrate the extract to 10 times the amount of the original drug, release the concentrated liquid, filter, put in a freezer (0-3°C) for 48 hours, filter again, and set aside.

[0058] Soak the residue in pure water for 3 hours, and decoct three times with 30 times, 20 times, and 10 times the amount of water added to the original drug, respectively, for 2 hours, 1.5 hours, and 1 hour, respectively. Combine the decoctions, refrigerate (0-3°C) for 24 hours, take the supernatant, filter, and concentrate to 10 times the amount of the original drug to obtain the concentrated decoction for later use.

[0059] The concentrated decoction is added to the concentrated medicinal solution, and ethanol is slowly added to extract the concentrated solution under hot reflux and stirred, and then 95% ethanol is gradually added to make the alcohol content in the combined solution reach 60%-70%, refrigerated (0-3°C) for 24 hours, filtered, and the ethanol is recovered under reduced pressure. The medicinal solution is concentrated to a thick paste, and vacuum spray-dried to obtain a fluffy dry powder.

[0060] The dry powder is added with appropriate auxiliary materials, containing 4.87 grams of crude drug, put into capsules, packaged, packed, and the finished product is obtained after passing the inspection.

[0061] Example 3: Preparation of a medicinal lozenge for preventing and treating diabetic vascular calcification

[0062] Weigh 20 parts of salvia miltiorrhiza, 10 parts of coptis chinensis, 10 parts of polygonum cuspidatum, 10 parts of rhodiola rosea, and 10 parts of ebony. Wash and dry, mix and grind into 10-24 mesh coarse particles, place in the extraction tank of a hot reflux circulation extraction concentrator, add 70% ethanol 10 times the weight of the medicinal material, soak for 40 minutes, perform hot reflux extraction until the ethanol-soluble components are completely leached (the ethanol in the extraction tank is colorless, and the time is about 4.5 hours), concentrate the extract to 10 times the amount of the original drug, release the concentrated liquid, filter, put in a freezer (0-3°C) for 48 hours, filter again, and set aside.

[0063] Soak the residue in pure water for 3 hours, and decoct three times with 30 times, 20 times, and 10 times the amount of water added to the original drug, respectively, for 2 hours, 1.5 hours, and 1 hour, respectively. Combine the decoctions, refrigerate (0-3°C) for 24 hours, take the supernatant, filter, and concentrate to 10 times the amount of the original drug to obtain the concentrated decoction for later use.

[0064] The concentrated decoction is added to the concentrated medicinal liquid, and ethanol is slowly added to extract the concentrated liquid by hot reflux and stirred, and then 95% ethanol is gradually added to make the alcohol content in the combined liquid reach 60%-70%, and refrigerated (0-3°C) for 24 hours, filtered, and the ethanol is recovered under reduced pressure to concentrate the medicinal liquid; an appropriate amount of starch and xylitol are added to the concentrated liquid, and the concentrated liquid is prepared into a paste, and then prepared into a sheet according to a conventional method, containing 23.75 grams of crude drug, and packaged and boxed. After passing the inspection, it is a finished product.

[0065] The pharmaceutical composition prepared above was subjected to relevant tests, as follows:

[0066] 1. Materials and methods

[0067] 1.1 Experimental animals: 40 6-week-old male SD rats weighing 150-180 g were provided by Beijing Yongxin Kangtai Technology Development Co., Ltd. All animals were housed in an SPF-level animal room with a temperature of 22±2°C, a humidity of 55±5%, and a 12hr / 12hr light-dark cycle, and were provided with open sterilized ordinary feed and water.

[0068] 1.2 Experimental drugs: ① Streptozotocin (STZ): specification: 100 mg, product number S0130, purchased from Sigma-Aldrich Company, USA; ② Vitamin D3 (VD3): specification: 250 mg product number: C1357, purchased from Sigma-Aldrich Company, USA; ③ Traditional Chinese medicine compound: a medicinal lozenge for preventing and treating diabetic vascular calcification prepared from Salvia miltiorrhiza, Coptis chinensis, Polygonum cuspidatum, Rhodiola rosea, and Prunus mume in a mass ratio of (2:1:1:1:1), i.e., the above-mentioned Example 3.

[0069] 1.3 Animal modeling of diabetic vascular calcification: After SD rats were adaptively fed for 1 week, STZ (30 mg / kg / d) was intraperitoneally injected once a day for 3 days starting from the second week. After 3 days, the tail was cut and blood was collected to test the random blood sugar of the rats. The blood sugar value > 16.7 mmol / L was considered to be a successful modeling of diabetic rats. After the diabetic model was stable for 1 week, VD3 (5.5×10 5 U / kg) was injected intraperitoneally once a day for 4 days. Subsequent intervention was performed after stabilization for 14 days after the VD3 injection.

[0070] 1.4 Animal grouping: Stable diabetic VC model rats were randomly divided into 4 groups, 8 rats in each group, namely model group, low-dose Chinese medicine compound group (1.12 g / kg body weight / day), medium-dose Chinese medicine compound group (4.24 g / kg body weight / day), and high-dose Chinese medicine compound group (8.48 g / kg body weight / day). Another 8 SD rats were taken as blank group. Each group of 8 rats was fed with corresponding diet and gavaged with corresponding drugs or equal volume of distilled water for 4 weeks. The requirements of animal ethics in the "Beijing Laboratory Animal Welfare Ethical Review Guidelines" were strictly followed during the animal experiment.

[0071] 1.5 General observation: Weigh each rat every week. Weigh the remaining feed in each cage every week and calculate the total food intake of the rats in a week. The average weekly food intake of each rat = the total food intake of the rats in the cage / 5. Observe and record the mental state and coat color of the rats every week.

[0072] 1.6 Fasting blood glucose test: The fasting blood glucose levels of rats in each group were tested at the same time every week using radioimmunoassay.

[0073] 1.7 Observation of aortic pathological morphology:

[0074] Aortic calcification area detection: Alizarin red staining was used to evaluate the gross aortic calcification area of ​​rats. After the experiment, the rat aorta (up to the brachiocephalic artery, left common carotid artery and left subclavian artery, down to the iliac artery) was stripped, and the aortic adventitia fat was stripped under a stereomicroscope using micro forceps and micro scissors, and the aorta was stained with alizarin red dye, and the calcified area was stained dark red.

[0075] Aortic tissue morphology observation: Alizarin red staining was used to observe the calcification of aortic arch tissue. The rat aorta was removed, segmented, rinsed twice with PBS, fixed with 4% paraformaldehyde (PFA) at room temperature for 48 h, embedded in paraffin and cut into 5 μm sections for Alizarin red staining. Calcified nodules were stained dark red under an optical microscope.

[0076] 1.8 Detection of aortic calcification: ELISA method was used to detect the calcium content and ALP activity of rat aorta.

[0077] 1.9 Statistical analysis: SPSS 26.0 software was used for statistical analysis. Measurement data were expressed as mean ± standard error. Measurement data between multiple groups were compared with normal distribution and homogeneity of variance using one-way ANOVA test; nonparametric test (Kruskal-Wallis method) was used when normal distribution was not met. Paired t test was used for comparison between two groups, and Mann-Whitney method was used for comparison between multiple groups. P < 0.05 indicated that the data were statistically significant.

[0078] 2. Experimental results

[0079] 2.1 Effects of Chinese herbal compound on the general condition of diabetic rats with vascular calcification

[0080] Figure 1 The experimental results of the effect of Chinese herbal compound on the body weight and food intake of rats. A is the weight change graph, and B is the food intake change graph. Food intake was calculated from the beginning of diabetic vascular calcification modeling. Control is the blank group, Model is the model group, DLTM-L is the low-dose group of Chinese herbal compound, DLTM-M is the medium-dose group of Chinese herbal compound, and DLTM-H is the high-dose group of Chinese herbal compound.

[0081] Combination Figure 1 In terms of food intake, compared with the control group, the food intake of diabetic vascular calcification rats in each group decreased with the extension of observation time. After the intervention of the Chinese medicine compound, the condition of the rats improved slightly, and the food intake tended to be stable. In terms of body weight, compared with the control group rats, the body weight gain rate of the rats in the model group and each drug intervention group was slower, but there was no statistically significant difference in body weight between the model group and each drug intervention group.

[0082] 2.2 Effect of Chinese herbal compound on fasting blood glucose level in diabetic rats with vascular calcification

[0083] Figure 2 The experimental results of the effect of Chinese herbal compound on fasting blood glucose level of rats. Control is the blank group, Model is the model group, DLTM-L is the low-dose group of Chinese herbal compound, DLTM-M is the medium-dose group of Chinese herbal compound, and DLTM-H is the high-dose group of Chinese herbal compound. # Compared with the Control group ## P<0.01, *Compared with Model group *P<0.05, **P<0.01.

[0084] Combination Figure 2 Compared with the control group, the blood glucose of rats in the model group and each Chinese medicine compound intervention group was significantly increased (P < 0.01). Before drug intervention, there was no significant difference in fasting blood glucose between the model group and each drug intervention group. After the Chinese medicine compound intervention began, the blood glucose of each intervention group showed a downward trend. After 4 weeks of administration, the blood glucose of rats in the high-dose Chinese medicine compound group was significantly lower than that in the model group, and the difference was statistically significant (P < 0.05). There was no significant difference in blood glucose between the low-dose and medium-dose groups and the model group (P > 0.05).

[0085] 2.3 Effect of Chinese herbal compound on aortic pathological morphology in diabetic rats with vascular calcification

[0086] Figure 3The results of the effect of the Chinese herbal compound on the calcification of the aorta in rats. Control is the blank group, Model is the model group, DLTM-L is the low-dose group of the Chinese herbal compound, DLTM-M is the medium-dose group of the Chinese herbal compound, and DLTM-H is the high-dose group of the Chinese herbal compound.

[0087] Figure 4 The pictures of the Chinese medicine compound on the aortic arch tissue of rats stained with Alizarin red, the magnifications are 4 times and 10 times respectively. Control is the blank group, Model is the model group, DLTM-L is the low-dose group of the Chinese medicine compound, DLTM-M is the medium-dose group of the Chinese medicine compound, and DLTM-H is the high-dose group of the Chinese medicine compound.

[0088] Combination Figure 3-4 Compared with the blank group, the whole aorta of the rats in the model group showed significant red calcified nodules, and there were large areas of dark red calcium salt deposits in the aortic arch. The aortic calcification of the rats in each Chinese medicine compound intervention group was reduced compared with the model group, and the high-dose group was the most significant.

[0089] 2.4 Effect of Chinese herbal compound on aortic calcification in diabetic rats with vascular calcification

[0090] Figure 5 The results of the effect of the Chinese herbal compound on the degree of aortic calcification in rats. A is the aortic ALP activity of each group, and B is the aortic calcium content of each group. Control is the blank group, Model is the model group, DLTM-L is the low-dose group of the Chinese herbal compound, DLTM-M is the medium-dose group of the Chinese herbal compound, and DLTM-H is the high-dose group of the Chinese herbal compound. # Compared with the Control group ### P<0.001, *Compared with Model group **P<0.01, ***P<0.001.

[0091] Combination Figure 5 Compared with the control group, the aortic calcium content and ALP activity of the rats in the model group were significantly increased (P < 0.001). Compared with the model group, different doses of Chinese herbal compound intervention significantly reduced the aortic calcium content and inhibited the aortic ALP activity of the rats (P < 0.01, P < 0.001).

[0092] In summary, vascular calcification is an important pathological basis for diabetic vascular complications, which can significantly increase the risk of adverse cardiovascular events in diabetic patients and the difficulty of implementing vascular reconstruction. Actively exploring its prevention and treatment strategies has important clinical value. The Chinese medicine compound of the embodiment of the present application can significantly reduce fasting blood glucose in rats, inhibit the deposition of aortic calcium content and the activation of calcification index ALP, and reduce aortic calcium salt deposition. It is suggested that the Chinese medicine compound group has the effect of preventing and treating diabetic vascular calcification and maintaining vascular homeostasis.

[0093] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the present application. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the present application shall be subject to the attached claims.

[0094] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A pharmaceutical composition for preventing and treating diabetic vascular calcification, characterized in that: The pharmaceutical composition is made of the following raw materials in parts by weight: 15-25 parts of Salvia miltiorrhiza, 9-12 parts of Coptis chinensis, 9-12 parts of Polygonum cuspidatum, 9-15 parts of Rhodiola rosea and 9-12 parts of Prunus mume.

2. The pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 1, characterized in that: The pharmaceutical composition is made of the following raw materials in parts by weight: 20 parts of Salvia miltiorrhiza, 10 parts of Coptis chinensis, 10 parts of Polygonum cuspidatum, 10 parts of Rhodiola rosea and 10 parts of Plum Blossoms.

3. A method for preparing a pharmaceutical composition for preventing and treating diabetic vascular calcification, characterized in that: include: According to the formula of the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 1 or 2, weigh salvia miltiorrhiza, coptis root, knotweed, rhodiola rosea and ebony for later use; The salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony are mixed and crushed, and then alcohol extraction is performed, and the obtained extract is concentrated and refrigerated to obtain a concentrated medicinal solution and a mixed medicinal residue; The mixed medicinal residues are soaked in water, boiled, left to stand for refrigeration, filtered and concentrated to obtain a concentrated decoction; The concentrated decoction is added to the concentrated medicinal solution, and an alcohol solution is added to the solution for standing and refrigerating. The solution is filtered, the ethanol is recovered under reduced pressure, and the solution is concentrated to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification.

4. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: The steps of mixing and crushing the salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony plum, and then performing alcohol extraction, concentrating the obtained extract, standing and refrigerating it to obtain a concentrated medicinal solution and mixed medicinal residues include: The salvia miltiorrhiza, coptis chinensis, knotweed, rhodiola rosea and ebony are mixed and crushed, and then placed in an extraction tank of a hot reflux circulation extraction concentrator, 70% ethanol 10 times the weight of the medicinal materials is added, and the mixture is soaked for 30-50 minutes. The hot reflux extraction is performed until the ethanol-soluble components are completely leached out. When the extract is concentrated to a liquid volume 10 times that of the original medicine, the liquid medicine is released, filtered, and then placed in a static refrigeration treatment to obtain a concentrated liquid medicine and mixed medicinal residues.

5. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: The step of soaking the mixed medicinal residue in water, decocting, standing and refrigerating, filtering and concentrating to obtain a concentrated decoction comprises: The mixed medicinal residue is soaked in water for 3 hours, and decocted three times with 30 times, 20 times, and 10 times of the original drug in water, respectively, and the decoction time is 2 hours, 1.5 hours, and 1 hour, respectively, to obtain three decoction liquids; the three decoction liquids are combined and allowed to stand and refrigerate, and the supernatant is extracted, filtered, and concentrated to a liquid volume 10 times that of the original drug to obtain a concentrated decoction liquid.

6. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: The steps of adding the concentrated decoction to the concentrated medicinal solution, adding an alcohol solution, standing and refrigerating, filtering, reducing pressure to recover ethanol, and concentrating to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification include: The concentrated decoction is added to the concentrated medicinal solution, and 95% ethanol is gradually added to make the alcohol content in the combined solution reach 60%-70%, and then the solution is placed in a static refrigeration process, filtered, decompressed to recover the ethanol, and concentrated to obtain a pharmaceutical composition for preventing and treating diabetic vascular calcification.

7. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: Also includes: The pharmaceutical composition for preventing and treating diabetic vascular calcification is subjected to vacuum spray drying to obtain a bulky dry powder; The bulky dry powder is added to auxiliary materials and filled into capsules to obtain a capsule-shaped pharmaceutical composition for preventing and treating diabetic vascular calcification.

8. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: Also includes: Starch and xylitol are added to the obtained medicinal composition for preventing and treating diabetic vascular calcification, and the medicinal composition for treating diabetic vascular calcification in the form of a tablet is obtained through tablet-making.

9. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: The salvia miltiorrhiza, coptis root, polygonum cuspidatum, rhodiola rosea and ebony are mixed and crushed into coarse particles of 10-24 mesh.

10. The method for preparing the pharmaceutical composition for preventing and treating diabetic vascular calcification according to claim 3, characterized in that: The conditions for the static refrigeration treatment are: refrigeration at a temperature of 0-3°C for 24-48 hours.

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