Use of panax notoginseng saponins as an effective ingredient in the preparation of a medicine for preventing and / or treating thrombosis caused by vascular endothelial injury

Abstract

The present application relates to the use of panax notoginseng saponins, in particular to the use of panax notoginseng saponins as an effective component in the preparation of a medicine for preventing and / or treating vascular endothelial injury thrombus. The panax notoginseng saponins have the effects of treating thrombus and promoting angiogenesis, wherein the effect of treating thrombus specifically manifests in increasing the number of red blood cells in the heart of a zebrafish model of vascular endothelial injury thrombus, reducing the incidence of cerebral thrombus, and increasing blood flow velocity.

Description

Technical Field

[0001] This invention relates to the use of total saponins of Panax notoginseng, specifically to the use of total saponins of Panax notoginseng as an active ingredient in the preparation of drugs for the prevention and / or treatment of endothelial thrombosis. Background Technology

[0002] Vascular endothelial cells (VECs) are a layer of flattened cells lining the cardiovascular system, acting as a barrier between the blood vessel wall and the blood. Currently, most scholars believe that VECs not only function as a barrier but are also a highly active metabolic reservoir, playing an antithrombotic role and synthesizing various vasoactive substances. These substances influence multiple functions of the blood vessels, including vasoconstriction, coagulation, anticoagulation, and blood flow, thus playing a regulatory role. Damage to or dysfunction of VECs can cause various diseases and is a key initiating factor in many conditions, particularly cardiovascular and cerebrovascular diseases.

[0003] Yan Yanfang et al. Protective effects of total saponins of Panax notoginseng and their main components on hypoxic injury of vascular endothelial cells [J]. Chinese Journal of Experimental Traditional Medical Formulae, 2002(01):34-37. This study investigated the protective effects of total saponins of Panax notoginseng and their main components on vascular endothelial cell injury using the human umbilical vein endothelial cell line ECV304, which mimics the blood-brain barrier. The possible material basis for the protective effect of total saponins of Panax notoginseng against cerebral ischemia is Rb1, Rg1, and Rd. Literature reports that total saponins of Panax notoginseng have an inhibitory effect on myocardial hypertrophy in mice. Panax notoginseng saponin Rb1 can also inhibit the influx of Ca2+ into cardiomyocytes, increase the activity of the sarcoplasmic reticulum calcium pump in cardiomyocytes, and reduce the amount of free calcium in cells, thereby achieving the effect of inhibiting cardiomyocyte hypertrophy.

[0004] The main component of the main root of Panax notoginseng is total saponins, which contain various monomeric saponins, among which ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1 are more abundant. Ginsenoside Rg1 and notoginsenoside R1 can exert pharmacological effects in vivo, such as antiplatelet aggregation, prevention of thrombosis, and protection of brain nerves. However, there are no definitive reports to date on the application of total Panax notoginseng saponins in the preparation of drugs for the prevention and / or treatment of vascular endothelial thrombosis. Summary of the Invention

[0005] The purpose of this invention is to provide the application of total saponins of Panax notoginseng in the preparation of drugs for the prevention and / or treatment of vascular endothelial injury-induced thrombosis. The total saponins of Panax notoginseng involved in this invention have the effects of treating thrombosis and promoting angiogenesis, and can be used to prepare drugs for the prevention and / or treatment of vascular endothelial injury-induced thrombosis.

[0006] The technical solution of the present invention is as follows: the application of a total saponin composition of Panax notoginseng in the preparation of a drug for the prevention / treatment of endothelial thrombosis, characterized in that the total saponins of Panax notoginseng contain 3% to 14% Panax notoginseng saponin R1, 21% to 60% ginseng saponin Rg1, 2% to 9% ginseng saponin Re, and 9% to 48% ginseng saponin Rb1, wherein the total weight of Panax notoginseng saponin R1, ginseng saponin Rg1, ginseng saponin Re and ginseng saponin Rb1 is not less than 57% of the total weight of active ingredients, and the content of ginseng saponin Rd is not more than 5.6% of the total weight of active ingredients.

[0007] Preferably, the endothelial injury thrombosis is characterized by an increase in the number of cardiac red blood cells.

[0008] Preferably, the endothelial thrombosis is characterized by increased blood flow velocity.

[0009] Preferably, the endothelial injury-induced thrombosis is characterized by a reduced incidence of cerebral thrombosis. The present invention has the following advantages: The present invention expands the drug selectivity for treating endothelial injury thrombosis, provides more drug options for the effective treatment of this disease, and further expands the application scope of the Panax notoginseng total saponin composition of the present invention as a drug for treating thrombosis. Attached Figure Description

[0010] Figure 1 Typical image of red blood cell staining intensity in zebrafish heart after sample processing (the blue dashed box indicates the zebrafish heart as the analysis site).

[0011] Figure 2 The staining intensity of zebrafish heart erythrocytes after sample treatment (compared to the normal control group), ### P < 0.001; compared with the model control group, * P < 0.05, ** P < 0.01, *** P < 0.001).

[0012] Figure 3 Blood flow velocity in zebrafish after sample treatment (compared to the normal control group) ### P < 0.001; compared with the model control group, * P < 0.05, *** P < 0.001). Detailed Implementation

[0013] The following examples provide further illustration, but the present invention is not limited to these examples.

[0014] Example 1: Efficacy evaluation of treatment for zebrafish vascular endothelial injury thrombosis model.

[0015] (1) Experimental animals: Albino strain zebrafish with a melanin allele mutation, aged 5 days (5 dpf) after fertilization, were bred and reproduced naturally in pairs. They were provided by Hangzhou Huante Biotechnology Co., Ltd., and the experimental animal use license number was: SYXK (Zhejiang) 2022-0004.

[0016] (2) Experimental materials: The total saponins of Panax notoginseng 1-4 were all provided by Guangxi Wuzhou Pharmaceutical (Group) Co., Ltd., and were all prepared into a 100mg / mL stock solution with sodium chloride injection and stored at -20℃.

[0017] (3) Model establishment, grouping, and drug administration: Zebrafish of the Albino strain with 5 dpf melanin allele mutation were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well. Panax notoginseng total saponins 1-4 were administered intravenously (dosage shown in Table 1). A normal control group and a model control group were also set up, with a volume of 3 mL per well. Except for the normal control group, all other experimental groups were given ponatinib in water to establish a zebrafish vascular endothelial injury thrombosis model, and treated at 28 ℃ for 18 h.

[0018] (4) Detection indicators: zebrafish were stained with o-anisidine. After staining, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The staining intensity of zebrafish heart red blood cells was analyzed and statistically analyzed. The efficacy of the test product in treating vascular endothelial injury thrombosis was evaluated based on the statistical analysis results of this indicator. 10 zebrafish were randomly selected from each experimental group and recorded under a cardiac blood flow analysis system. The blood flow velocity of the zebrafish was analyzed and statistically analyzed. The efficacy of the test product in improving hemodynamics of vascular endothelial injury thrombosis was evaluated based on the statistical analysis results of this indicator.

[0019] (5) Statistical methods: SPSS 26.0 software was used for statistical analysis. The statistical results are expressed as mean ± SE. P < 0.05 indicates that the difference is statistically significant.

[0020] (6) Experimental results: Compared with the normal control group, the staining intensity of cardiac erythrocytes in the model control group was significantly reduced, which was statistically significant (P < 0.001), indicating that the model was successfully established. Compared with the model group, the staining intensity of cardiac erythrocytes was increased in all three concentration groups of test sample (1) (62.5, 125, and 250 ng / tail) (P < 0.05, P < 0.001, P < 0.001); the staining intensity of cardiac erythrocytes was increased in all three concentration groups of test sample (2) (P < 0.001); the staining intensity of cardiac erythrocytes was increased in all three concentration groups of test sample (3) (P < 0.05, P < 0.01, P < 0.01); and the staining intensity of cardiac erythrocytes was increased in all three concentration groups of test sample (4) (P < 0.05, P < 0.05, P < 0.001). See Table 1 for details. Figure 1 and Figure 2 .

[0021] Table 1. Results of the experiment evaluating the efficacy of the samples in treating endothelial thrombosis (n = 10) Compared with the normal control group, ### P < 0.001; compared with the model control group, * P < 0.05,** P <0.01, *** P < 0.001.

[0022] Compared with the normal control group, the blood flow velocity in the model control group was significantly reduced, showing a statistically significant difference (P < 0.001), indicating that the model was successfully established. Compared with the model group, the blood flow velocity in the test sample (1) 125 ng / tail concentration group was significantly increased compared with the model control group (P < 0.05); the blood flow velocity in the test sample (2) 62.5 ng / tail concentration group was significantly increased compared with the model control group (P < 0.05); the blood flow velocity in the test sample (3) 250 ng / tail concentration group was significantly increased compared with the model control group (P < 0.05); the blood flow velocity in the test sample (4) 125 ng / tail concentration group was significantly increased compared with the model control group (P < 0.05). See Table 2 for details. Figure 3 .

[0023] Table 2. Experimental results evaluating the efficacy of the samples in improving hemodynamics in cases of endothelial injury-induced thrombosis (n = 10) Compared with the normal control group,### P < 0.001; compared with the model control group, * P < 0.05, *** P < 0.001.

[0024] in conclusion: Test samples (1), (2), (3), and (4) all have the effect of improving endothelial thrombosis, specifically by increasing the number of red blood cells in the heart of zebrafish in the endothelial thrombosis model and increasing blood flow velocity.

[0025] A comparison of four Panax notoginseng total saponin samples at the same concentration showed their efficacy in improving endothelial thrombosis. 1. Under the condition of 62.5 ng / tail, there was no significant difference in the staining intensity of cardiac erythrocytes. The absolute values ​​of the staining intensity of cardiac erythrocytes in test samples (2) and (4) were higher than those in test samples (1) and (3).

[0026] 2. Under the condition of 125 ng / tail, the absolute value of the staining intensity of cardiac erythrocytes of test sample (3) was higher than that of test samples (1) and (4).

[0027] 3. Under the condition of 250 ng / tail, the absolute values ​​of cardiac erythrocyte staining intensity of test samples (1) and (3) were similar, while the cardiac erythrocyte staining intensity of test sample (4) was significantly higher than that of test sample (1) (P < 0.05), and the absolute value of cardiac erythrocyte staining intensity of test sample (4) was significantly higher than that of test sample (3). The higher the cardiac erythrocyte staining intensity, the better the effect of improving endothelial injury thrombosis. It is suggested that test sample (2) has the best effect in improving endothelial injury thrombosis, followed by test samples (1) and test sample (4).

[0028] A comparison of four Panax notoginseng total saponin samples at the same concentration showed that they had the following effects on improving the hemodynamics of thrombotic injuries caused by vascular endothelial damage: 1. Under the condition of 62.5 ng / tail, there was no significant difference in blood flow velocity among test samples (1), (3) and (4), but the absolute value of blood flow velocity of test sample (1) was higher than that of test samples (3) and (4); the absolute value of blood flow velocity of test sample (2) was significantly higher than that of test samples (1) and (3), but there was no significant difference. The blood flow velocity of test sample (2) was significantly higher than that of test sample (4) (P < 0.05).

[0029] 2. Under the condition of 125 ng / tail, there was no significant difference in blood flow velocity among test samples (1), (3) and (4), but the absolute value of blood flow velocity of test sample (1) was the highest, followed by test sample (4), and the absolute value of blood flow velocity of test sample (3) was the lowest.

[0030] 3. Under the condition of 250 ng / tail, there was no significant difference in blood flow velocity among test samples (1), (3), and (4). The faster the blood flow velocity, the better the effect on improving the hemodynamics of endothelial injury thrombosis. This suggests that test sample (2) is the most effective in improving the hemodynamics of endothelial injury thrombosis, followed by test samples (1) and (4).

[0031] Although the present invention has been described in detail above with general descriptions, specific embodiments, and experiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. The application of a total saponin composition of Panax notoginseng in the preparation of drugs for the prevention / treatment of endothelial thrombosis, characterized in that, The total saponins of Panax notoginseng contain 3%–14% notoginsenoside R1, 21%–60% ginsenoside Rg1, 2%–9% ginsenoside Re, and 9%–48% ginsenoside Rb1. The total weight of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, and ginsenoside Rb1 shall not be less than 57% of the total weight of the active ingredients, and the content of ginsenoside Rd shall not exceed 5.6% of the total weight of the active ingredients.

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