Salvianolic acid and notoginsenoside composition and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, and discloses a salvia miltiorrhiza salvianolic acid and notoginsenoside composition, and the mass ratio of notoginsenoside to salvianolic acid in the composition is 1: 5 or 5: 3 or 3: 5. The invention further discloses application of the composition in preparation of drugs for preventing and treating vasculopathy related to metabolic syndromes of metabolic organs. The composition disclosed by the invention can be used for remarkably relieving liver lipid deposition induced by high-fat diet and reducing abnormal accumulation of white and beige fat; wherein the optimal compatibility ratio is 1: 5, and the mechanism of playing the role is related to down-regulation of the expression of liver lipid transport key protein CD36; the invention provides a new intervention strategy and compatibility basis for preventing and treating vasculopathy related to metabolic syndrome, and provides a precise intervention thought of'combining diseases and syndromes' for the research of early prevention of atherosclerosis.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparation technology, and relates to the composition of tanshinone and notoginseng saponins and their application. Background Technology

[0003] Traditional intervention strategies for vascular diseases mainly revolve around modifiable risk factors, with lipid management playing a central role. However, current intervention strategies still have a series of problems that cannot be ignored: (1) Although intensive lipid-lowering therapy has achieved significant results, epidemiological data show that even when LDL-C is controlled at a very low level (<1.8 mmol / L), patients still face a residual cardiovascular risk of up to 40-50%. (2) Studies show that even in patients who use high-intensity statins combined with PCSK9 inhibitors to lower LDL-C to a very low level, the incidence of cardiovascular events is still around 8-9%. (3) Existing lipid-lowering drugs have limited effects on organ-specific metabolic abnormalities, and some statins may even aggravate hepatic steatosis. Non-alcoholic fatty liver disease (NAFLD) and atherosclerosis (AS) often coexist, both of which are characterized by insulin resistance, oxidative stress, and inflammation, suggesting that systemic imbalance in liver metabolism is an important distal factor driving vascular lesions. Therefore, exploring lipid-regulating and anti-inflammatory drugs that can simultaneously intervene in intrahepatic lipid metabolism and vascular inflammation is of great value for the prevention and treatment of ASCVD.

[0004] Traditional Chinese medicine (TCM) and its active ingredients exhibit unique advantages in the prevention and treatment of cardiovascular and metabolic diseases, targeting multiple sites and pathways. Panax notoginseng and Salvia miltiorrhiza are representative TCM herbs for promoting blood circulation and removing blood stasis, with a long history of use and proven efficacy in the prevention and treatment of cardiovascular and cerebrovascular diseases. Panax notoginseng saponins and tanshinone are two representative cardiovascular protective active components, and studies have confirmed that they possess various pharmacological effects, including improving microcirculation, inhibiting platelet aggregation, antioxidation, and anti-inflammation. However, current research largely focuses on single components, and there is a lack of systematic and in-depth exploration into whether the combined use of these two herbs produces a synergistic effect, especially regarding the intervention effect and mechanism targeting the systemic pathological link of the "liver-vascular axis." Summary of the Invention

[0005] In view of this, the purpose of this invention is to clarify the differences in efficacy between Panax notoginseng saponins and Tanshinone when used alone and in combination, to study and screen the optimal ratio that can exert a more efficient advantage, and to preliminarily explore their potential mechanism of action, thereby providing a new path for developing new anti-atherosclerosis strategies based on the synergistic effect of multiple components of traditional Chinese medicine.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A composition of tanshinone and notoginseng saponins, wherein the mass ratio of notoginseng saponins to tanshinone in the composition is 1:5, 5:3, or 3:5.

[0007] To achieve the above objectives, the present invention also provides the following technical solution: The use of the composition in the preparation of a medicament for the prevention and treatment of vascular lesions associated with metabolic syndrome of metabolic organs.

[0008] Furthermore, the metabolic organ is the liver.

[0009] Furthermore, the drug works by improving the liver's lipid processing capacity.

[0010] Furthermore, the drug works by reducing lipid deposition in the liver.

[0011] Furthermore, the drug works by reducing the abnormal accumulation of white fat; when the mass ratio of notoginsenoside to salvianolic acid in the drug is 1:5, the drug works by reducing the volume of white fat cells.

[0012] Furthermore, when the mass ratio of notoginsenosides to salvianolic acid in the drug is 1:5 or 5:3, the drug also works by reducing the abnormal accumulation of beige fat.

[0013] Furthermore, when the mass ratio of notoginsenoside to salvianolic acid in the drug is 1:5, the drug exerts its effect by downregulating the liver lipid transport protein CD36.

[0014] Furthermore, the vascular lesion is atherosclerotic cardiovascular disease.

[0015] The beneficial effects of the present invention are as follows: the composition of the present invention focuses on improving the lipid processing capacity of metabolic organs (liver, fat). The composition of the present invention can significantly reduce liver lipid deposition induced by high-fat diet and reduce the abnormal accumulation of white and beige fat. The mechanism by which the optimal compatibility ratio of 1:5 exerts the above-mentioned effects is related to the downregulation of the expression of CD36, a key protein in liver lipid transport. This invention shows that the core advantage of the combination of Panax notoginseng saponins and Tanshinone lies in the targeted regulation of lipid metabolism homeostasis at the tissue level, rather than simply reducing blood cholesterol. This invention provides a new intervention strategy and compatibility basis for the prevention and treatment of vascular lesions related to metabolic syndrome. This invention also provides a precise intervention approach that combines disease and syndrome for the early prevention of atherosclerosis. Specifically, for the early prevention of atherosclerosis with metabolic disorders, a specific ratio of Panax notoginseng saponin and tanshinone can be used as a combination strategy.

[0016] Other advantages, objectives, and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination, or may be learned from practice of the invention. The objectives and other advantages of the invention can be realized and obtained through the following description. Attached Figure Description

[0017] To make the objectives, technical solutions, and advantages of the present invention clearer, the preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, wherein: Figure 1 This is a graph showing the body weight and liver weight of mice after 12 weeks of normal or high-fat diet in an embodiment of the present invention. Figure 2 This is a graph showing the blood lipid test results of mice after 12 weeks of normal diet or high-fat diet in an embodiment of the present invention; Figure 3 This is a graph showing the serum inflammatory factor detection results of mice after 12 weeks of normal diet or high-fat diet in an embodiment of the present invention; Figure 4 This is a diagram showing the results of the reduction of liver lipid deposition by notoginsenoside / tanshinone in an embodiment of the present invention; Figure 5 This is a diagram showing the results of Panax notoginseng saponins / tanshinone reducing fat accumulation in an embodiment of the present invention; Figure 6 This is a diagram showing the effect of the combination of Panax notoginseng saponins / tanshinone on reducing the size of adipocytes in an embodiment of the present invention; Figure 7 This is a diagram showing the results of transcriptome sequencing analysis of mouse liver tissue in an embodiment of the present invention. Figure 8 This is a diagram showing the results of liver tissue protein verification of CD36 pathway inhibition in an embodiment of the present invention. Detailed Implementation

[0018] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in the following embodiments are only schematic representations of the basic concept of the present invention. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0019] In this embodiment, C57 background ApoE- / - mice, aged 7-8 weeks, were purchased, acclimatized for 1 week, and then fed a high-fat diet for 12 weeks.

[0020] Drug treatment of mice with atherosclerosis: ApoE- / - mice fed a high-fat diet were injected intraperitoneally with 200 μL PBS suspensions containing different proportions of the drug once a day.

[0021] The following solutions were prepared using physiological saline in different proportions of Panax notoginseng saponins and tanshinone tanshinone phenolic acid, as shown in Table 1.

[0022]

[0023] The effects of the above mixture on atherosclerosis were then compared. After 12 weeks of drug administration, all experimental animals were anesthetized, perfused, sampled, and tested. The tissue samples and test parameters are as follows: Basic data: Weight: Weigh yourself once a week.

[0024] Feed intake: The feed intake of each cage of mice was recorded weekly, and the average daily feed intake of mice was recorded 5 days before sampling.

[0025] b. Lipid level detection, inflammation level detection: Blood was drawn from the eye and collected in anticoagulant tubes and EP tubes for later use. Four lipid parameters (total triglycerides, total cholesterol, high-density cholesterol, and low-density cholesterol) and the expression of inflammatory factors IL-1β, IL-6 and TNF-α were detected.

[0026] Frozen sections of liver: Liver stained with Oil Red O, and the number of stained hepatocytes was compared.

[0027] c. Other samples: Liver, white fat, beige fat, and brown fat were collected from each group of mice.

[0028] This embodiment then compares the effects of different ratios of Panax notoginseng saponins / tanshinone on body weight and liver weight.

[0029] C57-background ApoE- / - mice, aged 7-8 weeks, were purchased and, after one week of acclimatization, were randomly divided into nine groups (normal diet group + saline, high-fat diet group + saline, high-fat diet group + different ratios of Panax notoginseng saponins / tanshinone (5:1, 5:3, 1:1, 3:5, 1:5), high-fat diet group + Panax notoginseng saponins (Q), and high-fat diet group + tanshinone (S)). The mice were fed a normal diet or high-fat diet for 12 weeks, and their body weight and liver tissue weight were measured.

[0030] See attached Figure 1The data included: (A) body weight of mice in each treatment group (high-fat diet + saline (SL), normal diet + saline (ND), high-fat diet + Panax notoginseng saponins / tanshinone (5:1, 5:3, 1:1, 3:5, 1:5), high-fat diet + Panax notoginseng saponins (Q), and high-fat diet + tanshinone (S); and (B) liver weight of mice in each treatment group. Data are expressed as mean ± standard deviation.

[0031] Appendix Figure 1 The results showed that different ratios of Panax notoginseng saponins / tanshinone or Panax notoginseng saponins and tanshinone alone had no significant effect on the body weight and liver tissue weight of mice.

[0032] The following study investigated the effects of different ratios of Panax notoginseng saponins / tanshinone on blood lipids.

[0033] C57 background ApoE- / - mice were fed either a normal diet or a high-fat diet, and were administered intraperitoneal drugs every other day (normal diet group + saline, high-fat diet group + saline, high-fat diet group + different ratios of Panax notoginseng saponins / tanshinone (5:1, 5:3, 1:1, 3:5, 1:5), high-fat diet group + Panax notoginseng saponins (Q), high-fat diet group + tanshinone (S)). After 12 weeks, blood samples were collected for the determination of high-density lipoprotein, low-density lipoprotein, total glycosides, and triglycerides.

[0034] See attached Figure 2 The data included: (A) total cholesterol; (B) triglycerides; (C) high-density lipoprotein (HDL); and (D) low-density lipoprotein (LDL). Data are expressed as mean ± standard deviation. Results showed no significant differences between the drug treatment groups and the saline group.

[0035] The following study investigated the effects of different ratios of Panax notoginseng saponins / tanshinone on serum inflammatory factors.

[0036] C57 background ApoE- / - mice were fed either a normal diet or a high-fat diet, and were administered intraperitoneal drugs every other day (normal diet group + saline, high-fat diet group + saline, high-fat diet group + different ratios of Panax notoginseng saponins / tanshinone (5:1, 5:3, 1:1, 3:5, 1:5), high-fat diet group + Panax notoginseng saponins (Q), high-fat diet group + tanshinone (S)). After 12 weeks, blood was collected and centrifuged, and serum IL-1β, IL-6, and TNF-α levels were measured.

[0037] The results are attached. Figure 3 The drug treatments included: (A) IL-1β; (B) IL-6; and (C) TNF-α. The results showed no significant differences between the drug treatment groups and the saline group.

[0038] The following study investigated the effects of different ratios of Panax notoginseng saponins / tanshinone on lipid deposition in the liver. C57-background ApoE- / - mice were fed a normal diet or a high-fat diet, and were given intraperitoneal injections every other day (normal diet group + saline, high-fat diet group + saline, high-fat diet group + different ratios of Panax notoginseng saponins / tanshinone (5:1, 5:3, 1:1, 3:5, 1:5), high-fat diet group + Panax notoginseng saponins (Q), high-fat diet group + tanshinone (S)). After 12 weeks, the mice were perfused, and the livers were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with Oil Red O for observation of lipid droplets.

[0039] The results are attached. Figure 4 The results included: (A) Oil Red O staining of liver tissue after treatment with different ratios of notoginseng saponins / tanshinone; (B) HE staining of liver tissue after treatment with different ratios of notoginseng saponins / tanshinone; and (C) Statistical analysis of the significance of liver lipid deposition after treatment with different ratios of notoginseng saponins / tanshinone. Data are expressed as mean ± standard deviation. Compared with SL, P<0.05, P<0.01, P<0.001.

[0040] Appendix Figure 4 The results showed that after 12 weeks of normal diet feeding, ApoE- / - mice had less lipid deposition in their livers, while the high-fat diet group showed a large amount of lipid deposition in their liver tissues. Compared with the saline group, treatment of ApoE- / - mice with notoginseng saponins (Q) and salvianolic acid (S) alone significantly reduced lipid deposition in the livers. The ratios of notoginseng saponins to total salvianolic acid of 1:5, 5:3, and 3:5 also significantly reduced lipid deposition, while the ratios of 5:1 and 1:1 did not significantly reduce lipid deposition. These results indicate that both notoginseng saponins and salvianolic acid can reduce fatty liver formation, but different ratios yield different effects. This suggests that the synergistic effect of notoginseng saponins and salvianolic acid in reducing fatty liver formation is closely related to their ratio, indicating that exploring the optimal ratio of notoginseng saponins to salvianolic acid is of great significance for effectively reducing fatty liver formation.

[0041] In this embodiment, ApoE- / - mice were treated with a mixture of notoginseng saponins and salvianolic acid in different proportions. After being administered the drug twice a day for 12 consecutive weeks, the body weight, liver weight, white fat, beige fat, and brown fat of the ApoE- / - mice were measured and statistically analyzed.

[0042] The results are attached. Figure 5Among them: (A) Statistical analysis of body weight in each group; (B) Statistical analysis of liver / body weight ratio in each group; (C) Statistical analysis of white fat / body weight ratio in each group; (D) Statistical analysis of brown fat / body weight ratio in each group; (E) Statistical analysis of beige fat / body weight ratio in each group after 12 weeks of administration to ApoE- / - mice.

[0043] Appendix Figure 5 The results showed that after 12 weeks of treatment with different ratios of notoginseng saponins and salvianolic acid, there was no significant difference in body weight or the proportion of liver to body weight in ApoE- / - mice. Compared with the saline group, treatment with notoginseng saponins (Q) or salvianolic acid (S) alone, at ratios of 1:5, 5:3, 3:5, and 1:1, significantly reduced white fat content in ApoE- / - mice. Furthermore, compared with treatment with notoginseng saponins (Q) or salvianolic acid (S) alone, treatment with notoginseng saponins (Q) and salvianolic acid at ratios of 1:5 and 3:5 resulted in a more significant reduction in white fat content, and the effect was better than that at ratios of 5:3 and 1:1.

[0044] Appendix Figure 5 The results also showed that when ApoE- / - mice were treated with notoginsenosides (Q) or total salvianolic acid (S) alone, and with notoginsenosides / total salvianolic acid ratios of 1:5, 3:5, and 5:3, the beige fat content was significantly reduced. The effects of notoginsenosides / total salvianolic acid ratios of 1:5 and 5:3 were better than those of notoginsenosides (Q) or salvianolic acid (S) alone. There were no significant changes in brown fat content in any of the groups.

[0045] The above results further demonstrate that the combined use of Panax notoginseng saponins and salvianolic acid has a more significant effect on reducing lipids, and that the synergistic effect of the two in reducing lipid production is closely related to their ratio. This further shows that exploring the optimal ratio of Panax notoginseng saponins and salvianolic acid is of great significance for effectively reducing lipid production.

[0046] The effects of the combination of Panax notoginseng saponins and Tanshinone on reducing the size of white adipocytes were also studied.

[0047] ApoE- / - mice were treated with a mixture of notoginseng saponins and salvianolic acid in different proportions for 12 consecutive weeks, followed by HE staining of white adipose tissue and statistical analysis of white adipocyte size. Results are attached. Figure 6 Among them: (A) HE staining results of white adipose tissue after treatment with different ratios of Panax notoginseng saponins / tanshinone; (B) Statistical analysis of the significance of white adipocyte size after treatment with Panax notoginseng saponins / tanshinone.

[0048] Appendix Figure 6The results showed that, compared with the saline group, the reduction in white adipocyte volume was most significant after ApoE- / - mice were treated with a 1:5 ratio of Panax notoginseng saponins / tanshinone, while the other treatment groups showed little difference compared with the saline group.

[0049] As is well known, the functions of white adipose tissue (WAT) extend far beyond energy storage. Healthy WAT is highly plastic, adapting to energy changes through adipocyte hypertrophy (enlargement of existing cells) and proliferation (increase in the number of new cells). Importantly, WAT secretes a series of bioactive substances collectively known as "adipokines," including leptin, adiponectin, resistin, and endothelin. Among these, adiponectin has insulin-sensitizing, anti-inflammatory, and anti-atherosclerotic effects; while leptin, in addition to regulating appetite, also participates in immune regulation and vascular function.

[0050] However, in obese individuals, WAT undergoes pathological remodeling: excessive hypertrophy of adipocytes leads to hypoxia and cell death, triggering macrophage infiltration and chronic low-grade inflammation. The secretion profile of adipokines shifts towards pro-inflammatory levels (elevated leptin, resistin, IL-6, and TNF-α), while anti-inflammatory factors (adiponectin) decrease. This "adipo-tissue dysfunction" is a key link connecting obesity with insulin resistance, systemic inflammation, and vascular disease.

[0051] When the volume of white adipocytes decreases, it can reverse adipose tissue dysfunction and improve its secretory function. Specifically, the secretion spectrum of adipokines shifts in a beneficial direction—an increase in the anti-inflammatory factor adiponectin and a decrease in pro-inflammatory factors. This effectively alleviates systemic chronic low-grade inflammation and insulin resistance, thereby cutting off the core pathological link that obesity drives vascular diseases, and thus exerting a positive protective effect against vascular lesions such as atherosclerosis.

[0052] Therefore, it can be seen that the 1:5 ratio of Panax notoginseng saponins / tanshinone in this embodiment can exert a positive protective effect against vascular lesions such as atherosclerosis while significantly reducing the volume of white fat cells.

[0053] This embodiment also investigated the effect of a 1:5 ratio of notoginsenosides / tanshinone on reducing hepatic lipid transport function.

[0054] Previous data in this embodiment have confirmed that a 1:5 ratio of notoginsenosides to salvianolic acid has a good inhibitory effect on hepatic lipid deposition and lipogenesis. To further elucidate the mechanism of action of the 1:5 ratio of notoginsenosides to salvianolic acid in inhibiting lipid deposition, transcriptome sequencing analysis was performed on mouse liver tissue in this embodiment.

[0055] The results are attached. Figure 7The study included: SL-control group, Q-notoginsenoside group, S-tanshinone group, and QS-notoginsenoside / tanshinone 1:5 mixed group. By comparing differentially expressed gene binding pathways, it was found that the fatty acid transporter CD36, which regulates lipid transport in hepatocytes, was enriched in multiple lipid metabolism pathways with significant differences, suggesting that the 1:5 combination of notoginsenoside / tanshinone may inhibit hepatic lipid deposition by reducing CD36 expression.

[0056] This embodiment also confirms that the 1:5 ratio of Panax notoginseng saponins / tanshinone can inhibit the lipid transport pathway in hepatocytes.

[0057] To further confirm that the 1:5 ratio of notoginsenosides / tanshinone may inhibit hepatic lipid deposition by reducing CD36 expression, total protein was extracted from liver tissues after different treatments, and CD36 protein expression was detected. Results are attached. Figure 8 The results showed that the combination of Panax notoginseng saponins / tanshinone in a 1:5 ratio could significantly reduce CD36 protein expression.

[0058] This embodiment reveals the unique value of Panax notoginseng saponins and Salvia miltiorrhiza in regulating local lipid metabolism in tissues. Although they did not significantly alter circulating blood lipid levels, specific ratios (especially 1:5, 5:3, and 3:5) significantly reduced high-fat diet-induced hepatic lipid deposition and decreased the abnormal accumulation of white and beige adipose tissue. The optimal ratio of 1:5 exerts these effects through a mechanism related to downregulating the expression of CD36, a key protein in hepatic lipid transport. By downregulating CD36 expression, excessive fatty acid uptake is inhibited, alleviating hepatic steatosis. This suggests that the core advantage of combining Panax notoginseng and Salvia miltiorrhiza may lie in targeting and regulating lipid metabolic homeostasis at the tissue level, rather than simply lowering blood cholesterol. This provides a new intervention strategy and compatibility basis for the prevention and treatment of vascular lesions related to metabolic syndrome.

[0059] In summary, this embodiment clarifies the differentiated target of action of the ginsenoside / tanshinone combination: the ginsenoside / tanshinone combination focuses on improving the lipid processing capacity of metabolic organs (liver, fat), and its optimal ratio (1:5) exerts its effect by inhibiting CD36. This embodiment also provides a precise intervention approach combining disease and syndrome: for the early prevention of atherosclerosis against a background of metabolic disorders, a specific ratio of ginsenoside / tanshinone combination can be considered.

[0060] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A composition of tanshinone and notoginseng saponins, characterized in that, The mass ratio of notoginsenosides to salvianolic acid in the composition is 1:5, 5:3, or 3:

5.

2. The use of the composition of claim 1 in the preparation of a medicament for preventing and treating vascular lesions associated with metabolic syndrome of metabolic organs.

3. The application according to claim 2, characterized in that, The metabolic organ in question is the liver.

4. The application according to claim 3, characterized in that, The drug works by improving the liver's ability to process lipids.

5. The application according to claim 4, characterized in that, The drug works by reducing lipid deposition in the liver.

6. The application according to claim 5, characterized in that, The drug works by reducing the abnormal accumulation of white fat; when the mass ratio of notoginsenoside to salvianolic acid in the drug is 1:5, the drug works by reducing the volume of white fat cells.

7. The application according to claim 6, characterized in that, When the mass ratio of notoginsenosides to salvianolic acid in the drug is 1:5 or 5:3, the drug also works by reducing the abnormal accumulation of beige fat.

8. The application according to any one of claims 3-7, characterized in that, When the mass ratio of notoginsenoside to salvianolic acid in the drug is 1:5, the drug exerts its effect by downregulating the liver lipid transport protein CD36.

9. The application according to any one of claims 3-7, characterized in that, The vascular lesions are atherosclerotic cardiovascular diseases.

10. The application according to claim 8, characterized in that, The vascular lesions are atherosclerotic cardiovascular diseases.

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