Application of notoginsenoside in preparation of wound repairing medicine

Abstract

The invention discloses application of notoginsenoside in preparation of a wound repairing medicine, and particularly discloses application of notoginsenoside in preparation of the wound repairing medicine. The notoginsenoside comprises ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1 and notoginsenoside R1. The specific action mechanism of notoginsenoside in promoting wound healing is explored, and notoginsenoside is effectively applied to clinical practice, so that the current technical blank can be filled, and a new treatment way can be provided for wound repair, thereby benefiting the majority of patients.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and particularly to the use of notoginsenosides in the preparation of drugs for wound repair. Background Art

[0002] Panax notoginseng, as one of the important members of the treasure house of traditional Chinese medicine in China, is well-known for the active ingredients extracted from its roots - notoginsenosides. Total notoginsenosides are a complex mixture, which contains a variety of monomeric components with significant biological activities, mainly including ginsenoside Rb1, Rg1, Re, and the unique notoginsenoside R1, etc. These components not only have a wide range of pharmacological activities, but also their respective mechanisms of action are different. For example, ginsenoside Rb1 and Rg1 have been proven to have various biological effects such as enhancing immunity, improving memory function, antioxidant and anti-tumor effects; while notoginsenoside R1 has attracted much attention due to its unique effects in hemostasis and promoting wound healing. In addition, ginsenoside Re also has anti-inflammatory, antioxidant and neuroprotective effects, and it also shows potential value in the prevention and treatment of cardiovascular diseases.

[0003] In recent years, with the in-depth study of modern pharmacology, various pharmacological effects of notoginsenosides have been gradually revealed, including but not limited to cardiovascular protection, anti-inflammatory, anti-tumor, immune regulation, etc. These findings further confirm the scientific basis of Panax notoginseng as a traditional Chinese medicine and promote its application in the modern pharmaceutical field. However, although notoginsenosides have shown good curative effects in the treatment of various diseases, in the prior art, there is still a lack of systematic research and application examples on the special use of notoginsenosides and their related components in wound repair. Summary of the Invention

[0004] The purpose of the present invention is to provide the use of notoginsenosides in the preparation of drugs for wound repair, aiming to reveal the potential value of notoginsenosides in wound repair and is expected to provide a new and more effective treatment option for clinical practice.

[0005] The above technical purpose of the present invention is achieved through the following technical solutions: The use of notoginsenosides in the preparation of drugs for wound repair, the application of notoginsenosides in the preparation of drugs for wound repair.

[0006] Further preferably, the notoginsenoside is ginsenoside Re.

[0007] Further preferably, the notoginsenoside is notoginsenoside R1.

[0008] Further preferably, the notoginsenoside is ginsenoside Rb1.

[0009] Further preferably, the notoginsenoside is ginsenoside Rg1.

[0010] Further preferably, the notoginsenoside is any combination of ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1 or notoginsenoside R1.

[0011] In summary, the present invention has the following beneficial effects: Specific components in notoginsenosides, such as ginsenoside Re and notoginsenoside R1, have shown significant effects in promoting fibroblast proliferation and accelerating wound healing in mice. These components can not only rapidly increase the cell proliferation rate in the early stage but also continuously enhance the wound healing speed in the later stage. Further protein expression studies have shown that ginsenoside Re and notoginsenoside R1 may achieve their repair-promoting effects by activating the STAT3 signaling pathway and its downstream target genes. These findings reveal the potential value of notoginsenosides in wound repair, providing a scientific basis for the development of new wound repair drugs. Utilizing these characteristics of notoginsenosides, the present invention is expected to provide a new and more effective treatment option for clinical use, offering innovative methods and ideas for clinical wound repair. Description of the Drawings

[0012] Figure 1 Showing the effects of different notoginsenoside components on the proliferation of mouse and human dermal fibroblasts; Figure 2 Showing the wound repair effects of different notoginsenoside components on mouse wounds; Figure 3 Showing the effects of ginsenoside Re and notoginsenoside R1 on protein expression after treatment of HDFa cells; Figure 4 Showing the effects of overexpressing HIPK2 on STAT3 phosphorylation and the expression of its target gene-encoded proteins. Detailed Embodiments

[0013] In order to enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below in conjunction with embodiments. Those skilled in the art will understand that the following embodiments are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.

[0014] Example 1 An application of notoginsenoside in the preparation of a wound repair drug, wherein the notoginsenoside is ginsenoside Re.

[0015] Example 2 An application of notoginsenoside in the preparation of a wound repair drug, wherein the notoginsenoside is notoginsenoside R1.

[0016] Example 3 The application of notoginsenoside in the preparation of a wound repair drug, wherein the notoginsenoside is ginsenoside Rb1.

[0017] Example 4 The application of notoginsenoside in the preparation of a wound repair drug, wherein the notoginsenoside is ginsenoside Rg1.

[0018] Example 5 The application of notoginsenoside in the preparation of a wound repair drug, wherein the notoginsenoside is any combination of ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1 or notoginsenoside R1.

[0019] Basic research experiments The proliferation of fibroblasts is extremely important for the wound repair process. To study the effects of different notoginsenosides on fibroblast proliferation, four commercial notoginsenoside compounds, including ginsenoside Rb1, Re, Rg1 and notoginsenoside R1, were purchased from a reagent company. HDFa cells (human dermal fibroblasts) and MDF cells (mouse dermal fibroblasts) in good growth state were seeded in 48-well culture plates and treated with DMSO (dimethyl sulfoxide) and 200 µM concentrations of ginsenoside Rb1, Re, Rg1 and notoginsenoside R1 compounds respectively. The cell proliferation rate was detected by the MTT method at different time points (0, 1, 2, 3, 4 and 5 days). The results showed that there was no significant difference between ginsenoside Rb1 and Rg1 and DMSO treatment in the first 3 days of treatment, and the cell proliferation rate could be increased by about 15% on the 4th and 5th days (as shown in Figure 1 A and Figure 1 B). For HDFa and MDF cells treated with ginsenoside Re and notoginsenoside R1, the cell proliferation rate increased by about 20% on the 1st day, which became more obvious over time. By the 5th day, the cell proliferation rate increased by about 50% (as shown in Figure 1 A and Figure 1 B). These results indicate that different notoginsenoside components have different efficiencies in promoting cell proliferation. Among the four notoginsenoside components tested, ginsenoside Re and notoginsenoside R1 showed stronger effects in promoting cell proliferation.

[0020] Mouse experiments The present invention also detected the effects of ginsenosides Rb1, Re, Rg1 and notoginsenoside R1 on wound healing in a mouse wound model. A full-thickness excisional wound of 0.8 cm × 1.5 cm was created on the back of 8-week-old BALB / c mice to establish a skin wound model. The mice were randomly divided into five groups. The first group was injected with DMSO, the second group was injected with 10 mg / kg ginsenoside Rb1, the third group was injected with 10 mg / kg ginsenoside Rg1, the fourth group was injected with 10 mg / kg ginsenoside Re, and the fifth group was injected with 10 mg / kg notoginsenoside R1. The injection sites were all near the wound, and the injection method was subcutaneous injection. There were 8 mice in each group, and the injection was performed once every 2 days. The wound repair of the mice was evaluated after 7 days and 14 days.

[0021] The results showed that there were no significant differences in the wound repair rates of the three groups of mice injected with DMSO, ginsenoside Rb1 and Rg1 on the 7th day and the 14th day. The wounds healed about 20 - 30% on the 7th day and about 45 - 55% by the 14th day (as Figure 2 shown). For the two groups of mice injected with ginsenoside Re and notoginsenoside R1, the wounds healed 85% - 90% on the 7th day and were completely healed by the 14th day (as Figure 2 shown). These results indicate that ginsenoside Re and notoginsenoside R1 can significantly promote the wound healing rate.

[0022] Protein expression study The effects of ginsenoside Re and notoginsenoside R1 on protein expression after treating HDFa cells were referred to Figure 3 (A: Flow chart of experimental design; B: Volcano plot showing differentially expressed proteins after treatment with ginsenoside Re; C: Volcano plot showing differentially expressed proteins after treatment with notoginsenoside R1; D: Heat map showing the 10 up-regulated proteins and 10 down-regulated proteins with the most obvious expression changes after treatment with ginsenoside Re and notoginsenoside R1; E: Immunoblotting to detect the expression levels of proteins). To study how ginsenoside Re and notoginsenoside R1 promote fibroblast proliferation and mouse wound repair, it was planned to examine the protein expression changes genome-wide after treating HDFa cells with these two compounds, and then find some key differentially expressed proteins and study their roles in this process. First, HDFa cells in good growth state were treated with DMSO and 200 μM concentrations of ginsenoside Re and notoginsenoside R1 for 12 hours, and then the proteins were extracted and enzymatically digested. The obtained peptide segments were labeled with iTRAQ reagents and then subjected to proteomic analysis to discover differentially expressed proteins (as Figure 3 shown in A). The results of mass spectrometry analysis showed that the treatment with ginsenoside Re led to the up-regulation of the expression of 382 proteins and the down-regulation of the expression of 263 proteins (as Figure 3Shown in Figure B). After treatment with notoginsenoside R1, 414 proteins were up-regulated and 387 proteins were down-regulated (as Figure 3 shown in Figure C). Among them, the expression of STAT3 was most significantly up-regulated after treatment with ginsenoside Re and notoginsenoside R1 (as Figure 3 shown in Figure D). It was also observed that the expression of proteins encoded by STAT3 target genes (such as Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF) was also significantly up-regulated (as Figure 3 shown in Figure D). In addition, it was also found that the protein level of HIPK2 was significantly up-regulated after treatment with ginsenoside Re and notoginsenoside R1 (as Figure 3 shown in Figure D).

[0023] To verify the accuracy of the mass spectrometry results, the expressions of STAT3, HIPK2, Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF were detected in HDFa cells treated with DMSO, 200 µM ginsenoside Re, and notoginsenoside R1 (3 replicates for each treatment). Meanwhile, the expression of a down-regulated protein CCN1 (cell communication network factor 1) was detected as a control. The immunoblotting results were consistent with the mass spectrometry analysis results, that is, the expressions of STAT3, HIPK2, Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF were significantly up-regulated in HDFa cells treated with ginsenoside Re and notoginsenoside R1, while the expression of CCN1 was down-regulated (as Figure 3 shown in Figure E). These results verified again that STAT3 and the proteins encoded by its downstream target genes were activated in HDFa cells treated with ginsenoside Re and notoginsenoside R1.

[0024] Overexpression of HIPK2 in HDFa cells would activate STAT3 and up-regulate the expression of proteins encoded by STAT3 target genes, referring to Figure 4 (A: Detection of the expression level of HIPK2 mRNA in HIPK2 overexpressing cells; B: Detection of the phosphorylation level of STAT3 and the protein levels of Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF in HIPK2 overexpressing cells. ***P<0.001). The activation of STAT3 for its target genes requires its phosphorylation and translocation into the nucleus, and HIPK2 is a protein kinase. Therefore, next, it was studied whether overexpression of HIPK2 would promote the phosphorylation of STAT3 and up-regulate the proteins encoded by STAT3 target genes. Based on this, the pCDNA3-Flag-HIPK2 plasmid was transfected into HDFa cells. 48 hours after transfection, 1 / 3 of the cells were taken out, and RNA was extracted to detect HIPK2 mRNA to verify the success of its overexpression (as Figure 4As shown in Figure A). Proteins were extracted from the remaining 2 / 3 of the cells and detected by Western blot for protein expression levels. The results showed that with the overexpression of HIPK2, the phosphorylation level of STAT3 increased significantly (as shown in Figure 4 Figure B). At the same time, the overexpression of HIPK2 also increased the protein levels of Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF (as shown in Figure 4 Figure B). These results indicate that the overexpression of HIPK2 may be the cause of the activation of STAT3 and its target genes.

[0025] Notoginsenosides, as the core active ingredients of Panax notoginseng, have made remarkable progress in the research and application in the field of wound repair in recent years, especially showing breakthrough characteristics in new delivery technologies, exploration of molecular mechanisms, and multi-effect synergistic treatment. The following is a summary of the key functions and innovative applications of the research: 1. Bidirectional regulation of hemostasis and anti-thrombosis to optimize the wound microenvironment Notoginsenosides have a unique bidirectional regulation function. They can not only stop bleeding quickly by activating platelets but also decompose fibrin to prevent thrombosis, thus avoiding the local stasis problem that may be caused by traditional hemostatic drugs. This property enables it to reduce bleeding and prevent venous thrombosis during postoperative wound repair, forming an intelligent repair environment of "stopping bleeding without leaving stasis".

[0026] 2. Innovative delivery systems to improve bioavailability Traditional topical drugs are difficult to penetrate deep into the wound due to the skin barrier. The new microneedle technology combined with a wireless power supply near-infrared light (NIR) system solves this problem. For example: the wireless power supply microneedle system: Loading notoginsenoside R1 (NG-R1) into soluble microneedles can penetrate the stratum corneum and target drug delivery to deep tissues. At the same time, near-infrared light promotes local metabolism and anti-inflammation, significantly improving drug penetration and efficacy. Animal experiments show that this system can improve the wound healing speed, reduce the risk of scarring, and the treatment process does not require external power bondage, being flexible and convenient.

[0027] Composite nanovesicle technology: After the combination of total notoginsenosides (PNS) and nanocarriers, the drug stability and targeting can be enhanced. Studies on diabetic foot ulcer models show that this dosage form can upregulate the expression of vascular endothelial growth factor (VEGF), inhibit inflammatory factors (such as IL-6, TNF-α), and promote angiogenesis and collagen deposition, and the curative effect is significantly better than that of ordinary dosage forms.

[0028] 3. Activate key signaling pathways to promote tissue regeneration Notoginsenosides accelerate wound repair by regulating multiple molecular pathways: (1)STAT3 pathway: In a mouse trauma model, it was found that ginsenoside Re and notoginsenoside R1 could activate the STAT3 transcription factor in fibroblasts, thereby upregulating the expression of STAT3 target genes (such as Cyclin D1, c-Myc, BCL2L1, BIRC5, and VEGF). Based on this, it is speculated that ginsenoside Re and notoginsenoside R1 may promote the proliferation of fibroblasts by regulating the STAT3-mediated signaling pathway. These research results can clearly explain how these two saponins promote wound healing, which will not only greatly promote the understanding of the role of notoginsenosides in wound repair but also provide a strong theoretical basis for the development of new wound treatment drugs based on ginsenoside Re and notoginsenoside R1.

[0029] (2)HIF-1α / VEGF / VEGFR2 pathway: In a postoperative anal fistula model, PNS activates hypoxia-inducible factor HIF-1α, promotes the expression of VEGF and its receptor VEGFR2, increases microvessel density (MVD) and collagen synthesis, and shortens the healing cycle.

[0030] (3)Anti-inflammatory and immunomodulatory: Inhibits the release of pro-inflammatory factors (such as IL-6, IL-2), reduces wound inflammatory response, and at the same time creates favorable conditions for tissue regeneration by regulating the Notch1 signaling pathway to balance angiogenesis and inflammation resolution.

[0031] Multi-effect synergy: Anti-infection, antioxidant, and pro-angiogenesis Notoginsenosides exhibit multi-dimensional synergistic effects in wound repair: Anti-infection and antioxidant: Scavenge free radicals, inhibit bacterial colonization, and reduce the risk of wound infection.

[0032] Promote angiogenesis: By increasing the capillary density in the ischemic area, improving local blood circulation, and accelerating the delivery of nutrients, such as in a cerebral infarction model, the angiogenesis efficiency is increased by 30%.

[0033] Collagen remodeling: Upregulates the expression of collagen I and fibronectin (FN), enhances the mechanical strength of wound tissue, and reduces scar formation.

[0034] 4. Precise adaptation for complex wounds Studies have shown that notoginsenoside preparations can be flexibly adapted to various wound types: Diabetic foot ulcer: By regulating the blood glucose and wound microenvironment simultaneously, it improves the healing disorder under hyperglycemic conditions.

[0035] Chronic infectious wound: Such as after anal fistula surgery, it inhibits the excessive inflammation caused by pathogens such as Escherichia coli and promotes granulation tissue growth.

[0036] Burns and deep wounds: The microneedle system can adjust the needle body length to meet the needs of wounds with different depths, and combine with phototherapy to achieve personalized treatment.

[0037] The above embodiments are only explanations of the present invention, and they do not limit the present invention. After reading this specification, those skilled in the art can make modifications to this embodiment without creative contributions according to needs, but as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. Use of notoginsenoside in the preparation of a wound repair drug, characterized in that: Application of notoginsenoside in preparing wound repair drugs.

2. Use of notoginsenoside in the preparation of a wound repair drug according to claim 1, characterized in that: The notoginsenoside is ginsenoside Re.

3. Use of notoginsenoside in the preparation of a wound repair drug according to claim 1, characterized in that: The notoginsenoside is notoginsenoside R1.

4. Use of notoginsenoside in the preparation of a wound repair drug according to claim 1, characterized in that: The notoginsenoside is ginsenoside Rb1.

5. Use of a notoginsenoside in the preparation of a wound repair drug according to claim 1, wherein: The notoginsenoside is ginsenoside Rg1.

6. Use of notoginsenoside in the preparation of a wound repair drug according to claim 1, characterized in that: The notoginsenoside is any combination of ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1 or notoginsenoside R1.

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