Use of Dipsacoside VI in the preparation of a medicament for preventing and / or treating tendon adhesion

Abstract

The present invention provides the use of dipsacoside VI in the preparation of a drug for preventing and / or treating tendon adhesion, belonging to the field of biomedicine. The present invention has found through research that dipsacoside VI can promote the polarization of macrophages into the M2 type, inhibit the expression of the fibroblast marker α-SMA, and at the same time inhibit the expression of IGFBP-6. Dipsacoside VI can inhibit tendon adhesion through multiple effects of anti-inflammation, inhibition of fiber activation, and inhibition of IGFBP-6 expression, and can be used to prepare a drug for preventing and / or treating tendon adhesion.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the use of asperosaponin VI in the preparation of a drug for preventing and / or treating tendon adhesion. Background Art

[0002] Tendon adhesion refers to the excessive deposition of extracellular matrix mainly composed of collagen between the damaged tendon and the peritendinous tissue due to the fibrotic healing of the damaged tendon after injury or surgery, resulting in adhesion between the tendon and the surrounding tissues, and further affecting the function of the muscle innervated by the tendon. Its etiology is complex, and the main clinical manifestations are pain at the affected site and joint movement dysfunction, and even disability, which is a common problem after tendon surgery. At present, there is no ideal drug for preventing and treating tendon adhesion, and promoting tendon regeneration and reducing tendon adhesion are great clinical challenges.

[0003] During the pathogenesis of adhesion, growth factors, chemokines and cytokines act together to promote the fibrotic microenvironment. Although the current treatment of fibrotic diseases usually targets the inflammatory response, more and more evidence shows that the mechanisms driving fibrosis are not the same as those regulating inflammation. Therefore, anti-inflammatory alone may not necessarily inhibit fibrosis or treat tendon adhesion.

[0004] Dipsacus asperoides C.Y.Cheng et T.M.Ai. is the root of the plant Dipsacus in the family Dipsacaceae and genus Dipsacus, also known as Chuanduan. The chemical components in Dipsacus asperoides are complex and can be roughly divided into alkaloids, volatile oils, saponins, iridoid glycosides and polysaccharides. Asperosaponin VI (ASAVI) is a triterpenoid saponin compound among them, also known as akebia saponin D or dipsacus saponin C, and has a wide range of pharmacological effects, including neuroprotection, prevention of osteoporosis, myocardial protection, analgesia, liver protection and lipid-lowering. The patent application with the application number 202011182857.1 discloses that asperosaponin VI can treat early tendon injury and promote tendon repair, mainly by promoting the tendon lineage differentiation of tendon stem cells and promoting the regular arrangement of fibers continuously. In early tendon injury, the activity and repair ability of homing cells in the tendon are weak, and fibroblasts and inflammatory cells migrate from the periphery and blood vessels of the tendon to the lesion site, resulting in cell infiltration. Subsequently, the cells migrate and proliferate at the injury site, reorganize the tendon matrix and then initiate tendon repair.

[0005] The treatment of tendon adhesion is different from the pathogenesis and treatment methods of early tendon injuries. Fibroblasts in the surrounding tendon sheath and subcutaneous tissue migrate to the damaged tendon to form granulation tissue to cover the damaged tendon, but this way of healing early tendon injuries will instead cause the formation of adhesion tissue around the tendon. In recent years, scholars have proposed that tendon surface cells have a dual effect of maintaining the normal continuity of tendons and promoting tendon adhesion. During tendon healing, interfering with the proliferation of tendon surface cells will be beneficial to reducing the formation of tendon adhesions.

[0006] Currently, there has been no report on the treatment of tendon adhesion with asperosaponin VI. Summary of the Invention

[0007] The purpose of the present invention is to provide the use of asperosaponin VI in the preparation of a drug for preventing and / or treating tendon adhesion.

[0008] The present invention provides the use of asperosaponin VI in the preparation of a drug for preventing and / or treating tendon adhesion.

[0009] Further, the drug is an anti-inflammatory and tissue fibrosis-inhibiting drug.

[0010] Further, the drug is a drug for inhibiting the expression of macrophage marker CD68.

[0011] Further, the drug is a drug for promoting the polarization of macrophages from M1 type to M2 type.

[0012] Further, the drug is a drug for inhibiting the expression of M1 type macrophage marker CD206.

[0013] Further, the drug is a drug for increasing the expression of M2 type macrophage marker iNOS.

[0014] Further, the drug is a drug for inhibiting the expression of fibroblast marker α-SMA.

[0015] Further, the drug is a drug for inhibiting the expression of IGFBP-6.

[0016] Further, the drug is a drug for preventing and / or treating Achilles tendon adhesion.

[0017] The present invention also provides a drug for preventing and / or treating tendon adhesion, which is a drug prepared from asperosaponin VI as an active ingredient and pharmaceutically acceptable excipients or auxiliary components.

[0018] The present invention has achieved the following beneficial effects:

[0019] The present invention has found through research that asperosaponin VI can promote the polarization of macrophages into the M2 type, inhibit the expression of the fibroblast marker α-SMA, and at the same time inhibit the expression of IGFBP-6. Asperosaponin VI can inhibit tendon adhesion through multiple effects of anti-inflammation, inhibition of fibroblast activation, and inhibition of IGFBP-6 expression, and can be used to prepare drugs for preventing and / or treating tendon adhesion.

[0020] Obviously, based on the above content of the present invention, according to the common general technical knowledge and customary means in the art, without departing from the above basic technical idea of the present invention, various other forms of modifications, substitutions or changes can also be made.

[0021] The following is a further detailed description of the above content of the present invention through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. Brief Description of the Drawings

[0022] Figure 1 It is the HE staining result (100X) of pathological observation of the Control group and the model group in rats.

[0023] Figure 2 It is the expression of the total macrophage marker CD68 detected by immunofluorescence; ASAVI: asperosaponin VI; compared with the Control group, **P < 0.01; compared with the model group, ##P < 0.01.

[0024] Figure 3 It is the expression of the M1 macrophage marker CD206 detected by immunofluorescence; ASAVI: asperosaponin VI; compared with the Control group, **P < 0.01; compared with the model group, ##P < 0.01.

[0025] Figure 4 It is the expression of the M2 macrophage marker iNOS detected by immunofluorescence; ASAVI: asperosaponin VI; compared with the Control group, **P < 0.01; compared with the model group, ##P < 0.01.

[0026] Figure 5 It is the expression of the fibroblast marker α-SMA detected by immunofluorescence; ASAVI: asperosaponin VI; compared with the Control group, **P < 0.01; compared with the model group, ##P < 0.01.

[0027] Figure 6 It is the expression of IGFBP-6 detected by immunohistochemical staining; ASAVI: asperosaponin VI; compared with the Control group, **P < 0.01; compared with the model group, #P < 0.05. Detailed implementation mode

[0028] The raw materials and equipment used in the present invention are all known products, obtained by purchasing commercially available products.

[0029] The CAS number of the asperosaponin VI described in the present invention is 39524-08-8, and the structure is:

[0030]

[0031] Example 1. Research on the treatment of tendon adhesion by asperosaponin VI

[0032] I. Experimental method

[0033] (1) Model establishment

[0034] An acute Achilles tendon injury rat model was established by making a transverse cut through the middle of the longitudinal axis of the Achilles tendon. The injured Achilles tendon stump was wrapped with gelatin sponge dipped in normal saline, and then the skin was sutured. Eight days after the operation, the acute Achilles tendon injury model was identified. HE staining was used to observe the healing of the Achilles tendon and the adhesion fibers to verify whether the model of acute Achilles tendon injury rats was successfully established.

[0035] (2) Grouping

[0036] A randomized controlled animal experiment was conducted. Twenty-eight 2-month-old female SD rats (weighing about 200 g, provided with SPF-grade feed, provided by Chengdu Dashuo Biotechnology Co., Ltd., license number: SCXK-2013-24. The relative humidity in the animal room was 55%-70%, and the room temperature was 20-25 °C) were normally fed for 1 week and then randomly divided into a Control group (n = 10), a model group (n = 10), and an ASAVI treatment group (n = 8) by the digital random method. The Control group was normal SD rats, and the model group and the ASAVI treatment group were modeled using the method described in (1). Eight days after modeling, 2 rats were randomly selected from the Control and model groups for HE staining to observe the healing of the Achilles tendon and the adhesion fibers (such as fiber arrangement, cell density, inflammation, granulation tissue hyperplasia, etc.) to confirm whether the animal modeling was successful. Three days after modeling, the ASAVI treatment group was given ASAVI (CAS number: 39524-08-8 in the Chemical Abstracts of the United States, purchased from Shanghai Yuanye Biotechnology Co., Ltd., with a purity of ≥98% detected by high-performance liquid chromatography, batch number: Z18M10L83256) for treatment. After ASAVI was dissolved in normal saline, it was subcutaneously injected into the left leg at a dose of 40 mg / kg for 8 weeks, once every 3 days. When injecting, the skin around the Achilles tendon was lifted, and the drug was injected into the loose connective tissue around the Achilles tendon without causing tendon tissue damage. Twenty-four hours after the last administration, the rats were fasted but not water-deprived, weighed, and then sacrificed by an overdose anesthesia method (intraperitoneal injection of sodium pentobarbital, 150-200 mg / kg).

[0037] (3) Detection method

[0038] After sacrificing the rats, immunofluorescence staining and immunohistochemical staining were used to analyze the polarization of macrophages, the activation of fibroblasts, and the expression changes of IGFBP-6 in each group.

[0039] II. Experimental results

[0040] (1) Pathological observation of model establishment

[0041] HE staining was used to observe the Achilles tendon healing and adhesion fibrosis conditions, and the results are as Figure 1 shown. The surface of the tendon tissue in the Control group was relatively smooth, the gap between the tendon and the peritendinous tissue was clear, the cell arrangement was regular, and there was no infiltration of inflammatory cells; in the model group, the adhesion range of the tendon was large, granulation tissue hyperplasia occurred around the tendon, the boundary between the tendon and the peritendinous tissue was obvious, and there was a significant infiltration of inflammatory factors. It shows that the rat model of acute Achilles tendon injury established by the present invention was successfully established.

[0042] (2) Changes in macrophage polarization, fibroblast activation and IGFBP-6

[0043] Figure 2 shows the expression of the macrophage marker CD68 in each group, Figure 3 shows the expression of the M1 macrophage marker CD206 in each group, Figure 4 shows the expression of the M2 macrophage marker iNOS in each group. It can be Figures 2 - 4 seen that during the process of adhesion formation, the proportion of M1 macrophages increases. After treatment with asperosaponin VI, the proportion of M1 macrophages can be reduced, promoting the polarization of macrophages to the M2 type, indicating that asperosaponin VI can inhibit the inflammatory response.

[0044] Figure 5 shows the expression of the fibroblast marker α-SMA in each group. It can be Figure 5 seen that during the process of adhesion formation, the activation of the fibroblast marker α-SMA is enhanced, and treatment with asperosaponin VI can inhibit fibroblast activation and thus inhibit adhesion.

[0045] Figure 6 shows the expression of IGFBP-6 in each group. It can be Figure 6 seen that during the process of adhesion formation, the expression of IGFBP-6 increases, and after treatment with asperosaponin VI, the expression of insulin-like growth factor-binding protein 6 (IGFBP-6) is significantly reduced. It shows that asperosaponin VI inhibits adhesion formation by inhibiting the expression of IGFBP-6.

[0046] In summary, the present invention has found through research that asperosaponin VI can promote the polarization of macrophages into the M2 type, inhibit the expression of the fibroblast marker α-SMA, and at the same time inhibit the expression of IGFBP-6. Asperosaponin VI can inhibit tendon adhesion through multiple effects of anti-inflammatory, inhibiting fibroblast activation, and inhibiting the expression of IGFBP-6, and can be used to prepare drugs for preventing and / or treating tendon adhesion.

Claims

1. Use of asperosaponin VI in the preparation of a drug for preventing and / or treating tendon adhesion.

2. The use according to claim 1, wherein: The drug has anti-inflammatory effects and inhibits tissue fibrosis.

3. The use according to claim 2, characterized in that: The drug inhibits the expression of macrophage marker CD68.

4. The use according to claim 2, wherein: The drug promotes the polarization of macrophages from M1 type to M2 type.

5. The use according to claim 4, characterized in that: The drug inhibits the expression of M1 macrophage marker CD206.

6. The use according to claim 4, wherein: The drug increases the expression of M2 macrophage marker iNOS.

7. The use according to claim 2, wherein: The drug inhibits the expression of fibroblast marker α-SMA.

8. The use according to claim 1, characterized in that: The drug inhibits the expression of IGFBP-6.

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