Application of dipsacus asper saponin VI in preparation of medicine for preventing and / or treating tendon adhesion

Abstract

The invention provides application of dipsacus asper saponin VI in preparation of a medicine for preventing and / or treating tendon adhesion, and belongs to the field of biological medicine. The research finds that the dipsacus asper saponin VI can promote the polarization of macrophages to M2 type, inhibit the expression of a fibroblast marker alpha-SMA, and inhibit the expression of IGFBP-6 at the same time. The dipsacus asper saponin VI can inhibit tendon adhesion through multiple effects of resisting inflammation, inhibiting fiber activation and inhibiting IGFBP-6 expression, and can be used for preparing the medicine for preventing and / or treating tendon adhesion.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and specifically relates to use of dipsaponin VI in preparing a medicine for preventing and / or treating tendon adhesion. Background Art

[0002] Tendon adhesion refers to the excessive deposition of collagen-based extracellular matrix between the tendon and the surrounding tissues due to the fibrotic healing of the damaged tendon after tendon injury or surgery, which causes adhesion between the tendon and the surrounding tissues, thereby affecting the function of the tendon to control the muscle. Its etiology is complex, and the main clinical manifestations are pain at the site of onset and joint movement dysfunction, or even disability. It is a common problem after tendon surgery. At present, there is no ideal drug for the prevention and treatment of tendon adhesion. Promoting tendon regeneration and reducing tendon adhesion are great clinical challenges.

[0003] During the pathogenesis of adhesion, growth factors, chemokines, and cytokines work together to promote a fibrotic microenvironment. Although current treatments for fibrotic diseases usually target inflammatory responses, increasing evidence suggests that the mechanisms that drive fibrosis are not equivalent to those that regulate inflammation. Therefore, anti-inflammatory therapy alone does not necessarily inhibit fibrosis or treat tendon adhesions.

[0004] Dipsacus asper is the root of the plant Dipsacus asper (CYChengetT.M.Ai.) of the Dipsacus genus (Dipsacus) of the Dipsacaceae family, also known as Dipsacus asper. The chemical composition of Dipsacus asper is complex and can be roughly divided into alkaloids, volatile oils, saponins, cyclohexene ether terpene glycosides and polysaccharides. Dipsacus asper saponin VI (ASAVI) is one of the triterpenoid saponin compounds, also known as Akebia saponin D or Dipsacus asper saponin C, with a wide range of pharmacological effects, including neuroprotection, prevention of osteoporosis, myocardial protection, analgesia, liver protection and lipid reduction. The patent application with application number 202011182857.1 discloses that Dipsacus asper saponin VI can treat early tendon injuries and promote tendon repair, mainly by promoting the differentiation of tendon stem cells into the tendon system and promoting continuous and regular arrangement of fibers. 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 of the tendon and blood vessels to the lesion site, leading to cell infiltration. Subsequently, the cells migrate and proliferate at the injury site, reorganizing the tendon matrix and thus initiating tendon repair.

[0005] The treatment of tendon adhesion is different from the pathogenesis and treatment of early tendon injury. The fibroblasts in the tendon sheath and subcutaneous tissue around the injury migrate to the damaged tendon to form granulation tissue to cover the damaged tendon. This healing method of early tendon injury will cause the formation of adhesion tissue around the tendon. In recent years, scholars have proposed that tendon surface cells have the dual effects of maintaining normal tendon continuity and promoting tendon adhesion. In the process of tendon healing, interfering with the proliferation of tendon surface cells will help reduce the formation of tendon adhesion.

[0006] At present, there is no report on the use of Dipsacus asper saponin VI to treat tendon adhesion. Summary of the invention

[0007] The object of the present invention is to provide the use of Dipsacus asper saponin VI in preparing a medicament for preventing and / or treating tendon adhesion.

[0008] The present invention provides use of dipsaponin VI in preparing a medicine for preventing and / or treating tendon adhesion.

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

[0010] Furthermore, the drug is a drug that inhibits the expression of macrophage marker CD68.

[0011] Furthermore, the drug is a drug that promotes the polarization of macrophages from M1 type to M2 type.

[0012] Furthermore, the drug is a drug that inhibits the expression of M1 macrophage marker CD206.

[0013] Furthermore, the drug is a drug that increases the expression of iNOS, a marker of M2 macrophages.

[0014] Furthermore, the drug is a drug that inhibits the expression of fibroblast marker α-SMA.

[0015] Furthermore, the drug is a drug that inhibits the expression of IGFBP-6.

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

[0017] The present invention also provides a medicine for preventing and / or treating tendon adhesion, which is prepared by taking Dipsacus as saponin VI as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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

[0019] The present invention has found that Dipsacus root saponin VI can promote macrophage polarization to M2 type, inhibit the expression of fibroblast marker α-SMA, and inhibit the expression of IGFBP-6. Dipsacus root saponin VI can inhibit tendon adhesion through multiple effects such as anti-inflammation, inhibition of fiber activation and inhibition of IGFBP-6 expression, and can be used to prepare drugs for preventing and / or treating tendon adhesion.

[0020] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0021] The above contents of the present invention are further described in detail below through specific implementation methods in the form of examples. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 HE staining results of pathological observation of rat control group and model group (100X).

[0023] Figure 2 Immunofluorescence was used to detect the expression of total macrophage marker CD68; ASAVI: Dipsacus asper saponin VI; compared with the Control group, **P<0.01; compared with the model group, ##P<0.01.

[0024] Figure 3 Immunofluorescence was used to detect the expression of M1 macrophage marker CD206; ASAVI: Dipsacus asper saponin VI; compared with the Control group, **P<0.01; compared with the model group, ##P<0.01.

[0025] Figure 4 Immunofluorescence was used to detect the expression of iNOS, a marker of M2 macrophages; ASAVI: Dipsacus asper saponin VI; compared with the Control group, **P<0.01; compared with the model group, ##P<0.01.

[0026] Figure 5 Immunofluorescence was used to detect the expression of fibroblast marker α-SMA; ASAVI: Dipsaconin VI; compared with the Control group, **P<0.01; compared with the model group, ##P<0.01.

[0027] Figure 6 Immunohistochemical staining was used to detect the expression of IGFBP-6; ASAVI: Dipsaconate VI; compared with the Control group, **P<0.01; compared with the model group, #P<0.05. DETAILED DESCRIPTION

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

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

[0030]

[0031] Example 1: Study on the Treatment of Tendon Adhesion by Dipsacus Aspergillus Saponin VI

[0032] 1. Experimental Methods

[0033] (1) Modeling

[0034] The Achilles tendon acute injury rat model was established by transversely cutting the middle of the Achilles tendon longitudinal axis, and the skin was sutured after wrapping the injured Achilles tendon stump with gelatin sponge soaked in saline. The Achilles tendon acute injury model was identified 8 days after surgery, and the healing and adhesion fiber of the Achilles tendon were observed by HE staining to verify the success of the Achilles tendon acute injury model rat model.

[0035] (2) Grouping

[0036] Randomized controlled animal experiment. 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 of the animal room was 55%-70%, and the room temperature was 20-25°C) were fed normally for 1 week and then divided into the Control group (n=10), the Model group (n=10), and the ASAVI treatment group (n=8) by digital random method. The Control group consisted of normal SD rats, and the Model group and the ASAVI treatment group were modeled by the modeling method described in (1). Eight days after modeling, 2 rats were randomly selected from the Control group and the Model group for HE staining to observe the healing of the Achilles tendon and the adhesion fiber conditions (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 treated with ASAVI (CAS No.: 39524-08-8, purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity ≥98% by high performance liquid chromatography, batch number: Z18M10L83256). ASAVI was dissolved in physiological saline and injected subcutaneously into the left leg, 40 mg / kg, for 8 weeks, once every 3 days. During the injection, 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. After the last administration, the rats were fasted for 24 hours and water was not allowed. After weighing, the rats were killed by overdose anesthesia (sodium pentobarbital injection, 150-200 mg / kg).

[0037] (3) Detection method

[0038] After the rats were killed, immunofluorescence staining and immunohistochemistry staining were used to analyze the changes in macrophage polarization, fibroblast activation and IGFBP-6 expression in each group.

[0039] 2. Experimental Results

[0040] (1) Modeling and pathological observation

[0041] HE staining was used to observe the healing of the Achilles tendon and the adhesion fibers. Figure 1 As 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 cells were arranged regularly and there was no inflammatory cell infiltration; the tendon adhesion range of the model group was larger, the granulation tissue around the tendon proliferated, the boundary between the tendon and the peritendinous tissue was obvious, and there was significant inflammatory factor infiltration. This shows that the rat model of acute Achilles tendon injury of the present invention was successfully established.

[0042] (2) Macrophage polarization, fibroblast activation and changes in IGFBP-6

[0043] Figure 2 is the expression of macrophage marker CD68 in each group. Figure 3 is the expression of CD206, a marker of M1 macrophages in each group. Figure 4 is the expression of iNOS, a marker of M2 macrophages in each group. Figure 2-4 It can be seen that: in the process of adhesion formation, the proportion of M1 macrophages increases. After treatment with Dipsacus asper saponin VI, the proportion of M1 macrophages can be reduced and the polarization of macrophages to M2 can be promoted, indicating that Dipsacus asper saponin VI can inhibit the inflammatory response.

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

[0045] Figure 6 The expression of IGFBP-6 in each group. Figure 6 It can be seen that: during the process of adhesion formation, the expression of IGFBP-6 increased, and the expression of insulin-like growth factor binding protein 6 (IGFBP-6) was significantly reduced after treatment with Dipsacus asper saponin VI, indicating that Dipsacus asper saponin VI inhibits adhesion formation by inhibiting the expression of IGFBP-6.

[0046] In summary, the present invention found that Dipsacus root saponin VI can promote macrophage polarization to M2 type, inhibit the expression of fibroblast marker α-SMA, and inhibit the expression of IGFBP-6. Dipsacus root saponin VI can inhibit tendon adhesion through multiple effects such as anti-inflammatory, inhibition of fiber activation and inhibition of IGFBP-6 expression, and can be used to prepare drugs for preventing and / or treating tendon adhesion.

Claims

1. Use of Dipsacus asper saponin VI in the preparation of a medicament for preventing and / or treating tendon adhesion.

2. The use according to claim 1, characterized in that: The drug is an anti-inflammatory drug and a drug for inhibiting tissue fibrosis.

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

4. The use according to claim 2, characterized in that: The drug is a drug that promotes the polarization of macrophages from M1 type to M2 type.

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

6. The use according to claim 4, characterized in that: The drug is a drug for increasing the expression of iNOS, a marker of M2 macrophages.

7. The use according to claim 2, characterized in that: The drug is a drug for inhibiting the expression of fibroblast marker α-SMA.

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

9. The use according to any one of claims 1 to 8, characterized in that: The medicine is a medicine for preventing and / or treating Achilles tendon adhesion.

10. A drug for preventing and / or treating tendon adhesion, characterized in that: The invention is a medicine prepared by taking chuanxiong saponin VI as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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