Application of hypericin in preparation of product for treating calcified aortic valve disease

Abstract

The invention discloses application of hypericin in preparation of a product for treating calcified aortic valve diseases, and belongs to the technical field of biological medicines. The present invention screens 260 mitochondrial-related compounds approved by the American Food and Drug Administration (FDA) to identify potential therapeutic agents. We employ a high content cell imaging system to assess the effects of these compounds on the expression levels of PGC-1 [alpha] and Runx2, where hypericin shows the most significant increase in the PGC-1 [alpha] / Runx2 ratio. After 52-week-old mice are subjected to western diet and intraperitoneal injection of 2.5 mg / kg hypericin once every other day, compared with a normal saline intraperitoneal injection group, the calcification degree of the valves of the mice supplemented with the hypericin is remarkably reduced after 16 weeks.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of hypericin in preparing a product for treating calcific aortic valve disease. Background Art

[0002] Calcific aortic valve disease (CAVD) is a common heart valve disease. Currently, the treatment of CAVD mainly relies on surgical valve replacement and transcatheter aortic valve replacement (TAVR), but there is a lack of effective drug intervention to prevent or delay the progression of the disease. Targeted development of drugs to prevent or delay valve calcification is of great significance for the prevention and treatment of CAVD. Summary of the invention

[0003] To explore drug targets, we carefully explored the pathophysiological mechanisms of CAVD. This study revealed the important role of PGC-1α deficiency in aging-related calcific aortic valve disease (CAVD) in humans and mice. Further animal and cell experiments have shown that an important pathogenesis of aging-related CAVD is that aging-induced PGC-1α deficiency activates mitochondrial ROS accumulation, causing chemokine release, macrophage M1 polarization and valvular interstitial cell osteoblastic differentiation, exacerbating aging-induced aortic valve calcification ( Figure 1 ).

[0004] Given the critical role of PGC-1α in mitochondrial biogenesis and the impairment of mitochondrial function in aging-related CAVD found in this study, we initiated a screen of 260 mitochondrial-related compounds approved by the US Food and Drug Administration (FDA) to identify potential therapeutic agents. We used a high-content cell imaging system to evaluate the effects of these compounds on the expression levels of PGC-1α and Runx2. Among them, hypericin showed the most significant increase in the PGC-1α / Runx2 ratio. Hypericin, a natural compound extracted from Hypericum perforatum, also known as St. John's wort, was selected for further in vivo and in vitro studies given its established anti-inflammatory and antioxidant properties. We gave 52-week-old mice a Western diet and intraperitoneal injections of 2.5 mg / kg of hypericin every other day. After 16 weeks, the mice supplemented with hypericin had significantly reduced valve calcification compared with the saline intraperitoneal injection group.

[0005] Experimental results show that the technical solution of the present invention, i.e., supplementation with hypericin, can improve the progression of aortic valve calcification in CAVD patients, and is expected to treat CAVD through simple and economical drugs.

[0006] The present invention provides an application of hypericin in preparing a product for treating calcific aortic valve disease. The chemical formula of the hypericin is C 30 H 16O8.

[0007] Preferably, the product is a medicine.

[0008] More preferably, hypericin is the only active ingredient in the medicament.

[0009] More preferably, the dosage form of the drug is powder, tablet, granule, capsule, solution, emulsion or suspension.

[0010] More preferably, the medicine further contains a medically acceptable carrier.

[0011] The present invention also provides an application of a composition in preparing a product for treating calcific aortic valve disease, wherein the composition contains hypericin.

[0012] Preferably, the product is a medicine.

[0013] More preferably, the dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

[0014] More preferably, the medicine further contains a medically acceptable carrier.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] This program is the first to apply hypericin, a common drug with good safety, to the field of valvular calcification. This program has few toxic and side effects, and also has positive effects on anti-oxidation and anti-aging, providing a new program for the drug treatment of CAVD. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The mechanism of aging-induced aortic valve calcification.

[0018] Figure 2 The drug screening results in Example 1, including A. Schematic diagram of mitochondrial targeted drug screening process; B. Heat map of the fluorescence intensity ratio of PGC-1α to Runx2; C. Scatter plot of the relative fluorescence intensity of PGC-1α and Runx2 after each drug treatment; D. Screening results of hypericin; EF. Effect of hypericin on ALP and Runx2 expression in VICs induced by TNF-α; G. Effect of hypericin on calcium deposition in VICs; H. Effect of hypericin on ApoE in the elderly - / - Treatment model diagram of mice; IJ. Hypericin for the treatment of ApoE in elderly - / - Effects of peak transvalvular flow velocity in mice; K. Representative images of HE and Von Kossa staining of aortic valve; L. Effects of hypericin on ApoE in elderly patients - / - Effects of hypericin on aortic valve thickness and calcium deposition in mice; MN. - / - Effects of Runx2 expression on mouse aortic valve. DETAILED DESCRIPTION

[0019] Example 1

[0020] Given the critical role of PGC-1α in mitochondrial biogenesis and our previous findings on impaired mitochondrial function in aging-associated calcific aortic valve disease (CAVD), we initiated a screen of 260 FDA-approved mitochondrial-related compounds for potential therapeutic agents ( Figure 2 A). High-content cell imaging system was used to evaluate the effects of these compounds on the expression levels of PGC-1α and Runx2 ( Figure 2 B, 2C). Hypericin, diminazene aceturate, ivermectin, crizotinib, and rhodamine 6G were the top five drug candidates, which simultaneously upregulated PGC-1α and downregulated Runx2 expression ( Figure 2 D). Among them, Hypericin showed the most significant increase in the PGC-1α / Runx2 ratio, reaching 4.718. Considering the known anti-inflammatory and antioxidant properties of Hypericin, we selected it for further in vitro and in vivo validation studies.

[0021] Primary human aortic valve mesenchymal cells (VICs) were extracted and cultured in DMEM medium containing 10% fetal bovine serum (FBS). When the density was 70%-80%, TNF-α was given for 12 hours and then Hypericin was treated for 48 hours. Western blotting experiments showed that Hypericin counteracted the upregulation of osteogenic differentiation markers ALP and Runx2 in aortic valve mesenchymal cells (VICs) induced by TNF-α ( Figure 2 E-2F). In addition, Alizarin red staining confirmed that Hypericin treatment reduced the calcium nodule formation of VICs after TNF-α stimulation ( Figure 2 G).

[0022] We used 52-week-old apolipoprotein e (Apoe)-deficient mice and fed them a Western diet for 16 weeks to establish a CAVD model. During the model construction process, 15 of the mice were given 2.5 mg / kg hypericin intraperitoneally as an intervention supplement, once every other day for 16 weeks. The endpoint of the modeling was to detect indicators related to valvular calcification: transvalvular flow velocity, calcification-related proteins (RUNX2 and ALP) and silver nitrate staining. The results showed that compared with the control group, the valvular calcification status of the hypericin intervention group was significantly improved under the background of hyperlipidemia, the calcium salt deposition nodules were smaller, the transvalvular flow velocity was lower, and the expression of calcification-related proteins was less.

[0023] In the above in vivo experiments, aged ApoE was injected intraperitoneally with Hypericin. - / - The mice showed reduced peak transvalvular flow velocity, thinner aortic valve thickness, and reduced calcium deposition ( Figure 2 I-2L), and the expression of Runx2 was also decreased ( Figure 2 M-2N). In conclusion, the above results suggest that Hypericin is a promising potential drug for intervention in calcific aortic valve disease (CAVD).

[0024] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of hypericin in the preparation of products for the treatment of calcific aortic valve disease.

2. The use according to claim 1, characterized in that: The product described is a drug.

3. The use according to claim 2, characterized in that: Hypericin is the only active ingredient in the medicine.

4. The use according to claim 3, characterized in that: The dosage form of the drug is powder, tablet, granule, capsule, solution, emulsion or suspension.

5. The use according to claim 4, characterized in that: The medicine also contains a medically acceptable carrier.

6. Use of a composition in the preparation of a product for treating calcific aortic valve disease, characterized in that: The composition contains hypericin.

7. The use according to claim 6, characterized in that: The product described is a drug.

8. The use according to claim 7, characterized in that: The dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

9. The use according to claim 8, characterized in that: The medicine also contains a medically acceptable carrier.

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