Use of cardamonin in the treatment of aortic valve calcification
A technology for aortic and calcification diseases, applied in cardiovascular system diseases, drug combinations, active ingredients of ketones, etc., can solve the problems of high perioperative mortality, aggravating the economic burden of patients' families, and poor long-term effects. The effect of inhibiting or delaying valve calcification
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Embodiment 1
[0025] Example 1. Construction of a human valve mesenchymal cell calcification model induced by a commercial osteogenic induction medium
[0026] Human valve mesenchymal cells with a culture density close to 80% were starved overnight with 2% FBS+DMEM high-sugar medium, and then starved with a newly configured osteogenic induction medium (Cyagen, product number: HUXMA-90021). The calcification of the valvular interstitial cells was induced, and alizarin red staining was performed after 21 days of induction to investigate the effects of different treatments on the calcification of the valvular interstitial cells.
Embodiment 2
[0027] Embodiment 2, Cardamonin inhibits calcification of human valve interstitial cells
[0028] The human valve interstitial cell calcification model constructed as in Example 1 above was adopted, and the human valve interstitial cells were divided into 3 groups, respectively control group (untreated, normal medium), OM group (osteogenic medium induction) and OM+CDM group (cardamonin was added after osteogenic medium induction). Different concentrations of cardamonin (cardamonin (CAS: 18956-16-6) purchased from Selleck, catalog number: S3942) were used to treat human valve interstitial cells, and IC50 test, CCK8 determination and Alizarin red staining analysis were performed.
[0029] In order to evaluate the cytotoxicity of CDM to hVIC, IC50 test was carried out, and the results showed that CDM had obvious cytotoxicity to hVIC when the concentration exceeded 10 μM ( figure 1 b). Therefore, 10 μM of CDM was used in all subsequent in vitro experiments. In the detection of ...
Embodiment 3
[0030] Example 3, cardamomine inhibits human valve interstitial cell calcification gene protein expression
[0031]The human valve interstitial cell calcification model constructed as in Example 1 above was adopted, and the human valve interstitial cells were divided into 3 groups, respectively control group (untreated, normal medium), OM group (osteogenic medium induction) and OM+CDM group (cardamonin was added after osteogenic medium induction). Human valve mesenchymal cells were treated with 10 μM cardamonin at a final concentration (final concentration in culture medium), and osteogenesis-related genes RUNX2, ALP, OPN, BMP2, SPP1, TNF-α, COL1A2 (type I collagen) were detected at 24h and 48h ).
[0032] Quantitative qRT-PCR results showed that compared with the control group, the expressions of RUNX2, BMP2, SPP1, TNF-α, and COL1A2 in hVIC were significantly up-regulated in the OM group at 24h and 48h (*p figure 2 a and 2b). Similar to the above, the expression of protein ...
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