Use of andrographolide in treatment of calcific aortic valve disease
A technology of andrographolide and calcification, which is applied to cardiovascular system diseases, medical preparations containing active ingredients, organic active ingredients, etc., can solve the problem of high perioperative mortality, increased financial burden on patients' families, and poor long-term effects Good and other questions
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Embodiment 1
[0023] Example 1. Construction of a human valve mesenchymal cell calcification model induced by a commercial osteogenic induction medium
[0024] Take valvular interstitial cells with a culture density close to 80%, starve them overnight with 2% FBS+DMEM high-sugar culture solution, and then complete the starvation with a newly prepared 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
[0025] Example 2, andrographolide inhibits calcification of human valve interstitial cells
[0026] The human valve interstitial cell calcification model constructed as in Example 1 above was used, and the human valve interstitial cells were divided into 3 groups, namely control group (untreated), OM group (osteogenic medium induction) and OM+AGP group (andrographolide was added after osteogenic medium induction). Human valve interstitial cells were treated with different concentrations of andrographolide (andrographolide was purchased from Selleck, catalog number: S2261), and IC50 test, CCK8 determination and alizarin red staining analysis were performed.
[0027] IC50 results as figure 1 Shown in B. cck8 cell viability detection, the concentration of 10μM andrographolide did not show obvious cytotoxicity, and did not affect the cell morphology ( figure 1 C and 1D). After 21 days of treatment of human valve mesenchymal cells with osteogenic medium, alizarin red staining w...
Embodiment 3
[0028] Example 3, andrographolide inhibits human valve interstitial cell calcification gene protein expression
[0029] The human valve interstitial cell calcification model constructed as in Example 1 above was used, and the human valve interstitial cells were divided into 3 groups, namely control group (untreated), OM group (osteogenic medium induction) and OM+AGP group (andrographolide was added after osteogenic medium induction). Human valve mesenchymal cells were treated with 10 μM final concentration of andrographolide (final concentration in culture medium), and osteogenesis-specific genes RUNX2 and ALPL were detected at 24h and 48h.
[0030] Quantitative qRT-PCR results showed that the expression of ALP and RUNX2 (osteogenic differentiation marker gene) was significantly upregulated by osteogenic medium compared with the control group (*p figure 2 a and 2b), showing a statistically significant difference (#p figure 2 c and 2d). Protein expression was verified in anoth...
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