阅读说明：本技术 Xct790在制备治疗钙化性主动脉瓣疾病的药物中的应用 (Application of XCT790 in preparing medicine for treating calcified aortic valve diseases ) 是由 胡王兴 王建安 刘先宝 于 2020-03-16 设计创作，主要内容包括：本发明涉及本发明涉及XCT790在制备治疗钙化性主动脉瓣疾病的药物中的应用,以及抑制瓣膜间质细胞向成骨细胞分化的培养方法。在成骨诱导的体外成钙模型下,显示出了XCT790可以抑制瓣膜间质细胞向成骨细胞转变,并证明其通过上调抗氧化蛋白Hmox1来起到保护作用。本发明的有益效果主要体现在：本发明提供了XCT790在制备治疗钙化性主动脉瓣疾病的药物中的应用,并提供了一种通过XCT790处理后可显著改善瓣膜间质细胞的体外成骨分化的方法,进而为钙化性主动脉瓣疾病提供了新的药物选择。(The invention relates to an application of XCT790 in preparing a medicament for treating calcified aortic valve diseases and a culture method for inhibiting differentiation from valve interstitial cells to osteoblasts. Under an osteogenesis-induced in vitro calcium formation model, XCT790 is shown to inhibit the conversion of valve interstitial cells to osteoblasts and proves that XCT plays a protective role by up-regulating antioxidant protein Hmox 1. The invention has the following beneficial effects: the invention provides application of XCT790 in preparing a medicament for treating calcified aortic valve diseases, and provides a method for remarkably improving in-vitro osteogenic differentiation of valve interstitial cells after XCT790 treatment, thereby providing a new medicament choice for calcified aortic valve diseases.)

Use of XCT790 in the manufacture of a medicament for the treatment of calcified aortic valve disorders.

2. The use according to claim 1, wherein XCT790 is used for the manufacture of a medicament for interfering with osteogenic differentiation of valve stromal cells.

3. The use according to claim 1, wherein XCT790 is used in the manufacture of a medicament for modulating ERR α expression or activity.

4. A culture method for inhibiting differentiation of valvular stromal cells into osteoblasts, the method comprising: inducing and culturing the human valvular interstitial cells with stable passage in a calcification culture medium containing XCT790 for 5-7 days; the XCT 790-containing calcification medium comprises the following components: 10umol/L XCT790, 10mmol/L beta-glycerophosphate, 100nmol/L dexamethasone, 50ug/mL vitamin C, and the solvent is high-sugar DMEM culture medium containing 10% FBS.

5. The method of claim 4, wherein said induction culture is in CO₂Volume concentration 5%, O₂The mixed gas with the volume concentration of 95 percent.

(I) technical field

The invention relates to an application of XCT790 in preparing a medicament for treating calcified aortic valve diseases and a culture method for inhibiting differentiation from valve interstitial cells to osteoblasts.

(II) background of the invention

Calcific Aortic Valve Disease (CAVD) is a progressive disease of the aortic valve, mainly characterized by fibrous hyperplasia and calcification of the valve leaflets leading to stiffening of the valve, and severe cases may cause aortic stenosis, causing obstruction of the left ventricular outflow tract. In developed countries, CAVD is about 2% in people over 65 years of age and about 4% in people over 85 years of age. From the past, CAVD is considered as a passive degenerative change, and is a process of degrading and hardening a valve by deposition with the age; however, basic studies in recent years have shown that CAVD lesions involve endothelial injury, inflammatory infiltration, lipid deposition, as well as extracellular matrix remodeling, conversion of valve stromal cells to osteogenic phenotype, and the like. However, at present, no effective medicine is available for treating CAVD, and valve replacement is a unique treatment mode, but the valve replacement has limitations, so that the research on the specific pathogenesis of CAVD and the search for a proper target point for early intervention are more and more urgent. A large number of experimental studies prove that differentiation from valve interstitial cells to osteogenic cells plays an important role in valve calcification, and intervention in osteogenic differentiation of valve interstitial cells may be an effective method for treating valve calcification.

Estrogen related receptor alpha (ERR α) is the first reported parthenogenetic nuclear receptor that is widely expressed in vivo and plays an important role in a variety of physiological activities. Although endogenous ligands have not been found, it can co-regulate gene transcription by recruiting some regulatory factors to form a transcription complex, playing an important role in cellular energy metabolism and physiological functions, and thus participating in the pathological processes of some common diseases, such as cancer, obesity, diabetes, etc. ERR α also plays an important role in the regulation of osteoblast differentiation and is currently a potential target for the treatment of osteoporosis. Earlier studies found that knocking out ERR α in valvular stromal cells could reduce differentiation of valvular stromal cells into osteoblasts. These results suggest the possibility of ameliorating calcified aortic valve disorders by drug modulation of ERR α expression or activity.

Disclosure of the invention

The invention aims to provide application of XCT790 in preparing a medicament for treating calcified aortic valve diseases and a culture method for inhibiting differentiation from valve interstitial cells to osteoblasts, and aims to provide a potential medicament selection for calcified aortic valve diseases.

The technical scheme adopted by the invention is as follows:

application of XCT790 in preparing medicine for treating calcified aortic valve diseases is provided.

In particular, XCT790 is used for preparing a medicament for interfering in osteogenic differentiation of valve interstitial cells.

Further, XCT790 is used for preparing a medicament for regulating ERR alpha expression or activity.

XCT790 is a strong specific inverse agonist of ERR alpha, has extremely strong specificity to ERR alpha, and when the concentration is lower than 10uM, XCT790 does not show obvious antagonism to related nuclear receptors with similar structures, such as ERR gamma, ER alpha and the like. However, in an in vitro osteogenesis model induced by osteogenesis, XCT790 is shown to inhibit the transition from valve interstitial cells to osteoblasts and proves that XCT plays a protective role by up-regulating antioxidant protein Hmox 1.

The invention also relates to a culture method for inhibiting differentiation of valve mesenchymal cells into osteoblasts, which comprises the following steps: inducing and culturing the human valvular interstitial cells with stable passage in a calcification culture medium containing XCT790 for 5-7 days; the XCT 790-containing calcification medium comprises the following components: 10umol/L XCT790, 10mmol/L beta-glycerophosphate, 100nmol/L dexamethasone, 50ug/mL vitamin C, and 10% FBS solvent in high-sugar DMEM medium. Experiments show that the XCT790 treated human valve mesenchymal cells are obviously reduced in the transformation to osteoblasts under the condition of a calcification induction culture medium consisting of beta-glycerophosphate, dexamethasone and vitamin C in vitro.

Specifically, the induction culture is carried out in CO₂Volume concentration 5%, O₂The mixed gas with the volume concentration of 95 percent.

The invention has the following beneficial effects: the invention provides application of XCT790 in preparing a medicament for treating calcified aortic valve diseases, and provides a method for remarkably improving in-vitro osteogenic differentiation of valve interstitial cells after XCT790 treatment, thereby providing a new potential medicament selection for calcified aortic valve diseases.

(IV) description of the drawings

Figure 1 is a graph of XCT790 treatment improving calcification indicator protein level changes in human valvular stromal cells under in vitro calcification-inducing conditions: NC stands for the conventional culture condition control group, OM + DMSO stands for the calcification induction group, and OM + XCT790 stands for the XCT790 treatment group.

Figure 2 is a graph of XCT790 treatment improving alkaline phosphatase activity of human valve stromal cells under in vitro calcification-inducing conditions: ctrl represents the control group of the conventional culture conditions, OM + DMSO represents the calcification-induced group, and OM + XCT790 represents the XCT 790-treated group.

Figure 3 is a graph of XCT790 treatment improving the change in calcium nodule formation by human valve stromal cells under in vitro calcification-inducing conditions: ctrl represents the control group of the conventional culture conditions, OM + DMSO represents the calcification-induced group, and OM + XCT790 represents the XCT 790-treated group.

Figure 4 is a graph of XCT790 treatment improving calcium content changes in human valve interstitial cells under in vitro calcification-inducing conditions: ctrl represents the control group of the conventional culture conditions, OM + DMSO represents the calcification-induced group, and OM + XCT790 represents the XCT 790-treated group.

(V) detailed description of the preferred embodiments

The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto: