Sclerostin/Receptor Related Protein 4 and Ginkgo Biloba Extract Alleviates β-Glycerophosphate-Induced Vascular Smooth Muscle Cell Calcification By Inhibiting Wnt/β-Catenin Pathway

Abstract

Background: Abnormal mineral metabolism in patients with chronic kidney disease (CKD) may lead to vascular calcification, which is markedly associated with adverse events, including ischemic cardiac diseases and all-cause cardiovascular mortality. Thus, preventing and treating vascular calcification play an important role in improving the prognosis of CKD patients. Objectives: To investigate the potential functions of sclerostin and low-density lipoprotein receptor-related protein 4 (Lrp4) in alleviating the β-glycerophosphate (β-GP)-induced vascular smooth muscle cell (VSMC) calcification, and the protective effect of Ginkgo biloba extract (GBE). Methods: VSMC were extracted from Sprague-Dawley rat aorta and cultured in medium. The VSMCs were divided into 3 groups: (1) Negative control group, (2) β-GP group, in which the VSMCs were treated with β-GP, and (3) GBE and β-GP group, where the VSMCs were treated with both β-GP and GBE. The calcium nodules within the cells were examined by using Alizarin red S staining. The mRNA expression levels of β-catenin and bone gamma-carboxyglutamic-acid-containing proteins (BGP) were detected by real-time PCR. The protein levels of sclerostin and Lrp4 were determined by Western blot. Results: Alizarin red S staining showed that the VSMCs in β-GP group had a distinct orange-red precipitate when compared with VSMCs in the negative control group, while the orange-red precipitate of the GBE and β-GP group was significantly reduced compared to the β-GP group. Real-time PCR showed that the mRNA levels of β-catenin and BGP in VSMCs of β-GP group were significantly higher than those of the negative control group (p < 0.05); while they were significantly reduced in VSMCs of the GBE and β-GP group (p < 0.05). Western blot results showed that the expression of sclerostin in the β-GP group was significantly higher than that in the control group (p < 0.05), whereas Lrp4 was significantly lower than in control group (p < 0.05). Sclerostin in GBE and β-GP group was significantly reduced (p < 0.05), but Lrp4 was significantly elevated when compared with that of the β-GP group (p < 0.05). Conclusion: β-GP induced VSMC calcification by activating the Wnt/β-catenin signaling pathway. Sclerostin and Lrp4 were involved in β-GP-induced VSMC calcification and play an important role. GBE could alleviate VSMC calcification induced by β-GP through inhibiting the Wnt/β-catenin signaling pathway.

Ginsenoside Rb1 ameliorates CKD‐associated vascular calcification by inhibiting the Wnt/β‐catenin pathway

“…High phosphate was reported to activate WNT/β‐catenin signalling, which promoted VC via directly modulating Runx2 gene expression . Moreover, several studies have demonstrated that the inhibition of the WNT/β‐catenin pathway reduces VC . In this study, the activated Wnt/β‐catenin pathway and enhanced nuclear translocation of β‐catenin were observed in β‐GP‐induced VSMC calcification, which is in accordance with the study by Cai et al…”

“…7 Moreover, several studies have demonstrated that the inhibition of the WNT/β-catenin pathway reduces VC. 10,45 In this study, the activated Wnt/β-catenin pathway and enhanced nuclear translocation of β-catenin were observed in β-GP-induced VSMC calcification, which is in accordance with the study by Cai et al 7 PPAR-γ has been reported to regulate glucolipid metabolism and to determine the differentiation of various cell types. 46,47 Recently, studies have reported that PPAR-γ is involved in vascular inflammation and atherosclerosis.…”

Piperlongumine Attenuates High Calcium/Phosphate-Induced Arterial Calcification by Preserving P53/PTEN Signaling

“…Some extracts from natural herbs, including Ginkgo and Radix Puerariae, have protective effects against vascular calcification ( 37 , 38 ). Ginkgo biloba extract (GBE) is produced from Ginkgo leaves and contains several biologically active substances, making it difficult to isolate the effects of the individual biologically active substances.…”

Entelon (vitis vinifera seed extract) reduces degenerative changes in bovine pericardium valve leaflet in a dog intravascular implant model

“…However, many of these approaches have been either ineffective or have produced unwanted side effects [3]. To overcome this drawback, studies on the complementary treatment of calcification have focused on traditional herbal medicines recently [4][5][6].…”