CELL PROLIFERATION
·
2020
·Vol. 54
·期 1
·e12955
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IF 5.753
Jun Ma,Lei Zhu,Zhibin Zhou,Tengfei Song,Lei Yang,Xu Yan,Aimin chen,Tian Wen Ye
Objectives Calcium ion signals are important for osteoclast differentiation. Transient receptor potential vanilloid 6 (TRPV6) is a regulator of bone homeostasis. However, it was unclear whether TRPV6 was involved in osteoclast formation. Therefore, the aim of this study was to evaluate the role of TPRV6 in bone metabolism and to clarify its regulatory role in osteoclasts at the cellular level. Materials and methods Bone structure and histological changes in Trpv6 knockout mice were examined using micro-computed tomography and histological analyses. To investigate the effects of Trpv6 on osteoclast function, we silenced or overexpressed Trpv6 in osteoclasts via lentivirus transfection, respectively. Osteoclast differentiation and bone resorption viability were measured by tartrate-resistant acid phosphatase (TRAP) staining and pit formation assays. The expression of osteoclast marker genes, including cathepsin k, DC-STAMP, Atp6v0d2 and TRAP, was measured by qRT-PCR. Cell immunofluorescence and Western blotting were applied to explore the mechanisms by which the IGF-PI3K-AKT pathway was involved in the regulation of osteoclast formation and bone resorption by Trpv6. Results We found that knockout of Trpv6 induced osteoporosis and enhanced bone resorption in mice, but did not affect bone formation. Further studies showed that Trpv6, which was distributed on the cell membrane of osteoclasts, acted as a negative regulator for osteoclast differentiation and function. Mechanistically, Trpv6 suppressed osteoclastogenesis by decreasing the ratios of phosphoprotein/total protein in the IGF–PI3K–AKT signalling pathway. Blocking of the IGF–PI3K–AKT pathway significantly alleviated the inhibitory effect of Trpv6 on osteoclasts formation. Conclusions Our study confirmed the important role of Trpv6 in bone metabolism and clarified its regulatory role in osteoclasts at the cellular level. Taken together, this study may inspire a new strategy for the treatment of osteoporosis.
研究领域:
骨骼生物学
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细胞信号传导
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分子生物学
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药理学
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DOI
PubMed
高影响因子
iScience
·
2021
·
IF 5.458
Jun Ma,Jiajia Lu,Zhibin Zhou,Nan Lu,Jia He,Lei Zhu,Tianwen Ye
Summary The precise effect of estrogen (E2) on osteoclast function is still poorly understood. The aim of this study was to investigate the potential role of transient receptor potential vanilloid 6 (TRPV6) in E2-mediated osteoclast function and to characterize the relevant underlying mechanisms. Here, we found that Trpv6 is drastically decreased in ovariectomy operation animals and the administration of E2 results in an increased expression of Trpv6 in osteoclasts. In contrast, Trpv6 depletion significantly blocked the inhibitory effects of E2 on bone resorption activity, and silencing Trpv6 alleviated E2-induced osteoclast apoptosis. In addition, we found that E2 regulates the transcription of Trpv6 through ERα, by interacting with C/EBPβ and NF-κB. Chip assay analysis indicated that C/EBPβ regulates Trpv6 transcription by binding to Trpv6 promoter fragments −1,866 nt to −1,761 nt and −2,685 nt to −2,580 nt, whereas NF-κB binds to the −953 nt to −851 nt region. We conclude that TRPV6 has a significant effect on E2-mediated osteoclast function.
研究领域:
骨骼生物学
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内分泌学
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分子生物学
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DOI
PubMed
高影响因子
Food & Function
·
2018
·Vol. 9
·期 4
·2251-2260
·
IF 3.289
Guo Liu,Shengwei Sun,Baoyan Guo,Benchun Miao,Zhen Luo,Zumeng Xia,Danyang Ying,Fei Liu,Bin Guo,Jian Tang,Yong Cao,Jianyin Miao
Casein phosphopeptides (CPPs) have been demonstrated to be calcium chelators. Unfortunately, few studies have been reported on the effects of CPPs on the mechanism of the uptake and absorption of Ca2+ and bone metabolism. In this study, a monomeric peptide fraction isolated by RP-HPLC (F6-1) that possessed high calcium transport capacity in Caco-2 cell monolayers was separated and characterized. The effects of F6-1 on the absorption mechanisms of Ca2+ in a Caco-2 monolayer model and bone metabolism in rats were investigated. F6-1 was isolated by preparative and analytical RP-HPLC. Results for calcium transport suggested that the rates of Ca2+ transportation by F6-1 were approximately 2.57, 2.87 and 2.38 times higher than those in the control group at 30, 60 and 120 min, respectively. Results of ultraviolet (UV) spectroscopy indicated that the intensity of UV absorption changed because of the binding of Ca2+ to F6-1. Analysis of transepithelial electrical resistance (TEER) and the expression of TRPV6 in Caco-2 cells showed that F6-1 was likely to influence the transcellular pathway of intestinal absorption of Ca2+ rather than the paracellular pathway. Furthermore, the F6-1 group (1% Ca, 0.03% F6-1) exhibited increases in serum Ca2+ levels, femur length and femur Ca and decreases in serum alkaline phosphatase (ALP) levels and urinary pyridinoline content in a Sprague-Dawley rat model, which implied that F6-1 was beneficial for bone calcification. Overall, our results suggested that F6-1 enhanced the transport of Ca2+ in Caco-2 cells by affecting the transcellular pathway by upregulating the expression of TRPV6. F6-1 also improved bone formation and prevented bone resorption to benefit bone health in rats, which provided a basis for using F6-1 in calcium supplements or functional foods.
研究领域:
营养学
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生物化学
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食品科学
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生理学
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DOI
PubMed
高影响因子
Foods
·
2026
·Vol. 15
·期 14
·2490
·
IF 6
Yue Tian,Wenting Yang,Yangzheng He,Xin Bi,Yong Sun
Rice protein peptides, abundant byproducts of rice processing, represent a sustainable source for developing novel nutritional delivery systems. To address the low bioavailability of traditional calcium supplements, this study aimed to fabricate a high-performance calcium-chelating complex (RPP-Ca) and elucidate its functional mechanism. The synthesis process was systematically optimized, yielding a maximum calcium-binding capacity of 93.98 ± 1.99 mg/g under optimal conditions (pH 10, 70 °C, 50 min reaction time, peptide-to-calcium mass ratio of 2:1). Physicochemical characterization utilizing scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) confirmed successful chelation, revealing significant microstructural reorganization and enhanced thermal stability compared to native peptides. Functional validation via in vitro Caco-2 cell models and in vivo calcium-deficient mouse models demonstrated that RPP-Ca significantly promotes intestinal calcium absorption and osteogenesis. Mechanistically, these effects were mediated through the activation of the TRPV6-Calbindin9k signaling axis. These findings underscore the potential of industrial rice protein peptides as an effective and bioavailable calcium fortification ingredient, providing a theoretical basis for the high-value utilization of rice byproducts in functional foods.
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2023
·
IF 8.8
Zheng Fang,Xiaoping Wu,Fangfang Wang,Fan Li,Xixi Cai,Vincent Guyonnet,Shaoyun Wang
A self-assembled peptides-calcium-Vitamin D 3 ternary delivery system (CSPH-Ca-VD 3 ) was prepared to investigate the promotion of cellular calcium transport. The constructed CSPH-Ca-VD 3 nanocomplex exhibited a spherical structure with a size of 135.2 ± 10.2 nm. Based on the thermodynamic calculation of fluorescent spectra, hydrophobic interaction was shown as the major driving force for this nanocomplex structure. CSPH-Ca-VD 3 nanocomplex possessed excellent stability during simulated gastrointestinal digestion, contributing to the prevention of acid degradation of VD 3 and the enhancement of calcium solubility. Furthermore, the calcium transport efficiency in the form of CSPH-Ca-VD 3 (4 mg/mL) across a Caco-2 cells monolayer was significantly increased 2.3-fold compared to that of free Ca 2+ , mainly attributed to the upregulation in the presence of CSPH-Ca-VD 3 of TRPV6, calbindin D9k and PMCA1b expression in Caco-2 cells. The present study provided a basis for developing a novel delivery system of peptides-calcium chelate with the dual effects of VD 3 protection and calcium uptake promotion.
研究领域:
材料科学
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纳米技术
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生物医学工程
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药学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
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