MOLECULAR BIOTECHNOLOGY
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2023
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IF 2.6
Liu Hu,Li Tao,Ma Ben,Wang Yue,Sun Jun
Osteoblast regeneration, characterized by osteoblast differentiation, is the basis of fracture healing and accelerates fracture repair. It has been reported that hyaluronan and proteoglycan link protein 1 (HAPLN1) is overexpressed during osteoblast differentiation and regulates cartilage regeneration, but its function in fracture healing remains unclear. To elucidate this issue, we collected clinical blood samples of fracture healing, established a femoral fracture rat model, and induced an osteoblast differentiation cell model. We found that HAPLN1 was overexpressed in the serum of patients with fracture healing and the bone tissues of rats with fracture healing. Furthermore, the expression of HAPLN1 was increased time dependently during the osteogenic differentiation of MC3T3-E1 cells. HAPLN1 silencing prevented osteoblast differentiation and mineralization in MC3T3-E1 cells as evidenced by decreased osteoblast differentiation-related factors, suppressed alkaline phosphatase activities, and reduced alizarin red positive staining. Mechanically, the bone morphogenic protein 4 (BMP4)/Smad1/5/8 pathway, a facilitator of osteoblastic differentiation, was found to be inhibited by HAPLN1 knockdown, and inhibition of BMP4/Smad1/5/8 signaling enhanced the effects caused by HAPLN1 silencing. These findings demonstrated that HAPLN1 might promote fracture healing by facilitating osteogenic differentiation through the BMP4/Smad1/5/8 pathway, indicating that targeting HAPLN1 may be a feasible therapeutic candidate for fracture repair. Graphical Abstract
研究领域:
骨科
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
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2020
·Vol. 24
·期 21
·12476-12490
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IF 4.486
Zhongping Su,Pengyu Zong,Ji Chen,Shuo Yang,Yihui Shen,Yan Lu,Chuanxi Yang,Xiangqing Kong,Yanhui Sheng,Wei Sun
Vascular calcification is an important risk factor for the mortality and morbidity in chronic kidney disease (CKD). Unfortunately, until now there is no certain medication targeting vascular calcification in CKD. In this study, we explored the inhibitory effect of celastrol on high calcium–induced vascular calcification and the underlying molecular mechanisms. Cell proliferation assay showed that celastrol inhibited aortic valve interstitial cell (VIC) and vascular smooth muscle cell (VSMC) proliferation when its concentration was higher than 0.6 μmol/L. 0.8 μmol/L celastrol inhibited the expression of osteogenic genes and calcium deposition induced by high-calcium medium in both AVICs and VSMCs. In mouse vascular calcification model induced by adenine combined with vitamin D, alizarin red and immunostaining showed that celastrol inhibited pro-calcification gene expression and calcium deposition in aortic wall and aortic valve tissues. At the molecular level, celastrol inhibited the increase of BMP2, phosphorylated Smad1/5 (p-Smad1/5) and non-phosphorylated β-catenin (n-p-β-catenin) induced by high-calcium medium both in vitro and in vivo. Also, BMP2 overexpression reversed the anti-calcification effects of celastrol by recovering the decrease of p-Smad1/5 and n-p-β-catenin. Furthermore, celastrol prevented the up-regulation of BMPRII and down-regulation of Smad6 induced by high calcium, and this protectory effect can be abolished by BMP2 overexpression. In conclusion, our data for the first time demonstrate that celastrol attenuates high calcium–induced arterial and valvular calcification by inhibiting BMP2/Smad1/5 signalling, which may provide a novel therapeutic strategy for arterial and valvular calcification in patients with CKD.
研究领域:
药理学
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心血管疾病
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肾病学
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分子生物学
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DOI
PubMed
高影响因子
ACTA PHARMACOLOGICA SINICA
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2026
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IF 10.4
Zhou Yan,Yang Yifan,Wang Yuehan,Cheang Wai San
YAP/TAZ and SMAD1/5 signaling modulate atherosclerosis. These involvements in diabetes-associated endothelial dysfunction remain unexplored, thereby being investigated from different endothelial cells and a total of 102 mice in the current study. Male C57BL/6 mice were fed a high-fat diet (45% kcal% fat, 15 weeks) and orally administered with endoplasmic reticulum (ER) stress alleviator 4-phenylbutyric acid (100 mg·kg –1 ·d –1 ) and jatrorrhizine (50 mg·kg –1 ·d –1 ) for 5 weeks. Endothelial cells were cultured with high glucose, ER stress inducer tunicamycin, ER stress alleviators, YAP inhibitor simvastatin, or jatrorrhizine. The mouse aorta was divided into the aortic arch and thoracic aorta regions to investigate the regional difference. Treatment with PBA and jatrorrhizine ameliorated endothelium-dependent relaxations as well as inhibited ER stress, YAP/TAZ-SMAD1/5 signaling and oxidative stress in aortas. Phosphorylation of YAP/TAZ was highly expressed in the thoracic aorta but low in the aortic arch. On the other hand, phosphorylation of SMAD1/5 was upregulated in the inner curvature of the aortic arch but downregulated in the thoracic aorta. In HUVECs, ER stress alleviators significantly inhibited YAP/TAZ-SMAD1/5 signaling and increased NO bioavailability, but the YAP inhibitor did not suppress ER stress. Inhibition of YAP/TAZ downregulated SMAD1/5 signaling under high glucose stimulation, followed by the Akt/eNOS pathway. YAP/TAZ directly interacts with SMAD1/5, and their binding is significantly enhanced under high-glucose conditions. A natural compound, jatrorrhizine, was identified to inhibit YAP/TAZ-SMAD1/5 signaling and protect vascular function in diabetes. To conclude, ER stress activates YAP/TAZ and SMAD1/5 signaling, resulting in endothelial dysfunction in diabetes. Inhibition of YAP/TAZ-SMAD1/5, such as by ER stress alleviators and natural product jatrorrhizine, ameliorates endothelial dysfunction associated with diabetes. These findings support the cross-talk between ER stress and YAP/TAZ-SMAD1/5 as well as their potential as therapeutic targets.
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DOI
PubMed
高影响因子
CytoJournal
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2025
·
IF 3.1
Xunjia Li,Zhixin Xu,Ying Li,Yan Luo,Jie Zhou,Deyu Zuo,Weijian Xiong
Objective:
Patients with chronic kidney disease (CKD) exhibit increased vascular calcification (VC) risks, worsened by high-dose erythropoietin (EPO). While EPO treats anemia, its role in VC pathogenesis remains unclear. Ginsenoside Rb1 (Rb1), a Panax ginseng compound with anti-calcification properties, may counteract EPO-induced VC through the GATA binding protein 6 (GATA6)/bone morphogenetic protein 2 (BMP2)/Smad1/5/9 pathway. This article aims to explore whether Rb1 could counteract EPO-induced VC through the GATA6/BMP2/Smad1/5/9 pathway.
Material and Methods:
Adenine-induced CKD rats and b-glycerophosphate-treated vascular smooth muscle cells (VSMCs) received EPO ± Rb1. Calcification was assessed through von Kossa/alizarin red staining. Smooth muscle protein 22-a (SM22a)/a-Smooth muscle actin (a-SMA) expression was measured by immunofluorescence and real-time-quantitative polymerase chain reaction (RT-qPCR). GATA6/BMP2/Smad1/5/9 activation was analyzed using RT-qPCR/Western blot. Rb1-BMP2 interactions were tested through biotin pulldown, micro-thermophoresis, and Co-immunoprecipitation (Co-IP). GATA6 knockdown validated pathway roles.
Results:
High-dose EPO significantly worsened CKD-associated calcification and VSMC calcification (P < 0.01), suppressed SM22a and a-SMA expression levels, and activated the GATA6/BMP2/Smad1/5/9 pathway (P < 0.01). GATA6 knockdown reduced EPO-exacerbated calcification and modulated BMP2/Smad1/5/9 signaling (P < 0.01). Rb1 increased SM22a and a-SMA expression levels and inhibited Smad 1/5/9 phosphorylation (P < 0.01), without affecting GATA6 or BMP2 expression (P > 0.05). Molecular docking and Co-IP experiments revealed that Rb1 binds directly to BMP2, blocking its interaction with bone morphogenetic protein receptor and inhibiting Smad 1/5/9 phosphorylation (P < 0.01).
Conclusion:
Rb1 mitigates EPO-aggravated VC in CKD by disrupting BMP2/Smad1/5/9 signaling, positioning it as a promising molecular intervention strategy to reduce EPO-induced vascular toxicity.
Keywords: Bone morphogenetic protein 2/Smad1/5/9 signaling pathway, Chronic kidney disease, Erythropoietin, Ginsenoside Rb1, Vascular calcification
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PubMed
高影响因子
JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION
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2021
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IF 3.517
Caiyun Liu,Zhenzu Bai,Jiayu Lin,Kai Jiang,Shan Huang,Weijia Zheng,Ruifen Chen,Yang Xiang,Xiaoyan Wang,Long Liu
The application of 3 D printing technology in tissue engineering has become increasingly important. However, due to the limitations of bio-ink, there are still some remaining problems. For example, the major challenge for ideal bio-ink is to maintain stable 3 D structure and good biocompatibility in the meantime while conventional gels are week and nearly unprintable. So, the development of new bio-ink material with improved rheological and mechanical properties is highly demanded to avoid compromising biocompatibility for tissue engineering. Silk fibroin (SF), a natural degradable polymer, is considered to be a proper material for the preparation of bio-inks. We used SF, gelatin, and polyols as raw materials to fabricate bio-inks and scaffolds. We evaluated the rheological properties and printability of bio-inks with a rotational rheometer and a 3 D printer. The scaffolds were prepared by crosslinking and freeze-drying technologies. The biocompatibility and osteoinductive functions of scaffolds were investigated by evaluating proliferation, osteogenic differentiation and related cell signaling of cultured MC3T3-E1 cells. The results showed that the scaffolds using SF, Gel and propanediol (PG) not only had good rheological properties and storage modulus, but also could better enhance osteogenic specific genes expression mediated by Smad1/5/8 and Runx2 pathways. What is more, morphological characterization showed that α-mem incubation could help scaffold form porous structure on its surface, which could shed a light on a new 3 D bio-printed bone repair scaffold with both naturally emerged and CAD-designed porous structure. Our findings provide a potential biomaterial for the treatment of bone tissue regeneration.
研究领域:
生物医学工程
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材料科学
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组织工程
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生物材料
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DOI
PubMed
高影响因子
MATERIALS & DESIGN
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2023
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IF 8.4
Shengqi Zang,Can Xiao,Maodian He,Bo Chen,Bingyao Liu,Shuai Yi,Lei Wang,Junxia Li,Xiaolei Shi,Shoushan Bu,Lei Jin
Biomimetic surface modification of scaffolds with osteoinductive molecules is a promising strategy for improving the bioactivity of scaffold and stimulating stem cell signals in bone tissue engineering. Zirconia (ZrO 2 ) offers excellent mechanical properties and biocompatibility, but its bio-inert nature hinders its use in bone regeneration applications. Recombinant human bone morphogenetic protein 7 (rhBMP-7) and arginine-glycine-aspartate (RGD) peptide were investigated for their potential use as surface-modifying biomolecules on the ZrO 2 scaffold. Our results showed that human dental pulp stem cells (hDPSCs) were induced to osteogenic differentiation by rhBMP-7 and RGD in a dose-dependent manner. Significantly enhanced alkaline phosphatase activity and up-regulated expression of osteogenic genes in hDPSCs were associated with rhBMP-7 and RGD grafting onto the ZrO 2 scaffold. Western blot analysis revealed that rhBMP-7 and RGD grafting led to activation of the SMAD1/5, p38 MARK, and ERK signaling pathways during hDPSCs differentiation. After 4 and 8 weeks of transplantation, the hDPSCs-seeded ZrO 2 -RGD-BMP-7 scaffold facilitated osteogenic differentiation and enhanced in vivo bone formation in critical-sized calvarial bone defects. The results support the simultaneous use of rhBMP-7 and RGD as surface-modifying biomolecules and hDPSCs as a source of osteogenic stem cells in conjugation with ZrO 2 -based porous scaffold for bone tissue engineering.
研究领域:
组织工程
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生物材料
·
干细胞生物学
·
骨再生
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）