scholarly journals Different effects of Wnt/β-catenin activation and PTH activation in adult and aged male mice metaphyseal fracture healing

2020 ◽  
Author(s):  
Daocheng Liu ◽  
Hao Qin ◽  
Jiazhi Yang ◽  
Lei Yang ◽  
Sihao He ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Signaling Pathway ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mrna Levels ◽  
Wild Type ◽  
Long Term Effects ◽  
Femoral Metaphysis ◽  
Related Transcription Factor ◽  
Early Effects

Abstract Background: Fractures in older men are not uncommon and need to be healed as soon as possible to avoid related complications. Anti-osteoporotic drugs targeting Wnt/β-catenin and PTH (parathyroid hormone) to promote fracture healing have become an important direction in recent years. The study is to observe whether there is a difference in adult and aged situations by activating two signal paths. Methods: A single cortical hole with a diameter of 0.6 mm was made in the femoral metaphysis of Catnb lox(ex3) mice and wild-type mice. The fracture healing effects of CA (Wnt/β-catenin activation) and PTH (activated by PTH (1–34) injections) were assessed by X-ray and CT imaging on days 7, 14, and 21 after fracture. The mRNA levels of β-catenin, PTH1R( Parathyroid hormone 1 receptor ), and RUNX2(Runt-related transcription factor 2) in the fracture defect area were detected using RT-PCR. Angiogenesis and osteoblasts were observed by immunohistochemistry and osteoclasts were observed by TRAP (Tartrate-resistant Acid Phosphatase). Result: Adult CA mice and adult PTH mice showed slightly better fracture healing than adult wild-type (WT) mice, but there was no statistical difference. Aged CA mice showed better promotion of angiogenesis and osteoblasts and better fracture healing than aged PTH mice. Conclusion: The application of Wnt/β-catenin signaling pathway drugs for fracture healing in elderly patients may bring better early effects than PTH signaling pathway drugs, but the long-term effects need to be observed.

scholarly journals Different effects of Wnt/β-catenin activation and PTH activation in adult and aged male mice metaphyseal fracture healing

2020 ◽  
Author(s):  
Daocheng Liu ◽  
Hao Qin ◽  
Jiazhi Yang ◽  
Lei Yang ◽  
Sihao He ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Signaling Pathway ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mrna Levels ◽  
Wild Type ◽  
Long Term Effects ◽  
Femoral Metaphysis ◽  
Related Transcription Factor ◽  
Early Effects

Abstract Background: Fractures in older men are not uncommon and need to be healed as soon as possible to avoid related complications. Anti-osteoporotic drugs targeting Wnt/β-catenin and PTH (parathyroid hormone) to promote fracture healing have become an important direction in recent years.Objective: Observe whether there is a difference in adult and aged situations by activating two signal paths.Methods: A single cortical hole with a diameter of 0.6 mm was made in the femoral metaphysis of Catnblox(ex3) mice and wild-type mice. The fracture healing effects of CA(Wnt/β-catenin activation) and PTH (activated by PTH (1–34) injections) were assessed by X-ray and CT imaging on days 7, 14, and 21 after fracture. The mRNA levels of β-catenin, PTH1R(Parathyroid hormone 1 receptor), and RUNX2(Runt-related transcription factor 2) in the fracture defect area were detected using RT-PCR. Angiogenesis and osteoblasts were observed by immunohistochemistry and osteoclasts were observed by TRAP (Tartrate-resistant Acid Phosphatase).Result: Adult CA mice and adult PTH mice showed slightly better fracture healing than adult wild-type (WT) mice, but there was no statistical difference. Aged CA mice showed better promotion of angiogenesis and osteoblasts and better fracture healing than aged PTH mice.Conclusion: The application of Wnt/β-catenin signaling pathway drugs for fracture healing in elderly patients may bring better early effects than PTH signaling pathway drugs, but the long-term effects need to be observed.

scholarly journals Different effects of Wnt/β-catenin activation and PTH activation in adult and aged male mice metaphyseal fracture healing

2019 ◽  
Author(s):  
Daocheng Liu ◽  
Hao Qin ◽  
Jiazhi Yang ◽  
Lei Yang ◽  
Sihao He ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Signaling Pathway ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mrna Levels ◽  
Wild Type ◽  
Long Term Effects ◽  
Femoral Metaphysis ◽  
Related Transcription Factor ◽  
Early Effects

Abstract Background: Fractures in older men are not uncommon and need to be healed as soon as possible to avoid related complications. Anti-osteoporotic drugs targeting Wnt/β-catenin and PTH (parathyroid hormone) to promote fracture healing have become an important direction in recent years. Objective: Observe whether there is a difference in adult and aged situations by activating two signal paths. Methods: A single cortical hole with a diameter of 0.6 mm was made in the femoral metaphysis of Catnblox(ex3) mice and wild-type mice. The fracture healing effects of CA(Wnt/β-catenin activation) and PTH (activated by tamoxifen and PTH (1–34) injections) were assessed by X-ray and CT imaging on days 7, 14, and 21 after fracture. The mRNA levels of β-catenin, PTH1R(Parathyroid hormone 1 receptor), and RUNX2(Runt-related transcription factor 2) in the fracture defect area were detected using RT-PCR. Angiogenesis and osteoblasts were observed by immunohistochemistry and osteoclasts were observed by TRAP (Tartrate-resistant Acid Phosphatase). Result: Adult CA mice and adult PTH mice showed slightly better fracture healing than adult wild-type (WT) mice, but there was no statistical difference. Aged CA mice showed better promotion of angiogenesis and osteoblasts and better fracture healing than aged PTH mice. Conclusion: The application of Wnt/β-catenin signaling pathway drugs for fracture healing in elderly patients may bring better early effects than PTH signaling pathway drugs, but the long-term effects need to be observed.

scholarly journals Different effects of Wnt/β-catenin activation and PTH activation in adult and aged male mice metaphyseal fracture healing

2020 ◽  
Author(s):  
Daocheng Liu ◽  
Hao Qin ◽  
Jiazhi Yang ◽  
Lei Yang ◽  
Sihao He ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Signaling Pathway ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mrna Levels ◽  
Wild Type ◽  
Long Term Effects ◽  
Femoral Metaphysis ◽  
Related Transcription Factor ◽  
Early Effects

Abstract Background: Fractures in older men are not uncommon and need to be healed as soon as possible to avoid related complications. Anti-osteoporotic drugs targeting Wnt/β-catenin and PTH (parathyroid hormone) to promote fracture healing have become an important direction in recent years. Objective: Observe whether there is a difference in adult and aged situations by activating two signal paths. Methods: A single cortical hole with a diameter of 0.6 mm was made in the femoral metaphysis of Catnblox(ex3) mice and wild-type mice. The fracture healing effects of CA(Wnt/β-catenin activation) and PTH (activated by PTH (1–34) injections) were assessed by X-ray and CT imaging on days 7, 14, and 21 after fracture. The mRNA levels of β-catenin, PTH1R(Parathyroid hormone 1 receptor), and RUNX2(Runt-related transcription factor 2) in the fracture defect area were detected using RT-PCR. Angiogenesis and osteoblasts were observed by immunohistochemistry and osteoclasts were observed by TRAP (Tartrate-resistant Acid Phosphatase). Result: Adult CA mice and adult PTH mice showed slightly better fracture healing than adult wild-type (WT) mice, but there was no statistical difference. Aged CA mice showed better promotion of angiogenesis and osteoblasts and better fracture healing than aged PTH mice. Conclusion: The application of Wnt/β-catenin signaling pathway drugs for fracture healing in elderly patients may bring better early effects than PTH signaling pathway drugs, but the long-term effects need to be observed.

scholarly journals Double-strand break repair deficiency in NONO knockout murine embryonic fibroblasts and compensation by spontaneous upregulation of the PSPC1 paralog

2014 ◽  
Vol 42 (15) ◽  
pp. 9771-9780 ◽  
Author(s):  
Shuyi Li ◽  
Zhentian Li ◽  
Feng-Jue Shu ◽  
Hairong Xiong ◽  
Andrew C. Phillips ◽  
...  
Keyword(s):  
Genotoxic Stress ◽  
Strand Break ◽  
Double Strand Break ◽  
Stable Complex ◽  
Mrna Levels ◽  
Wild Type ◽  
Long Term Effects ◽  
Dsb Repair ◽  
Embryonic Fibroblasts ◽  
Compensatory Response

Abstract NONO, SFPQ and PSPC1 make up a family of proteins with diverse roles in transcription, RNA processing and DNA double-strand break (DSB) repair. To understand long-term effects of loss of NONO, we characterized murine embryonic fibroblasts (MEFs) from knockout mice. In the absence of genotoxic stress, wild-type and mutant MEFs showed similar growth rates and cell cycle distributions, and the mutants were only mildly radiosensitive. Further investigation showed that NONO deficiency led to upregulation of PSPC1, which replaced NONO in a stable complex with SFPQ. Knockdown of PSPC1 in a NONO-deficient background led to severe radiosensitivity and delayed resolution of DSB repair foci. The DNA-dependent protein kinase (DNA-PK) inhibitor, NU7741, sensitized wild-type and singly deficient MEFs, but had no additional effect on doubly deficient cells, suggesting that NONO/PSPC1 and DNA-PK function in the same pathway. We tested whether NONO and PSPC1 might also affect repair indirectly by influencing mRNA levels for other DSB repair genes. Of 12 genes tested, none were downregulated, and several were upregulated. Thus, NONO or related proteins are critical for DSB repair, NONO and PSPC1 are functional homologs with partially interchangeable functions and a compensatory response involving PSPC1 blunts the effect of NONO deficiency.

scholarly journals Role of Zip1 in the regulation of NPY expression by MLT to promote fracture healing in rats

2020 ◽  
Vol 64 (s2) ◽  
Author(s):  
Junjun Shang ◽  
Bin Ma ◽  
Guiling Zhu ◽  
Penghong Dong ◽  
Chan Wang ◽  
...  
Keyword(s):  
Fracture Healing ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Type I Collagen ◽  
Femoral Fractures ◽  
Nodule Formation ◽  
Type I ◽  
Y1 Receptor ◽  
Related Transcription Factor

Our previous study documented that melatonin (MLT) induced the osteogenic differentiation of mesenchymal stem cells (MSCs) and promoted the healing of femoral fractures in rats via the neuropeptide Y (NPY)/neuropeptide Y1 receptor (NPY1R) signaling pathway. MLT treatment upregulated the expression of the zinc uptake transporter zinc transporter 1 (Zip1) in nerve cells. Prior research demonstrated that oral zinc upregulated NPY expression. MSCs were isolated from rat bone marrow and identified using flow cytometry in our study. The results showed that MLT treatment upregulated NPY and NPY1R levels in MSCs with osteogenic differentiation, which was accompanied by upregulated Zip1 expression. However, the MLT-induced osteogenic differentiation of MSCs was reversed after interference of Zip1 expression. It was confirmed by the decreased alkaline phosphatase (ALP) level; downregulated activities of type I collagen α1 chain (COL1A1), osteocalcin (OCN), runt-related transcription factor 2 (Runx2) and ALP; and reduced mineralized nodule formation. MLT promoted fracture healing in rats with femoral fracture, which was accompanied by increased expression of NPY and NPY1R and significantly increased expression of Zip1. In contrast, the silencing of Zip1 expression reversed MLT-mediated fracture healing. In summary, Zip1 participated in the regulation of the NPY/NPY1R signaling pathway via MLT to promote the osteogenic differentiation of MSCs and fracture healing.

scholarly journals Icariin Treatment Enhanced the Skeletal Response to Exercise in Estrogen-Deficient Rats

2019 ◽  
Vol 16 (19) ◽  
pp. 3779
Author(s):  
Zhao ◽  
Bu ◽  
Chen
Keyword(s):  
Bone Loss ◽  
Mechanical Strain ◽  
Elevated Serum ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mrna Levels ◽  
Sprague Dawley ◽  
Ovx Rats ◽  
Phosphorylated Akt ◽  
Related Transcription Factor ◽  
The Difference

Estrogen deficiency frequently leads to a fall in estrogen receptor- (ER) numbers and then reduces the skeletal response to mechanical strain. It, however, is still unclear whether phytoestrogen administration will enhance the effects of exercise on the estrogen-deficient bone loss. This study aimed to determine the effect of Icariin treatment on the response of osteogenic formation to exercise in ovariectomized (OVX) rats. Thirty-two 3-month old female Sprague–Dawley rats were randomly allocated into four groups: (1) Sham-operated (SO); (2) OVX; (3) OVX plus exercise (EX); and (4) OVX plus exercise and Icariin (EI). After 8-week interventions, the rats were killed and samples were collected for bone morphometry, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot analyses. EI interventions showed a greater improvement for the OVX-induced bone loss and the elevated serum tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) compared with EX only. Both EX and EI interventions bettered the OVX-related reduction of BV/TV and trabecular number and thickness, and decreased the enlargement of trabecular bone separation (Tb. Sp); the improvement for BV/TV and Tb. Sp was greater in EI group. Furthermore, EX and EI treatment significantly increased the number of ALP+ cells and mineralized nodule areas compared with OVX group; the change was higher in EI group. Additionally, in comparison to OVX rats, the protein and mRNA expression of -catenin, phosphorylated-Akt (p-Akt) or Akt, ER, and Runt-related transcription factor 2 (Runx2) in osteoblasts were elevated in EX and EI intervention rats, with greater change observed in EI group. The upregulated -catenin and Akt mRNA levels in EX and EI groups was depressed by ICI182780 treatment, and the difference in -catenin and Akt mRNA levels between EX and EI groups was no longer significant. Conclusively, the combination of Icariin and exercise significantly prevent OVX-induced bone loss and increase osteoblast differentiation and the ability of mineralization compared with exercise alone; the changes might be regulated partly by ER/Akt/-catenin pathway.

scholarly journals Accelerated Cartilage Resorption by Chondroclasts during Bone Fracture Healing in Osteoprotegerin-Deficient Mice

Endocrinology ◽  
2009 ◽  
Vol 150 (11) ◽  
pp. 4823-4834 ◽  
Author(s):  
Norikazu Ota ◽  
Hironari Takaishi ◽  
Naoto Kosaki ◽  
Jiro Takito ◽  
Masaki Yoda ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Fracture Healing ◽  
Bone Fracture ◽  
Negative Regulator ◽  
Raw 264.7 Cells ◽  
Tartrate Resistant Acid Phosphatase ◽  
Wild Type ◽  
Bone Fracture Healing ◽  
Cartilage Resorption ◽  
Multinucleated Cells

Receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG), a decoy receptor of RANKL, maintain bone mass by regulating the differentiation of osteoclasts, which are bone-resorbing cells. Endochondral bone ossification and bone fracture healing involve cartilage resorption, a less well-understood process that is needed for replacement of cartilage by bone. Here we describe the role of OPG produced by chondrocytes in chondroclastogenesis. Fracture healing in OPG−/− mice showed faster union of the fractured bone, faster resorption of the cartilaginous callus, and an increased number of chondroclasts at the chondroosseous junctions compared with that in wild-type littermates. When a cultured pellet of OPG−/− chondrocytes was transplanted beneath the kidney capsule, the pellet recruited many chondroclasts. The pellet showed the ability to induce tartrate-resistant acid phosphatase-positive multinucleated cells from RAW 264.7 cells in vitro. Finally, OPG−/− chondrocytes (but not wild-type chondrocytes) cultured with spleen cells induced many tartrate-resistant acid phosphatase-positive multinucleated cells. The expression of RANKL and OPG in chondrocytes was regulated by several osteotropic factors including 1,25-dihydroxyvitamin D3, PTHrP, IL-1α, and TNF-α. Thus, local OPG produced by chondrocytes probably controls cartilage resorption as a negative regulator for chondrocyte-dependent chondroclastogenesis.

Mechanisms underlying the long-term regulation of NHE3 by parathyroid hormone

2008 ◽  
Vol 294 (5) ◽  
pp. F1232-F1237 ◽  
Author(s):  
Camila Nogueira Alves Bezerra ◽  
Adriana Castello Costa Girardi ◽  
Luciene Regina Carraro-Lacroix ◽  
Nancy Amaral Rebouças
Keyword(s):  
Parathyroid Hormone ◽  
Regulatory Region ◽  
Experimental Models ◽  
Cell Surface Expression ◽  
Epithelial Cell Line ◽  
Mrna Levels ◽  
Apical Surface ◽  
Long Term Effects ◽  
Renal Epithelial Cell

The activity of the Na+/H+ exchanger NHE3 is regulated by a number of factors including parathyroid hormone (PTH). In the current study, we used a renal epithelial cell line, the opossum kidney (OKP) cell, to elucidate the mechanisms underlying the long-term effects of PTH on NHE3 transport activity and expression. We observed that NHE3 activity was reduced 6 h after addition of PTH, and this reduction persisted almost unaltered after 24 h. The decrease in activity was associated with diminished NHE3 cell surface expression at 6, 16, and 24 h after PTH addition, total cellular NHE3 protein at 16 and 24 h, and NHE3 mRNA abundance at 24 h. The lower levels of NHE3 mRNA were associated to a small, but significant, decrease in mRNA stability. Additionally, by analyzing the rat NHE3 gene promoter activity in OKP cells, we verified that the regulatory region spanning the segment −152 to +55 was mildly reduced under the influence of PTH. This effect was completely abolished by the presence of the PKA inhibitor KT 5720. In conclusion, long-term exposure to PTH results in reduction of NHE3 mRNA levels due to a PKA-dependent inhibitory effect on the NHE3 promoter and a small reduction of mRNA half-life, and decrease in the total amount of protein which is preceded by endocytosis of the apical surface NHE3. The decreased NHE3 expression is likely to be responsible for the reduction of sodium, bicarbonate, and fluid reabsorption in the proximal tubule consistently perceived in experimental models of PTH disorders.

scholarly journals Endogenous Parathyroid Hormone Promotes Fracture Healing by Increasing Expression of BMPR2 through cAMP/PKA/CREB Pathway in Mice

2017 ◽  
Vol 42 (2) ◽  
pp. 551-563 ◽  
Author(s):  
Wei Zhou ◽  
Lipeng Yu ◽  
Jin Fan ◽  
Bowen Wan ◽  
Tao Jiang ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Bone Morphogenetic Protein ◽  
Signaling Pathway ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Male Mice ◽  
Morphogenetic Protein ◽  
Significant Difference ◽  
Creb Pathway

Background/Aims: Endogenous parathyroid hormone (PTH) plays an important role in fracture healing. This study investigated whether endogenous PTH regulates fracture healing by bone morphogenetic protein (BMP) and/or the transforming growth factor-β (TGF-β) signaling pathway. Methods: Eight-week-old wild-type (WT) and PTH-knockout (PTH KO) male mice were selected, and models of open right-femoral fracture were constructed. Fracture healing and callus characteristics of mice in the two groups were compared by X-ray, micro-computed tomography, histological, and immunohistochemical examinations. Bone marrow mesenchymal stem cells (BMMSCs) of 8-week-old WT and PTHKO male mice were obtained and induced into osteoblasts and chondrocytes. Results: We found that expression levels of Runt-related transcription factor (RUNX2), bone morphogenetic protein-receptor-type Ⅱ (BMPR2), phosphorylated Smad 1/5/8, and phosphorylated cyclic adenosine monophosphate-responsive element binding protein (CREB) in the callus of PTHKO mice were significantly decreased, whereas no significant difference in expression of SOX9, TGF-βR2,or pSMAD2/3 was observed between PTHKO and WT mice. Additionally, the activity of osteoblast alkaline phosphatase was low at 7 days post-induction, and was upregulated by addition of PTH or dibutyryl cyclic adenosine monophosphate (dbcAMP) to the cell culture. Furthermore, H89 (protein kinase A inhibitor)eliminated the simulating effects of PTH and dbcAMP, and a low concentration of cyclic adenosine monophosphate (cAMP) was observed in PTHKO mouse BMMSCs. Conclusion: These results suggested that endogenous PTH enhanced BMPR2 expression by a cAMP/PKA/CREB pathway in osteoblasts, and increased RUNX2 expression through transduction of the BMP/pSMAD1/5/8 signaling pathway.

Effects of Parathyroid Hormone on Osteoporotic Fracture Healing in Mice via Non-Phospholipases C-Dependent Protein Kinase C Signaling Pathway

2021 ◽  
Vol 11 (5) ◽  
pp. 903-911
Author(s):  
Lei Wang ◽  
Linjuan Liu ◽  
Sixin Sun ◽  
Li Xiao ◽  
Qinyi Jiang ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Fracture Healing ◽  
Signaling Pathway ◽  
Osteoporotic Fracture ◽  
Expression Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mineral Density ◽  
Pkc Inhibitor ◽  
Pkc Signaling

Objectives: This study was aimed to explore the effects of parathyroid hormone (PTH) on osteoporotic fracture healing in mice and the underlying mechanisms. Methods: Microarray analysis was conducted to analyze the gene expression level in MC3T3-E1 cells. Carboxyfluorescein succinimidyl ester (CFSE) staining and flow cytometry was adopted to analyze the proliferation and apopto-sis of MC3T3-E1 cells. qRT-PCR was used to analyze the mRNA expression level. Fluorescence resonance energy transfer (FRET) assay was conducted to detect PKC activity. The bone mineral density (BMD) and bone volume (BV)/total volume (TV) were determined via enzyme-linked immunosorbent assay (ELISA) and microscopic computed tomography (micro-CT). Results: ERK1/2 was abnormally expressed in MC3T3-E1 cells after GlylArg19hPTH (1-34) + KT5720 treatment. GlylArg19hPTH (1-34)+ KT5720 treatment promoted cell proliferation, inhibited cell apoptosis, and upregulatedthe expression of osteogenesis-related genes (ALP, OPN, Runx2 and OPG) in MC3T3-E1 cells, which were due to the activation of the non-PLC-dependent PKC signaling pathway and can be blocked by PKC inhibitor Go6983 or ERK1/2 inhibitor BVD-523. Moreover, the activity of PKC in MC3T3-E1 cells treated with GlylArg19hPTH (1-34) + KT5720 + Go6983 was alleviated by ERK1/2 inhibitor BVD-523. In vivo, specific activation of the non-PLC-dependent PKC signaling pathway increased the serum levels of APL and OPG in mice with osteoporotic fracture, which were reversed by PKC inhibitor Go6983 and ERK1/2 inhibitor BVD-523. Moreover, PKC inhibitor Go6983 and ERK1/2 inhibitor BVD-523 suppressed the elevation of BV/TV and BMD induced by specific activation of the non-PKC-dependent signaling pathway. Conclusions: Taken together, PTH stimulates osteoporotic fracture healing in mice through the non-PLC-dependent PKC signaling pathway in which ERK1/2 exerts a vital role.

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