Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species

Kai Chen, Pengcheng Qiu, Yu Yuan, Lin Zheng, Jianbo He, Chao Wang, Qiang Guo, Jacob Kenny, Qian Liu, Jinmin Zhao, Junhao Chen, Jennifer Tickner, Shunwu Fan, Xianfeng Lin, Jiake Xu

Research output: Contribution to journalArticle

2 Citations (Scopus)
33 Downloads (Pure)

Abstract

Rationale: Growing evidence indicates that intracellular reactive oxygen species (ROS) accumulation is a critical factor in the development of osteoporosis by triggering osteoclast formation and function. Pseurotin A (Pse) is a secondary metabolite isolated from Aspergillus fumigatus with antioxidant properties, recently shown to exhibit with a wide range of potential therapeutic applications. However, its effects on osteoporosis remain unknown. This study aimed
to explore whether Pse, by suppressing ROS levels, is able to inhibit osteoclastogenesis and prevent the bone loss induced by estrogen-deficiency in ovariectomized (OVX) mice.
Methods: The effects of Pse on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclastogenesis and bone resorptive function were examined by tartrate resistant acid phosphatase (TRAcP) staining and hydroxyapatite resorption assay. 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) was used to detect intracellular ROS production in vitro. Western blot assay was used to identify proteins associated with ROS generation and scavenging as well as ROS-mediated signaling cascades including mitogenactivated protein kinases (MAPKs), NF-κB pathways, and nuclear factor of activated T cells 1(NFATc1) activation. The expression of osteoclast-specific genes was assessed by qPCR. The in vivo potential of Pse was determined using an OVX mouse model administered with Pse or vehicle for 6 weeks. In vivo ROS production was assessed by intravenous injection of dihydroethidium (DHE) into OVX mice 24h prior to killing. After sacrifice, the bone samples were analyzed using μCT and histomorphometry to determine bone volume, osteoclast activity,
and ROS level in vivo.
Results: Pse was demonstrated to inhibit osteoclastogenesis and bone resorptive function in vitro, as well as the downregulation of osteoclast-specific genes including Acp5 (encoding TRAcP), Ctsk (encoding cathepsin K), and Mmp9 (encoding matrix metalloproteinase 9). Mechanistically, Pse suppressed intracellular ROS level by inhibiting RANKL-induced ROS production and enhancing ROS scavenging enzymes, subsequently suppressing MAPK pathway
activation (ERK, P38, and JNK) and NF-κB pathways, as well as inhibiting NFATc1 signaling. μCT and histological results indicated that OVX resulted in significant bone loss, with dramatically increased numbers of osteoclasts on the bone surface as well as increased ROS levels in the bone marrow microenvironment; whereas Pse supplementation was capable of effectively preventing these OVX-induced changes.
Conclusion: Pse was demonstrated for the first time as a novel alternative therapy for osteoclastrelated bone disease such as osteoporosis through suppressing ROS levels.
Original languageEnglish
Pages (from-to)1634-1650
JournalTheranostics
Volume9
Issue number6
DOIs
Publication statusPublished - 28 Feb 2019

Fingerprint

Osteogenesis
Reactive Oxygen Species
Bone and Bones
Osteoclasts
Osteoporosis
NFATC Transcription Factors
Protein Kinases
pseurotin
Cathepsin K
Aspergillus fumigatus
Bone Diseases
Matrix Metalloproteinase 9
Durapatite
Cytoplasmic and Nuclear Receptors
Complementary Therapies
Intravenous Injections
Genes
Estrogens
Down-Regulation
Antioxidants

Cite this

Chen, Kai ; Qiu, Pengcheng ; Yuan, Yu ; Zheng, Lin ; He, Jianbo ; Wang, Chao ; Guo, Qiang ; Kenny, Jacob ; Liu, Qian ; Zhao, Jinmin ; Chen, Junhao ; Tickner, Jennifer ; Fan, Shunwu ; Lin, Xianfeng ; Xu, Jiake. / Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species. In: Theranostics. 2019 ; Vol. 9, No. 6. pp. 1634-1650.
@article{3ce26d0373bd4f32a5809e13fcf1dfa4,
title = "Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species",
abstract = "Rationale: Growing evidence indicates that intracellular reactive oxygen species (ROS) accumulation is a critical factor in the development of osteoporosis by triggering osteoclast formation and function. Pseurotin A (Pse) is a secondary metabolite isolated from Aspergillus fumigatus with antioxidant properties, recently shown to exhibit with a wide range of potential therapeutic applications. However, its effects on osteoporosis remain unknown. This study aimedto explore whether Pse, by suppressing ROS levels, is able to inhibit osteoclastogenesis and prevent the bone loss induced by estrogen-deficiency in ovariectomized (OVX) mice.Methods: The effects of Pse on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclastogenesis and bone resorptive function were examined by tartrate resistant acid phosphatase (TRAcP) staining and hydroxyapatite resorption assay. 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) was used to detect intracellular ROS production in vitro. Western blot assay was used to identify proteins associated with ROS generation and scavenging as well as ROS-mediated signaling cascades including mitogenactivated protein kinases (MAPKs), NF-κB pathways, and nuclear factor of activated T cells 1(NFATc1) activation. The expression of osteoclast-specific genes was assessed by qPCR. The in vivo potential of Pse was determined using an OVX mouse model administered with Pse or vehicle for 6 weeks. In vivo ROS production was assessed by intravenous injection of dihydroethidium (DHE) into OVX mice 24h prior to killing. After sacrifice, the bone samples were analyzed using μCT and histomorphometry to determine bone volume, osteoclast activity,and ROS level in vivo.Results: Pse was demonstrated to inhibit osteoclastogenesis and bone resorptive function in vitro, as well as the downregulation of osteoclast-specific genes including Acp5 (encoding TRAcP), Ctsk (encoding cathepsin K), and Mmp9 (encoding matrix metalloproteinase 9). Mechanistically, Pse suppressed intracellular ROS level by inhibiting RANKL-induced ROS production and enhancing ROS scavenging enzymes, subsequently suppressing MAPK pathwayactivation (ERK, P38, and JNK) and NF-κB pathways, as well as inhibiting NFATc1 signaling. μCT and histological results indicated that OVX resulted in significant bone loss, with dramatically increased numbers of osteoclasts on the bone surface as well as increased ROS levels in the bone marrow microenvironment; whereas Pse supplementation was capable of effectively preventing these OVX-induced changes.Conclusion: Pse was demonstrated for the first time as a novel alternative therapy for osteoclastrelated bone disease such as osteoporosis through suppressing ROS levels.",
author = "Kai Chen and Pengcheng Qiu and Yu Yuan and Lin Zheng and Jianbo He and Chao Wang and Qiang Guo and Jacob Kenny and Qian Liu and Jinmin Zhao and Junhao Chen and Jennifer Tickner and Shunwu Fan and Xianfeng Lin and Jiake Xu",
year = "2019",
month = "2",
day = "28",
doi = "10.7150/thno.30206",
language = "English",
volume = "9",
pages = "1634--1650",
journal = "Theranostics",
issn = "1838-7640",
publisher = "Ivyspring International Publisher",
number = "6",

}

Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species. / Chen, Kai; Qiu, Pengcheng; Yuan, Yu; Zheng, Lin; He, Jianbo; Wang, Chao; Guo, Qiang ; Kenny, Jacob; Liu, Qian ; Zhao, Jinmin; Chen, Junhao; Tickner, Jennifer; Fan, Shunwu; Lin, Xianfeng; Xu, Jiake.

In: Theranostics, Vol. 9, No. 6, 28.02.2019, p. 1634-1650.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species

AU - Chen, Kai

AU - Qiu, Pengcheng

AU - Yuan, Yu

AU - Zheng, Lin

AU - He, Jianbo

AU - Wang, Chao

AU - Guo, Qiang

AU - Kenny, Jacob

AU - Liu, Qian

AU - Zhao, Jinmin

AU - Chen, Junhao

AU - Tickner, Jennifer

AU - Fan, Shunwu

AU - Lin, Xianfeng

AU - Xu, Jiake

PY - 2019/2/28

Y1 - 2019/2/28

N2 - Rationale: Growing evidence indicates that intracellular reactive oxygen species (ROS) accumulation is a critical factor in the development of osteoporosis by triggering osteoclast formation and function. Pseurotin A (Pse) is a secondary metabolite isolated from Aspergillus fumigatus with antioxidant properties, recently shown to exhibit with a wide range of potential therapeutic applications. However, its effects on osteoporosis remain unknown. This study aimedto explore whether Pse, by suppressing ROS levels, is able to inhibit osteoclastogenesis and prevent the bone loss induced by estrogen-deficiency in ovariectomized (OVX) mice.Methods: The effects of Pse on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclastogenesis and bone resorptive function were examined by tartrate resistant acid phosphatase (TRAcP) staining and hydroxyapatite resorption assay. 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) was used to detect intracellular ROS production in vitro. Western blot assay was used to identify proteins associated with ROS generation and scavenging as well as ROS-mediated signaling cascades including mitogenactivated protein kinases (MAPKs), NF-κB pathways, and nuclear factor of activated T cells 1(NFATc1) activation. The expression of osteoclast-specific genes was assessed by qPCR. The in vivo potential of Pse was determined using an OVX mouse model administered with Pse or vehicle for 6 weeks. In vivo ROS production was assessed by intravenous injection of dihydroethidium (DHE) into OVX mice 24h prior to killing. After sacrifice, the bone samples were analyzed using μCT and histomorphometry to determine bone volume, osteoclast activity,and ROS level in vivo.Results: Pse was demonstrated to inhibit osteoclastogenesis and bone resorptive function in vitro, as well as the downregulation of osteoclast-specific genes including Acp5 (encoding TRAcP), Ctsk (encoding cathepsin K), and Mmp9 (encoding matrix metalloproteinase 9). Mechanistically, Pse suppressed intracellular ROS level by inhibiting RANKL-induced ROS production and enhancing ROS scavenging enzymes, subsequently suppressing MAPK pathwayactivation (ERK, P38, and JNK) and NF-κB pathways, as well as inhibiting NFATc1 signaling. μCT and histological results indicated that OVX resulted in significant bone loss, with dramatically increased numbers of osteoclasts on the bone surface as well as increased ROS levels in the bone marrow microenvironment; whereas Pse supplementation was capable of effectively preventing these OVX-induced changes.Conclusion: Pse was demonstrated for the first time as a novel alternative therapy for osteoclastrelated bone disease such as osteoporosis through suppressing ROS levels.

AB - Rationale: Growing evidence indicates that intracellular reactive oxygen species (ROS) accumulation is a critical factor in the development of osteoporosis by triggering osteoclast formation and function. Pseurotin A (Pse) is a secondary metabolite isolated from Aspergillus fumigatus with antioxidant properties, recently shown to exhibit with a wide range of potential therapeutic applications. However, its effects on osteoporosis remain unknown. This study aimedto explore whether Pse, by suppressing ROS levels, is able to inhibit osteoclastogenesis and prevent the bone loss induced by estrogen-deficiency in ovariectomized (OVX) mice.Methods: The effects of Pse on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced osteoclastogenesis and bone resorptive function were examined by tartrate resistant acid phosphatase (TRAcP) staining and hydroxyapatite resorption assay. 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) was used to detect intracellular ROS production in vitro. Western blot assay was used to identify proteins associated with ROS generation and scavenging as well as ROS-mediated signaling cascades including mitogenactivated protein kinases (MAPKs), NF-κB pathways, and nuclear factor of activated T cells 1(NFATc1) activation. The expression of osteoclast-specific genes was assessed by qPCR. The in vivo potential of Pse was determined using an OVX mouse model administered with Pse or vehicle for 6 weeks. In vivo ROS production was assessed by intravenous injection of dihydroethidium (DHE) into OVX mice 24h prior to killing. After sacrifice, the bone samples were analyzed using μCT and histomorphometry to determine bone volume, osteoclast activity,and ROS level in vivo.Results: Pse was demonstrated to inhibit osteoclastogenesis and bone resorptive function in vitro, as well as the downregulation of osteoclast-specific genes including Acp5 (encoding TRAcP), Ctsk (encoding cathepsin K), and Mmp9 (encoding matrix metalloproteinase 9). Mechanistically, Pse suppressed intracellular ROS level by inhibiting RANKL-induced ROS production and enhancing ROS scavenging enzymes, subsequently suppressing MAPK pathwayactivation (ERK, P38, and JNK) and NF-κB pathways, as well as inhibiting NFATc1 signaling. μCT and histological results indicated that OVX resulted in significant bone loss, with dramatically increased numbers of osteoclasts on the bone surface as well as increased ROS levels in the bone marrow microenvironment; whereas Pse supplementation was capable of effectively preventing these OVX-induced changes.Conclusion: Pse was demonstrated for the first time as a novel alternative therapy for osteoclastrelated bone disease such as osteoporosis through suppressing ROS levels.

U2 - 10.7150/thno.30206

DO - 10.7150/thno.30206

M3 - Article

VL - 9

SP - 1634

EP - 1650

JO - Theranostics

JF - Theranostics

SN - 1838-7640

IS - 6

ER -