P450 oxidoreductase expressed in rat chondrocytes modulates chondrogenesis via cholesterol- and Indian Hedgehog-dependent mechanisms

Aguilar, Alexandra; Wu, Shufang; De Luca, Francesco

doi:10.1210/en.2009-0043

Cited by 21Hedgehog Signaling Pathway StudiesOsteoarthritis Treatment and MechanismsProteoglycans and glycosaminoglycans research

P450 oxidoreductase expressed in rat chondrocytes modulates chondrogenesis via cholesterol- and Indian Hedgehog-dependent mechanisms

Aguilar, Alexandra, Wu, Shufang, De Luca, Francesco·2009·Endocrinology

DOI 10.1210/en.2009-0043

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Abstract

Cytochrome P450 oxidoreductase (POR) is the electron donor for microsomal cytochrome P450 enzymes and other non-P450 enzymes. Targeted deletion of POR expression in mice leads to a variety of embryonic defects, including bone abnormalities. In addition, POR mutations in humans are associated with impaired steroidogenesis and skeletal malformations. Yet, little is known on the mechanisms underlying the skeletal abnormalities secondary to impaired POR activity. In our study, rat chondrocytes transfected with POR-specific short interfering RNAs exhibited decreased cell proliferation and differentiation and induced apoptosis. In addition, the reduced expression of POR in chondrocytes caused decreased intracellular cholesterol content. The addition of cholesterol in the culture medium prevented the POR small interfering RNA (siRNA)-mediated effects on chondrocyte proliferation, differentiation, and apoptosis. Because cholesterol is required for normal activity of the hedgehog proteins, we evaluated the effects of POR siRNAs on the expression of Indian hedgehog (Ihh), an important regulator of chondrogenesis. POR siRNA-transfected chondrocytes exhibited reduced Ihh expression, with such effect being neutralized by cholesterol. Lastly, recombinant human/mouse Ihh prevented the POR siRNA-mediated effects on chondrocyte proliferation, differentiation, and apoptosis. Our findings suggest that the bone malformations associated with defective POR activity are due to reduced cholesterol synthesis and, in turn, reduced Ihh expression in chondrocytes.

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POR knockdown by RNA interference in rat chondrocytes decreased cell proliferation and differentiation, induced apoptosis, and reduced expression of Indian hedgehog, but these effects were reversed by providing cholesterol.
“Second, POR knockdown by RNA interference in rat chondrocytes decreased cell proliferation and differentiation, induced apoptosis, and reduced expression of Indian hedgehog, but these effects were reversed by providing cholesterol (343).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [343]

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Authors: Aguilar, A., Wu, S., & De Luca, F.
Affiliations: St. Christopher's Hospital for Children (US)
ORCID: Wu, Shufang
Venue: Endocrinology
Publisher: The Endocrine Society
Published: 2009-06
Volume: 150
Issue: 6
Pages: 2732-2739
ISSN: 0013-7227 (ISSN-L)
1945-7170
DOI: 10.1210/en.2009-0043
OpenAlex: W2052155743
Semantic Scholar: 521881ecb666c4e145ad99b006d0bed5eba5cdaa
Cited by: 21
References: 47
Citations by year: 2025: 1 · 2023: 3 · 2022: 1 · 2021: 2 · 2019: 1 · 2018: 1 · 2016: 1 · 2013: 3 · 2012: 1
FWCI: 1.07
Type: journal article
Language: en
Open access: closed
Locations: Endocrinology (publishedVersion)
PubMed (publishedVersion)
Funders: Saint Christopher's Hospital for Children
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Hedgehog Signaling Pathway Studies
Health Sciences › Medicine › Rheumatology › Osteoarthritis Treatment and Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
Keywords: cytochrome P450 oxidoreductase, chondrocytes, chondrogenesis, Indian hedgehog, cholesterol, Hedgehog signaling, chondrocyte proliferation, chondrogenic differentiation, apoptosis, bone malformations, steroidogenesis, siRNA knockdown
MeSH terms: Animals, Cell Differentiation, Cells, Cultured, Cholesterol, Humans, NADPH-Ferrihemoprotein Reductase, Recombinant Proteins, RNA, Messenger, Transfection, Signal Transduction, Rats, Sprague-Dawley, Apoptosis, Chondrocytes, Chondrogenesis, RNA, Small Interfering, Cell Proliferation, Mice, Rats, Hedgehog Proteins

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P450 oxidoreductase expressed in rat chondrocytes modulates chondrogenesis via cholesterol- and Indian Hedgehog-dependent mechanisms

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