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Decreased histone deacetylase 4 is associated with human osteoarthritis cartilage degeneration by releasing histone deacetylase 4 inhibition of runt-related transcription factor-2 and increasing osteoarthritis-related genes: a novel mechanism of human osteoarthritis cartilage degeneration.

Cao K, Wei L, Zhang Z, Guo L, Zhang C, Li Y, Sun C, Sun X, Wang S, Li P, Wei X - Arthritis Res. Ther. (2014)

Bottom Line: The levels of HDAC4 in the cartilage from OA patients and healthy 40- to 60-year-old donors were decreased to 31% and 65% compared with specimens from 20- to 40-year-old healthy donors, respectively (P <0.05).In addition, the data also shows that overexpression of HDAC4 not only decreased the expression of interleukin (IL)-1β, Cox2 and iNos and increased the expression of aggrecan, but also partially blocked the effect of IL-1β on expression of catabolic events in human OA chondrocytes.In contrast, inhibition of HDAC4 by TSA drug had an opposite effect.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: To investigate if decreased histone deacetylase 4 (HDAC4) is associated with human osteoarthritis (OA) cartilage degeneration by releasing HDAC4 inhibition of runt-related transcription factor-2 (Runx2) resulting in increase of OA cartilage degeneration-related genes.

Methods: The mRNA and protein levels of HDAC4, Runx2, matrix metalloproteinase (MMP)-13, Indian hedgehog (Ihh) and type X collagen were detected by performing real-time PCR (RT-PCR), western blotting and immunohistochemistry on specimens from human OA and normal cartilage. To further explore the mechanism of regulation of Runx2 and OA-related genes by HDAC4, changes in these OA-related genes were further quantified by RT-PCR after overexpression of HDAC4 and knockdown of HDAC4 by siRNA. Runx2 and MMP-13 promoter activities were measured by dual luciferase assays.

Results: The levels of HDAC4 in the cartilage from OA patients and healthy 40- to 60-year-old donors were decreased to 31% and 65% compared with specimens from 20- to 40-year-old healthy donors, respectively (P <0.05). Decreased HDAC4 was associated with increased Runx2 and other OA-related genes in human OA cartilage, specifically: MMP-13, Ihh and type X collagen. Exogenous HDAC4 decreased the mRNA levels of Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, ADAMTS-4 and -5, and increased the mRNA of type II collagen. In addition, the data also shows that overexpression of HDAC4 not only decreased the expression of interleukin (IL)-1β, Cox2 and iNos and increased the expression of aggrecan, but also partially blocked the effect of IL-1β on expression of catabolic events in human OA chondrocytes. HDAC4 also inhibited Runx2 promoter activity and MMP13 promotor activity in a dose-dependent manner. In contrast, inhibition of HDAC4 by TSA drug had an opposite effect.

Conclusions: Our study is the first to demonstrate that decreased HDAC4 contributes, at least in part, to the pathogenesis of OA cartilage degeneration. Thus, HDAC4 may have chondroprotective properties by inhibiting Runx2 and OA-related genes.

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Related in: MedlinePlus

HDAC4 downregulates OA-related gene expressions in human OA chondrocytes. Human OA chondrocytes were transfected with HDAC4-flag or HDAC4 siRNA. All transfections were performed in triplicate. Data are presented as means ± SD. *P <0.05. (A) The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 or control vector transfection. (B)The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 siRNA or scrambled siRNA control transfection. HDAC4, histone deacetylase 4; MMP, matrix metalloprotease; OA, osteoarthritis; Runx2, runt-related transcription factor-2; SD, standard deviation; siRNA, small interfering RNA; TSA, trichostatin A.
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Fig5: HDAC4 downregulates OA-related gene expressions in human OA chondrocytes. Human OA chondrocytes were transfected with HDAC4-flag or HDAC4 siRNA. All transfections were performed in triplicate. Data are presented as means ± SD. *P <0.05. (A) The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 or control vector transfection. (B)The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 siRNA or scrambled siRNA control transfection. HDAC4, histone deacetylase 4; MMP, matrix metalloprotease; OA, osteoarthritis; Runx2, runt-related transcription factor-2; SD, standard deviation; siRNA, small interfering RNA; TSA, trichostatin A.

Mentions: Our data showed that overexpression of HDAC4 decreased the mRNA levels of Ihh, type X collagen, MMP1, MMP3, MMP-13, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 in OA chondrocytes to 11%, 23%, 33.8%, 23.7%, 33%, 60%, 57.7%, 54.7%, 45.1% and 48.9%, respectively, of those in controls (Figure 5A). In addition, the mRNA levels of type II collagen and aggrecan significantly increased 4.7- and 9.0-fold after HDAC4 transfection (Figure 5A) (P <0.05). In contrast, knockdown HDAC4 by HDAC4 siRNA showed an opposite effect (Figure 5B) (P <0.05). The transfection efficiencies of HDAC4 and HDAC4 siRNA were validated by real-time PCR (Figure 5A, B).Figure 5


Decreased histone deacetylase 4 is associated with human osteoarthritis cartilage degeneration by releasing histone deacetylase 4 inhibition of runt-related transcription factor-2 and increasing osteoarthritis-related genes: a novel mechanism of human osteoarthritis cartilage degeneration.

Cao K, Wei L, Zhang Z, Guo L, Zhang C, Li Y, Sun C, Sun X, Wang S, Li P, Wei X - Arthritis Res. Ther. (2014)

HDAC4 downregulates OA-related gene expressions in human OA chondrocytes. Human OA chondrocytes were transfected with HDAC4-flag or HDAC4 siRNA. All transfections were performed in triplicate. Data are presented as means ± SD. *P <0.05. (A) The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 or control vector transfection. (B)The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 siRNA or scrambled siRNA control transfection. HDAC4, histone deacetylase 4; MMP, matrix metalloprotease; OA, osteoarthritis; Runx2, runt-related transcription factor-2; SD, standard deviation; siRNA, small interfering RNA; TSA, trichostatin A.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4265470&req=5

Fig5: HDAC4 downregulates OA-related gene expressions in human OA chondrocytes. Human OA chondrocytes were transfected with HDAC4-flag or HDAC4 siRNA. All transfections were performed in triplicate. Data are presented as means ± SD. *P <0.05. (A) The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 or control vector transfection. (B)The mRNA of HDAC4, Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, type II collagen, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 was tested by real-time PCR after HDAC4 siRNA or scrambled siRNA control transfection. HDAC4, histone deacetylase 4; MMP, matrix metalloprotease; OA, osteoarthritis; Runx2, runt-related transcription factor-2; SD, standard deviation; siRNA, small interfering RNA; TSA, trichostatin A.
Mentions: Our data showed that overexpression of HDAC4 decreased the mRNA levels of Ihh, type X collagen, MMP1, MMP3, MMP-13, IL-1 beta, Cox2, iNos, ADAMTS-4 and -5 in OA chondrocytes to 11%, 23%, 33.8%, 23.7%, 33%, 60%, 57.7%, 54.7%, 45.1% and 48.9%, respectively, of those in controls (Figure 5A). In addition, the mRNA levels of type II collagen and aggrecan significantly increased 4.7- and 9.0-fold after HDAC4 transfection (Figure 5A) (P <0.05). In contrast, knockdown HDAC4 by HDAC4 siRNA showed an opposite effect (Figure 5B) (P <0.05). The transfection efficiencies of HDAC4 and HDAC4 siRNA were validated by real-time PCR (Figure 5A, B).Figure 5

Bottom Line: The levels of HDAC4 in the cartilage from OA patients and healthy 40- to 60-year-old donors were decreased to 31% and 65% compared with specimens from 20- to 40-year-old healthy donors, respectively (P <0.05).In addition, the data also shows that overexpression of HDAC4 not only decreased the expression of interleukin (IL)-1β, Cox2 and iNos and increased the expression of aggrecan, but also partially blocked the effect of IL-1β on expression of catabolic events in human OA chondrocytes.In contrast, inhibition of HDAC4 by TSA drug had an opposite effect.

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: To investigate if decreased histone deacetylase 4 (HDAC4) is associated with human osteoarthritis (OA) cartilage degeneration by releasing HDAC4 inhibition of runt-related transcription factor-2 (Runx2) resulting in increase of OA cartilage degeneration-related genes.

Methods: The mRNA and protein levels of HDAC4, Runx2, matrix metalloproteinase (MMP)-13, Indian hedgehog (Ihh) and type X collagen were detected by performing real-time PCR (RT-PCR), western blotting and immunohistochemistry on specimens from human OA and normal cartilage. To further explore the mechanism of regulation of Runx2 and OA-related genes by HDAC4, changes in these OA-related genes were further quantified by RT-PCR after overexpression of HDAC4 and knockdown of HDAC4 by siRNA. Runx2 and MMP-13 promoter activities were measured by dual luciferase assays.

Results: The levels of HDAC4 in the cartilage from OA patients and healthy 40- to 60-year-old donors were decreased to 31% and 65% compared with specimens from 20- to 40-year-old healthy donors, respectively (P <0.05). Decreased HDAC4 was associated with increased Runx2 and other OA-related genes in human OA cartilage, specifically: MMP-13, Ihh and type X collagen. Exogenous HDAC4 decreased the mRNA levels of Runx2, MMP1, MMP3, MMP-13, type X collagen, Ihh, ADAMTS-4 and -5, and increased the mRNA of type II collagen. In addition, the data also shows that overexpression of HDAC4 not only decreased the expression of interleukin (IL)-1β, Cox2 and iNos and increased the expression of aggrecan, but also partially blocked the effect of IL-1β on expression of catabolic events in human OA chondrocytes. HDAC4 also inhibited Runx2 promoter activity and MMP13 promotor activity in a dose-dependent manner. In contrast, inhibition of HDAC4 by TSA drug had an opposite effect.

Conclusions: Our study is the first to demonstrate that decreased HDAC4 contributes, at least in part, to the pathogenesis of OA cartilage degeneration. Thus, HDAC4 may have chondroprotective properties by inhibiting Runx2 and OA-related genes.

Show MeSH
Related in: MedlinePlus