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Glycolysis-dependent histone deacetylase 4 degradation regulates inflammatory cytokine production.

Wang B, Liu TY, Lai CH, Rao YH, Choi MC, Chi JT, Dai JW, Rathmell JC, Yao TP - Mol. Biol. Cell (2014)

Bottom Line: Inhibition of GSK3β or iNOS suppresses nitric oxide (NO) production, glycolysis, and HDAC4 degradation.We present evidence that sustained glycolysis induced by LPS treatment activates caspase-3, which cleaves HDAC4 and triggers its degradation.Of importance, a caspase-3-resistant mutant HDAC4 escapes LPS-induced degradation and prolongs inflammatory cytokine production.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cancer Biology, Duke University, Durham, NC 27710 Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.

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HDAC4 is required for inflammatory cytokines production and p70 S6K in activated BV2 cells. (A, B) Control and HDAC4 KD BV2 cells were treated with LPS (1 μg/ml) for indicated times. Medium was collected and analyzed for secreted IL-6 and TNF-α by ELISA. (C, D) Cell lysates were subject to ELISA to determine intracellular IL-6 and TNF-α production. Note that LPS-induced secreted and intracellular IL-6 and TNF-α were both much reduced in HDAC4-KD BV2 cells compared with control cells (**p < 0.01 vs. control). (E, F) BV2 cells treated by LPS were harvested at indicated time points. Total RNAs were extracted, and real-time PCR was performed for IL-6 and TNF-α. (G) BV2 cells were treated with LPS at indicated time points, and cell lysates were subject to immunoblotting with antibodies for HDAC4, phosphorylated Erk1/2, JNK, p38, and p70 S6K (T389) as indicated.
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Figure 1: HDAC4 is required for inflammatory cytokines production and p70 S6K in activated BV2 cells. (A, B) Control and HDAC4 KD BV2 cells were treated with LPS (1 μg/ml) for indicated times. Medium was collected and analyzed for secreted IL-6 and TNF-α by ELISA. (C, D) Cell lysates were subject to ELISA to determine intracellular IL-6 and TNF-α production. Note that LPS-induced secreted and intracellular IL-6 and TNF-α were both much reduced in HDAC4-KD BV2 cells compared with control cells (**p < 0.01 vs. control). (E, F) BV2 cells treated by LPS were harvested at indicated time points. Total RNAs were extracted, and real-time PCR was performed for IL-6 and TNF-α. (G) BV2 cells were treated with LPS at indicated time points, and cell lysates were subject to immunoblotting with antibodies for HDAC4, phosphorylated Erk1/2, JNK, p38, and p70 S6K (T389) as indicated.

Mentions: To determine whether HDAC4 is involved inflammatory cytokine production in macrophages, we knocked down HDAC4 in a microglia cell line (BV2) by a small interfering RNA (siRNA) and/or short hairpin RNA (shRNA; Figure 1G; see Materials and Methods). Microglia are resident macrophages in the CNS and play important roles in neuroinflammation (Perry and Gordon, 1988). As shown in Figure 1, A and B, HDAC4-knockdown (KD) BV2 cells secreted much less interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) than control cells (by ∼30–50%) upon LPS treatment. Intracellular IL-6 and TNF-α protein levels were also significantly decreased in HDAC4 KD cells, indicating that synthesis of cytokines is reduced (Figure 1, C and D). The induction of IL-6 and TNF-α mRNA by LPS, however, was not affected by HDAC4 KD (Figure 1, E and F). These results indicate that HDAC4 regulates inflammatory cytokine production posttranscriptionally, possibly at the translation level.


Glycolysis-dependent histone deacetylase 4 degradation regulates inflammatory cytokine production.

Wang B, Liu TY, Lai CH, Rao YH, Choi MC, Chi JT, Dai JW, Rathmell JC, Yao TP - Mol. Biol. Cell (2014)

HDAC4 is required for inflammatory cytokines production and p70 S6K in activated BV2 cells. (A, B) Control and HDAC4 KD BV2 cells were treated with LPS (1 μg/ml) for indicated times. Medium was collected and analyzed for secreted IL-6 and TNF-α by ELISA. (C, D) Cell lysates were subject to ELISA to determine intracellular IL-6 and TNF-α production. Note that LPS-induced secreted and intracellular IL-6 and TNF-α were both much reduced in HDAC4-KD BV2 cells compared with control cells (**p < 0.01 vs. control). (E, F) BV2 cells treated by LPS were harvested at indicated time points. Total RNAs were extracted, and real-time PCR was performed for IL-6 and TNF-α. (G) BV2 cells were treated with LPS at indicated time points, and cell lysates were subject to immunoblotting with antibodies for HDAC4, phosphorylated Erk1/2, JNK, p38, and p70 S6K (T389) as indicated.
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Related In: Results  -  Collection

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Figure 1: HDAC4 is required for inflammatory cytokines production and p70 S6K in activated BV2 cells. (A, B) Control and HDAC4 KD BV2 cells were treated with LPS (1 μg/ml) for indicated times. Medium was collected and analyzed for secreted IL-6 and TNF-α by ELISA. (C, D) Cell lysates were subject to ELISA to determine intracellular IL-6 and TNF-α production. Note that LPS-induced secreted and intracellular IL-6 and TNF-α were both much reduced in HDAC4-KD BV2 cells compared with control cells (**p < 0.01 vs. control). (E, F) BV2 cells treated by LPS were harvested at indicated time points. Total RNAs were extracted, and real-time PCR was performed for IL-6 and TNF-α. (G) BV2 cells were treated with LPS at indicated time points, and cell lysates were subject to immunoblotting with antibodies for HDAC4, phosphorylated Erk1/2, JNK, p38, and p70 S6K (T389) as indicated.
Mentions: To determine whether HDAC4 is involved inflammatory cytokine production in macrophages, we knocked down HDAC4 in a microglia cell line (BV2) by a small interfering RNA (siRNA) and/or short hairpin RNA (shRNA; Figure 1G; see Materials and Methods). Microglia are resident macrophages in the CNS and play important roles in neuroinflammation (Perry and Gordon, 1988). As shown in Figure 1, A and B, HDAC4-knockdown (KD) BV2 cells secreted much less interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) than control cells (by ∼30–50%) upon LPS treatment. Intracellular IL-6 and TNF-α protein levels were also significantly decreased in HDAC4 KD cells, indicating that synthesis of cytokines is reduced (Figure 1, C and D). The induction of IL-6 and TNF-α mRNA by LPS, however, was not affected by HDAC4 KD (Figure 1, E and F). These results indicate that HDAC4 regulates inflammatory cytokine production posttranscriptionally, possibly at the translation level.

Bottom Line: Inhibition of GSK3β or iNOS suppresses nitric oxide (NO) production, glycolysis, and HDAC4 degradation.We present evidence that sustained glycolysis induced by LPS treatment activates caspase-3, which cleaves HDAC4 and triggers its degradation.Of importance, a caspase-3-resistant mutant HDAC4 escapes LPS-induced degradation and prolongs inflammatory cytokine production.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Cancer Biology, Duke University, Durham, NC 27710 Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.

Show MeSH
Related in: MedlinePlus