2011PubMedRequires access

[Ethyl pyruvate inhibited HMGB1 expression induced by LPS in macrophages].

Xiao-xing Tian, Chuanxin Wu, Hang Sun, Jianping Gong, Qi Liu, Hui Guo

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Abstract

AIM: To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)expression and releasing in macrophage induced by lipopolysaccharide(LPS). METHODS: The murine macrophage-like cell line RAW264.7 cultured in vitro divided into LPS group and LPS+EP group. The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR. The cytoplasmic and nuclear HMGB1 levels were detected by Western blot. The contents of HMGB1 and TNF-α and IL-6 protein in cultured cells supernatant were detected by ELISA. Immunocytochemistry and confocal laser-scanning microscopy were used to confirm the relocation and distribution of intracellular HMGB1 protein in RAW264.7 cells. RESULTS: HMGB1 mRNA expression in the LPS+EP group was significantly lower than in LPS alone, at 24, 36 and 48 hours. In the LPS+EP stimulation group, the cytoplasm stained weakly while the nuclear stain was stronger than that of the LPS group at the same time points. Both TNF-α and IL-6 levels in LPS+EP group were significantly lower than those in the LPS group at the same time points. EP also effectively prevented the release of HMGB1 protein. CONCLUSION: EP inhibits HMGB1 expression and release from LPS-stimulated macrophages.

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What this paper is about

AIM: To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)expression and releasing in macrophage induced by lipopolysaccharide(LPS). METHODS: The murine macrophage-like cell line RAW264.7 cultured in vitro divided into LPS group and LPS+EP group. The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR. The cytoplasmic and nuclear HMGB1 levels were detected by Western blot. The contents of HMGB1 and TNF-α and IL-6 protein in cultured cells supernatant were detected by ELISA. Immunocytochemistry and confocal laser-scanning microscopy were used to confirm the relocation and distribution of intracellular HMGB1 protein in RAW264.7 cells. RESULTS: HMGB1 mRNA expression in the LPS+EP group was significantly lower than in LPS alone, at 24, 36 and 48 hours. In the LPS+EP stimulation group, the cytoplasm stained weakly while the nuclear stain was stronger than that of the LPS group at the same time points. Both TNF-α and IL-6 levels in LPS+EP group were significantly lower than those in the LPS group at the same time points. EP also effectively prevented the release of HMGB1 protein. CONCLUSION: EP inhibits HMGB1 expression and release from LPS-stimulated macrophages.

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Available abstract

AIM: To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)expression and releasing in macrophage induced by lipopolysaccharide(LPS). METHODS: The murine macrophage-like cell line RAW264.7 cultured in vitro divided into LPS group and LPS+EP group. The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR. The cytoplasmic and nuclear HMGB1 levels were detected by Western blot. The contents of HMGB1 and TNF-α and IL-6 protein in cultured cells supernatant were detected by ELISA. Immunocytochemistry and confocal laser-scanning microscopy were used to confirm the relocation and distribution of intracellular HMGB1 protein in RAW264.7 cells. RESULTS: HMGB1 mRNA expression in the LPS+EP group was significantly lower than in LPS alone, at 24, 36 and 48 hours. In the LPS+EP stimulation group, the cytoplasm stained weakly while the nuclear stain was stronger than that of the LPS group at the same time points. Both TNF-α and IL-6 levels in LPS+EP group were significantly lower than those in the LPS group at the same time points. EP also effectively prevented the release of HMGB1 protein. CONCLUSION: EP inhibits HMGB1 expression and release from LPS-stimulated macrophages.

Key concepts: HMGB1, Lipopolysaccharide, High-mobility group, Molecular biology, Western blot, Cytoplasm, Tumor necrosis factor alpha, Cell culture

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