TNF-alpha decreases hsp 27 in human blood mononuclear cells: involvement of protein kinase c.
ABSTRACT Treatment of PBMCs with TNF-alpha decreased the levels of heat shock protein (HSP) 27, but had little effect on the level of HSP70. Parallel to the decrease of HSP27, TNF-alpha increased the level of HSP27 in the incubation medium of the cells. The decrease of HSP27 induced by TNF-alpha was suppressed by the pretreatment of PBMCs with the specific protein kinase C (PKC) inhibitor, GF109203X. Furthermore, phorbol myristate acetate (PMA), a PKC stimulant, but not dibutyryl cyclic AMP, a protein kinase A stimulant, decreased the levels of HSP27. To investigate the effect of TNF-alpha on the oligomerization state of HSP27 in PBMCs, we performed sucrose density gradient centrifugation with subsequent fractionation and immunoassay. Extract of vehicle-treated PBMCs contained mainly dissociated forms of HSP27. The amounts of dissociated forms of HSP27 in PBMCs was decreased by TNF-alpha, while the amounts of aggregated form of HSP27 was little changed. In intact PBMCs, HSP27 is constitutively phosphorylated at Ser78, but not at Ser15 or at Ser82. The amount of phosphorylated HSP27 at Ser78 was decreased by TNF-alpha. These results indicate that TNF-alpha reduces HSP27 in PBMCs through PKC activation. This decrease may be due to efflux of dissociated form of HSP27, phosphorylated HSP27 at Ser78, from the cells.
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ABSTRACT: Cholesteatoma is a destructive and expanding growth of keratinizing squamous epithelium in the middle ear or petrous apex. The molecular and cellular processes of the pathogenesis of acquired middle ear cholesteatoma have not been fully understood. In this study, comparative proteomic analysis was conducted to investigate the roles of specific proteins in the pathways regarding keratinocyte proliferation in cholesteatoma. The differential proteins were detected by comparing the two-dimension electrophoresis (2-DE) maps of the epithelial tissues of 12 attic cholesteatomas with those of retroauricular skins. There were 14 upregulated proteins in the epithelial tissues of cholesteatoma in comparison with retroauricular skin. The modulation of five crucial proteins, HSP27, PRDX2, GRP75, GRP78 and GRP94, was further determined by RT-PCR, Western blot and immunohistochemistry. Phosphorylation of HSP27 at Ser-82 was identified by mass spectroscopy. The results of this study suggested that phosphorylated HSP27 is the end expression of two potential signal-transduction pathways, and together with PRDX2, they are very likely involved in the proliferation of keratinocytes in cholesteatoma. Upregulations of GRP75, GRP78 and GRP94 in keratinocytes may be able to counter endoplasmic reticulum stress, to inhibit cell apoptosis, to prevent protein unfolding and to promote cholesteatoma growth.International Journal of Molecular Sciences 07/2013; 14(7):14439-14459. · 2.34 Impact Factor
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ABSTRACT: Heat shock protein (HSP) 27 is related to the pathogenesis of AF. However, the clinical relationship between HSP27 and AF is unclear. The present study was conducted to determine the clinical relationship between HSP27 and atrial fibrillation (AF). A case-control study was conducted (AF, n=114; control, n=100). Serum HSP27 (HSP27S) levels were measured by ELISA, and its correlations with electrophysiological characteristics and catheter ablation outcomes were investigated. The patients with AF had a larger left atrial diameter (LAD), waist circumference, and body mass index, and a lower baseline HSP27S level, than controls. After logistic multivariate analysis, low baseline HSP27S was independently associated with AF. In patients with AF, those with paroxysmal AF (PAF) had higher baseline HSP27S levels compared with those without PAF. In patients with PAF, lower baseline HSP27S was associated with larger LAD, whereas baseline HSP27S was not correlated with LAD in controls. In PAF, low baseline HSP27S (≤3.85 ng/mL) was associated with low atrial voltage and nonpulmonary vein ectopies. In non-PAF, the mean fractionated interval had a good correlation with baseline HSP27S. After catheter ablation, a high baseline HSP27S level could predict sinus rhythm maintenance in the patients with PAF. Baseline HSP27S was also correlated with interleukin 10 and tumor necrosis factor-α levels. Analysis of buffy coat mRNA levels showed the same correlations. The HSP27S levels were correlated with LAD, left atrial voltage, and fractionated intervals, and predicted AF recurrence after catheter ablation. The mechanisms could be related to inflammation.Circulation Arrhythmia and Electrophysiology 02/2012; 5(2):334-40. · 5.95 Impact Factor
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ABSTRACT: Heat shock proteins (HSPs) are molecular chaperones that facilitate the proper folding and assembly of nascent polypeptides and assist in the refolding and stabilization of damaged polypeptides. Through these largely intracellular functions, the HSPs maintain homeostasis and assure cell survival. However, a growing body of literature suggests that HSPs have important effects in the extracellular environment as well. Extracellular HSPs are released from damaged or stressed cells and appear to act as local "danger signals" that activate stress response programs in surrounding cells. Importantly, extracellular HSPs have been shown to activate the host innate and adaptive immune response. With this in mind, extracellular HSPs are commonly included in a growing list of a family of proteins known as danger-associated molecular patterns (DAMPs) or alarmins, which trigger an immune response to tissue injury, such as may occur with trauma, ischemia-reperfusion injury, oxidative stress, etc. Extracellular HSPs, including Hsp72 (HSPA), Hsp27 (HSPB1), Hsp90 (HSPC), Hsp60 (HSPD), and Chaperonin/Hsp10 (HSPE) are especially attractrive candidates for DAMPs or alarmins which may be particularly relevant in the pathophysiology of the sepsis syndrome.The Open Inflammation Journal 10/2011; 4:49-60.