OBJECTIVETo assess the expression of Hsp72 and Hsp73 in chronic obstructive pulmonary disease (COPD) and to evaluate their roles in damage from coal dust exposure.

METHODSA case control study of 50 coal miners suffering from COPD and 50 healthy coal miners were selected from one coal mine. The levels of Hsp72 and Hsp73 in peripheral blood lymphocytes were determined by flow cytometry for all subjects.

RESULTS(1) The expression of basic Hsp72 of peripheral blood lymphocytes for patients and controls was not different from that inducible expressed Hsp72 by 42 degrees C heat stress or by BPDE exposure. (2) The expression of Hsp72 in COPD patients (17.7 +/- 4.9) was significantly lower than that in healthy coal miners (22.6 +/- 10.0) (P < 0.01). On the other hand, the expression of Hsp73 in COPD patients (33.5 +/- 11.7) was higher than that in healthy coal miners (19.6 +/- 5.9) (P < 0.01). (3) A-positive relationship between the expression of Hsp72 and cumulative inhaling coal dust exposure was observed. No relationship was found between Hsp73 and cumulative inhaling coal dust exposure.

CONCLUSIONThe decreased expressions of Hsp72 in peripheral blood lymphocytes of COPD coal miners.

Aged ; Case-Control Studies ; Coal Mining ; Dust ; Flow Cytometry ; HSC70 Heat-Shock Proteins ; biosynthesis ; HSP72 Heat-Shock Proteins ; biosynthesis ; Humans ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Occupational Exposure ; Pulmonary Disease, Chronic Obstructive ; metabolism

AIMTo investigate the mechanism of protection by sound conditioning from acoustic trauma.

METHODSSound conditioning experimental model of animals was established. The expression of CaM, HSP70 and F-actin in hair cells were examined with the method of immunohistochemistry. Free calcium concentration in hair cells was observed by LSCM at the same time. Quantitative investigation was devised to assess the changes of F-actin, CaM, HSP70 and intracellular calcium concentration in hair cells.

RESULTSThe expression of CaM, HSP70 and F-actin all showed an increased trend after noise exposure. HSP70 and F-actin expressed significantly more in group CH than that expressed in group H. Compared with group H, the expression of CaM showed an increased trend in group CH. Elevation of intracellular calcium concentration could be resulted from noise exposure. The calcium concentration in group H was significantly higher than that in group C and group CH.

CONCLUSIONA suitable sound conditioning can make the auditory system of guinea pig more resistant to noise trauma. The strengthened cytoskeleton system and the intracellular calcium homeostasis play a critical role in the protective mechanism of sound conditioning.

Acclimatization ; Actins ; metabolism ; Animals ; Auditory Threshold ; Calcium ; metabolism ; Calmodulin ; metabolism ; Cytoskeleton ; Disease Models, Animal ; Female ; Guinea Pigs ; HSC70 Heat-Shock Proteins ; metabolism ; Hair Cells, Auditory ; cytology ; metabolism ; Hearing Loss, Noise-Induced ; pathology ; Male

OBJECTIVETo investigate the anti-atherotic effects of heme-L-lysinate in a rabbit model of atherosclerosis and related machanisms.

METHODSAdult rabbits were treated with 1% cholesterol diet (chol group, n = 8) or 1% cholesterol diet plus heme-L-lysinate (9 mgxkg(-1)xd(-1), Heme group, n = 8) or 1% cholesterol diet plus isotonic Na chloride (Na chloride group, n = 8) for 10 weeks. Eight rabbits fed with normal diet served as normal control. Aortic carbon monoxide (CO) was quantified spectrophotometrically by the formation of carboxyhaemoglobin (HbCO). Aortic heme oxygenase-1 (HO-1) and HSP70 mRNA and protein expressions were detected by RT-PCR and immunohistochemical staining.

RESULTSAortic CO production and HO-1 activity were significantly increased in chol group and Na chloride group compared those in normal control group (P < 0.01). Aortic plaque area was significantly reduced in heme group (26.6% +/- 9.2%) than that in chol group (42.3% +/- 8.7%, P < 0.01). Aortic HO-1 expression, CO production and HSP70 were significantly increased in heme group than those in chol group and Na chloride group (all P < 0.01).

CONCLUSIONSHeme-L-lysinate could attenuate atherosclerotic progression through upregulating HO-1 and HSP70 expression and increasing CO production in this model.

Animals ; Atherosclerosis ; metabolism ; prevention & control ; Carbon Monoxide ; metabolism ; Cholesterol, Dietary ; Diet, Atherogenic ; Disease Models, Animal ; HSC70 Heat-Shock Proteins ; genetics ; metabolism ; Heme ; analogs & derivatives ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Lysine ; analogs & derivatives ; pharmacology ; Male ; RNA, Messenger ; genetics ; Rabbits

In response to viral infection, RIG-I-like RNA helicases detect viral RNA and signal through the mitochondrial adapter protein VISA. VISA activation leads to rapid activation of transcription factors IRF3 and NF-κB, which collaborate to induce transcription of type I interferon (IFN) genes and cellular antiviral response. It has been demonstrated that VISA is activated by forming prion-like aggregates. However, how this process is regulated remains unknown. Here we show that overexpression of HSC71 resulted in potent inhibition of virus-triggered transcription of IFNB1 gene and cellular antiviral response. Consistently, knockdown of HSC71 had opposite effects. HSC71 interacted with VISA, and negatively regulated virus-triggered VISA aggregation. These findings suggest that HSC71 functions as a check against VISA-mediated antiviral response.
Adaptor Proteins, Signal Transducing ; biosynthesis ; chemistry ; genetics ; metabolism ; Cell Aggregation ; genetics ; GPI-Linked Proteins ; metabolism ; Gene Knockdown Techniques ; HEK293 Cells ; HSC70 Heat-Shock Proteins ; genetics ; metabolism ; Heat-Shock Response ; genetics ; Humans ; Interferon Regulatory Factor-3 ; genetics ; metabolism ; Interferon-beta ; genetics ; NF-kappa B ; genetics ; Prions ; metabolism ; Receptors, Retinoic Acid ; metabolism ; Viruses ; drug effects ; metabolism ; pathogenicity

Histone deacetylase 6 (HDAC6), a predominantly cytoplasmic protein deacetylase, participates in a wide range of cellular processes through its deacetylase activity. However, the diverse functions of HDAC6 cannot be fully elucidated with its known substrates. In an attempt to explore the substrate diversity of HDAC6, we performed quantitative proteomic analyses to monitor changes in the abundance of protein lysine acetylation in response to HDAC6 deficiency. We identified 107 proteins with elevated acetylation in the liver of HDAC6 knockout mice. Three cytoplasmic proteins, including myosin heavy chain 9 (MYH9), heat shock cognate protein 70 (Hsc70), and dnaJ homolog subfamily A member 1 (DNAJA1), were verified to interact with HDAC6. The acetylation levels of these proteins were negatively regulated by HDAC6 both in the mouse liver and in cultured cells. Functional studies reveal that HDAC6-mediated deacetylation modulates the actin-binding ability of MYH9 and the interaction between Hsc70 and DNAJA1. These findings consolidate the notion that HDAC6 serves as a critical regulator of protein acetylation with the capability of coordinating various cellular functions.
Acetylation ; Actins ; chemistry ; metabolism ; Animals ; Cell Line ; Chromatography, High Pressure Liquid ; HSC70 Heat-Shock Proteins ; metabolism ; HSP40 Heat-Shock Proteins ; metabolism ; Histone Deacetylase 6 ; Histone Deacetylases ; metabolism ; Isotope Labeling ; Liver ; metabolism ; Lysine ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Microscopy, Confocal ; Nonmuscle Myosin Type IIA ; metabolism ; Protein Binding ; Proteomics ; Substrate Specificity ; Tandem Mass Spectrometry

PURPOSE: The mechanisms underlying repolarization abnormalities during pregnancy are not fully understood. Although maternal serotonin (5-hydroxytryptamine, 5-HT) production is an important determinant for normal fetal development in mice, its role in mothers remains unclear. We evaluated the role of serotonin in ventricular repolarization in mice hearts via 5Htr3 receptor (Htr3a) and investigated the mechanism of QT-prolongation during pregnancy. MATERIALS AND METHODS: We measured current amplitudes and the expression levels of voltage-gated K⁺ (Kv) channels in freshly-isolated left ventricular myocytes from wild-type non-pregnant (WT-NP), late-pregnant (WT-LP), and non-pregnant Htr3a homozygous knockout mice (Htr3a(−/−)-NP). RESULTS: During pregnancy, serotonin and tryptophan hydroxylase 1, a rate-limiting enzyme for the synthesis of serotonin, were markedly increased in hearts and serum. Serotonin increased Kv current densities concomitant with the shortening of the QT interval in WT-NP mice, but not in WT-LP and Htr3a(−/−)-NP mice. Ondansetron, an Htr3 antagonist, decreased Kv currents in WT-LP mice, but not in WT-NP mice. Kv4.3 directly interacted with Htr3a, and this binding was facilitated by serotonin. Serotonin increased the trafficking of Kv4.3 channels to the cellular membrane in WT-NP. CONCLUSION: Serotonin increases repolarizing currents by augmenting Kv currents. Elevated serotonin levels during pregnancy counterbalance pregnancy-related QT prolongation by facilitating Htr3-mediated Kv currents.
*Action Potentials/drug effects ; Animals ; Cell Membrane/drug effects/metabolism ; Disease Models, Animal ; Electrocardiography ; Female ; HSC70 Heat-Shock Proteins/metabolism ; HSP90 Heat-Shock Proteins/metabolism ; Heart Ventricles/drug effects/*metabolism ; Mice, Inbred C57BL ; Mice, Knockout ; Myocytes, Cardiac/drug effects/metabolism ; Potassium Channels/metabolism ; Pregnancy ; Rabbits ; Rats, Sprague-Dawley ; Receptors, Serotonin, 5-HT3/metabolism ; Serotonin/*metabolism ; Serotonin 5-HT3 Receptor Agonists/pharmacology

OBJECTIVETo study the protective effect of puerarin on MPP(+) -induced SH-SY5Y cells by chaperone-mediated autophagy (CMA).

METHODThe Parkinson's disease cell model was established by injuring SH-SY5Y cells with 1 mmol x L(-1) MPP+. The CCK-8 staining was adopted to detect the effect the puerarin of different concentrations on the survival rate of MPP(+)-induced SH-SYSY cells. The autophagosome formation was observed under transmission electron microscope. The AO staining showed the changes in the lysosome activity. RT-PCR was used to detect the changes in Lamp2a and Hsc70 mRNA expressions. The western blotting was adopted to test the expressions of Lamp2a, Hsc70 and alpha-synuclein protein in cells.

RESULTWithin the concentration range of 12. 5-50.0 micromol x L(-1), the pretreatment with puerain for 30 minutes could protect the injury of MPP+ in SH-SY5Y cells, and showed a certain dose-effect relationship. The AO staining and electron microscope showed the effect of puerain within the concentration range of 12.5-50.0 micromol x L(-1) on 1 mmol x L(-1) MPP(+)-induced SH-SY5Y cells; autophagosomes emerged in cells, and increased along with the rise in the puerarin dose. The results of the flow cytometry revealed that 50.0 micromol x L(-1) of puerarin could protect against the increase of the ROS level in 1 mmol x L(-1) MPP(+) -induced SH-SY5Y cells and prevent the oxidative injury. The results of RT-PCR and western blotting indicated that puerain within the concentration range of 12.5-50.0 micromol x L(-1) alleviated the MPP(+)-induced SH-SY5Y cell injury, and inhibited the accumulation of alpha-synuclein proteins in MPP(+) -induced SH-SY5Y cells by up-regulating Hsc70, Lamp2a mRNA and protein level.

CONCLUSIONPuerarin could protect against the MPP(+) -induced cell injury, whose protective mechanism may be related to the chaperone-mediated autophagy pathway of interventional molecules.

Autophagy ; drug effects ; genetics ; HSC70 Heat-Shock Proteins ; genetics ; Humans ; Isoflavones ; pharmacology ; Lysosomal-Associated Membrane Protein 2 ; genetics ; Molecular Chaperones ; genetics ; Parkinson Disease ; drug therapy ; genetics ; Phagosomes ; drug effects ; genetics ; Piperidines ; pharmacology ; Pyrazoles ; pharmacology ; Tumor Cells, Cultured ; Up-Regulation ; drug effects ; genetics