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      Vitamin C and sodium bicarbonate enhance the antioxidant ability of H9C2 cells and induce HSPs to relieve heat stress

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          Abstract

          <p class="first" id="Par1">Heat stress is exacerbated by global warming and affects human and animal health, leading to heart damage caused by imbalances in reactive oxygen species (ROS) and the antioxidant system, acid-base chemistry, electrolytes and respiratory alkalosis. Vitamin C scavenges excess ROS, and sodium bicarbonate maintains acid-base and electrolyte balance, and alleviates respiratory alkalosis. Herein, we explored the ability of vitamin C alone and in combination with equimolar sodium bicarbonate (Vitamin C-Na) to stimulate endogenous antioxidants and heat shock proteins (HSPs) to relieve heat stress in H9C2 cells. Control, vitamin C (20 μg/ml vitamin C for 16 h) and vitamin C-Na (20 μg/ml vitamin C-Na for 16 h) groups were heat-stressed for 1, 3 or 5 h. Granular and vacuolar degeneration, karyopyknosis and damage to nuclei and mitochondria were clearly reduced in treatment groups, as were apoptosis, lactate dehydrogenase activity and ROS and malondialdehyde levels, while superoxide dismutase activity was increased. Additionally, CRYAB, Hsp27, Hsp60 and Hsp70 mRNA levels were upregulated at 3 h ( <i>p</i> &lt; 0.01), and protein levels were increased for CRYAB at 0 h ( <i>p</i> &lt; 0.05) and 1 h ( <i>p</i> &lt; 0.01), and for Hsp70 at 3 and 5 h ( <i>p</i> &lt; 0.01). Thus, pre-treatment with vitamin C or vitamin C-Na might protect H9C2 cells against heat damage by enhancing the antioxidant ability and upregulating CRYAB and Hsp70. </p>

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          Most cited references55

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          The ATPase cycle of Hsp90 drives a molecular 'clamp' via transient dimerization of the N-terminal domains.

          How the ATPase activity of Heat shock protein 90 (Hsp90) is coupled to client protein activation remains obscure. Using truncation and missense mutants of Hsp90, we analysed the structural implications of its ATPase cycle. C-terminal truncation mutants lacking inherent dimerization displayed reduced ATPase activity, but dimerized in the presence of 5'-adenylamido-diphosphate (AMP-PNP), and AMP-PNP- promoted association of N-termini in intact Hsp90 dimers was demonstrated. Recruitment of p23/Sba1 to C-terminal truncation mutants also required AMP-PNP-dependent dimerization. The temperature- sensitive (ts) mutant T101I had normal ATP affinity but reduced ATPase activity and AMP-PNP-dependent N-terminal association, whereas the ts mutant T22I displayed enhanced ATPase activity and AMP-PNP-dependent N-terminal dimerization, indicating a close correlation between these properties. The locations of these residues suggest that the conformation of the 'lid' segment (residues 100-121) couples ATP binding to N-terminal association. Consistent with this, a mutation designed to favour 'lid' closure (A107N) substantially enhanced ATPase activity and N-terminal dimerization. These data show that Hsp90 has a molecular 'clamp' mechanism, similar to DNA gyrase and MutL, whose opening and closing by transient N-terminal dimerization are directly coupled to the ATPase cycle.
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            Heat shock proteins: endogenous modulators of apoptotic cell death.

            The highly conserved heat shock proteins (HSPs) accumulate in cells exposed to heat and a variety of other stressful stimuli. HSPs, which function mainly as molecular chaperones, allow cells to adapt to gradual changes in their environment and to survive in otherwise lethal conditions. The events of cell stress and cell death are linked and HSPs induced in response to stress appear to function at key regulatory points in the control of apoptosis. HSPs include antiapoptotic and proapoptotic proteins that interact with a variety of cellular proteins. Their expression level can determine the fate of the cell in response to a death stimulus, and apoptosis-inhibitory HSPs, in particular HSP27 and HSP70, may participate in carcinogenesis. This review summarizes apoptosis-regulatory function of HSPs.
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              Invited review: genes involved in the bovine heat stress response.

              The cellular heat stress (HS) response is one component of the acute systemic response to HS. Gene networks within and across cells and tissues respond to environmental heat loads above the thermoneutral zone with both intra- and extracellular signals that coordinate cellular and whole-animal metabolism. Activation of these systems appears to be initiated at skin surface temperatures exceeding 35 degrees C as animals begin to store heat and rapidly increase evaporative heat loss (EVHL) mechanisms. Gene expression changes include 1) activation of heat shock transcription factor 1 (HSF1); 2) increased expression of heat shock proteins (HSP) and decreased expression and synthesis of other proteins; 3) increased glucose and amino acid oxidation and reduced fatty acid metabolism; 4) endocrine system activation of the stress response; and 5) immune system activation via extracellular secretion of HSP. If the stress persists, these gene expression changes lead to an altered physiological state referred to as "acclimation," a process largely controlled by the endocrine system. In the acclimated state, metabolism is adjusted to minimize detrimental effects of increased thermal heat load. The role of secreted HSP in feedback regulation of the immune and endocrine system has not yet been investigated. The variation in EVHL among animals and the central role that HSF1 has in coordinating thermal tolerance suggest that there is opportunity to improve thermal tolerance via gene manipulation. Determining the basis for altered energy metabolism during thermal stress will lead to opportunities for improved animal performance via altered nutritional management.
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                Author and article information

                Journal
                Cell Stress and Chaperones
                Cell Stress and Chaperones
                Springer Nature America, Inc
                1355-8145
                1466-1268
                July 2018
                February 13 2018
                July 2018
                : 23
                : 4
                : 735-748
                Article
                10.1007/s12192-018-0885-2
                6045543
                29442224
                f362775a-a66e-46e1-af83-a810eecb40f0
                © 2018

                http://www.springer.com/tdm

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