Influence of HSP70 on function and energy metabolism of mitochondria in intestinal epithelial cells after hypoxia/reoxygenation
Author(s): YUAN Zhi-qiang, LI Xiao-lu, PENG Yi-zhi, WANG Pei, HUANG Yue-sheng, YANG Zong-cheng
Pages: 203-206
Year: 2008 Issue: 3
Journal: CHINESE JOURNAL OF BURNS
Keyword: Heat shock proteins; Intestines; Epithelial cells; Energy metabolism; Mitochon-dria; Cell hypoxia;
Abstract: Objective To investigate the effect of recombinant adenovirus-mediated heat shock pro-tein 70(HSP70)on energy metabolism of mitochondria in intestinal epithelial cells(IEC-6)after hypoxia/reoxygenation injury. Methods IEC-6 cells were transfected with HSP70 recombinant adenovirus vectors (Ad-HSP70)and empty adenovirus vectors.The expression of HSP70 protein was detected by Western blot-ting.Cultured IEC-6 cells were divided into:control group(without treatment),hypoxia/reoxygenation group(with challenge of hypoxia/reoxygenation)and Ad-HSP70 transfection group(with challenge of hy-poxia/reoxygenation after Ad-HSP70 transfection).The activity of mitochondrial dehydrogenase was assessed by MTT method.The contents of cellular ATP,ADP,AMP and energy charge(EC)were determined by high-performance liquid chromatography(HPLC). Results The expression of HSP70 protein in IEC-6 cells was significantly upregulated after Ad-HSP70 transfection compared with empty adenovirus vector trans-fection.Compared with that in control group,the activity of mitochondrial dehydrogenase was significantly low-ered in IEC-6 cells in hypoxia/reoxygenation group(P＜0.01).The activity of mitochondrial dehydrogenase in Ad-HSP70 transfection group was significantiy greater than that in hypoxia/reoxygenation group ( P＜0.01).Compared with those in control group,the content of cellular ATP was significantly decreased in hy-poxia/reoxygenation group,the contents of cellular ADP and AMP were significantly increased.The above cell energy indices in Ad-HSP70 transfection group was similar to those in control group(P＞0.05).which were ameliorated compared with those in hypoxia/reoxygenation group(P＜0.05 or P＜0.01).The cellular EC in hypoxia/reoxygenation group(0.615±0.060)was significantly lower than that in control group (0.748±0.012.P<0.01)and Ad-HSP70 transfection group(0.736±0.028,P＜0.01). Conclusion Ad-HSP70 transfection in IEC-6 cells can upregulate the expression of HSP70,the content of cellular ATP and EC after hypoxia/reoxygenation.and protect mitochondrial function.Mitoehondfia may be one of main target organelles for HSP70 in protection of IEC against hypoxia/reoxygenation injury.
Pages: 203-206
Year: 2008 Issue: 3
Journal: CHINESE JOURNAL OF BURNS
Keyword: Heat shock proteins; Intestines; Epithelial cells; Energy metabolism; Mitochon-dria; Cell hypoxia;
Abstract: Objective To investigate the effect of recombinant adenovirus-mediated heat shock pro-tein 70(HSP70)on energy metabolism of mitochondria in intestinal epithelial cells(IEC-6)after hypoxia/reoxygenation injury. Methods IEC-6 cells were transfected with HSP70 recombinant adenovirus vectors (Ad-HSP70)and empty adenovirus vectors.The expression of HSP70 protein was detected by Western blot-ting.Cultured IEC-6 cells were divided into:control group(without treatment),hypoxia/reoxygenation group(with challenge of hypoxia/reoxygenation)and Ad-HSP70 transfection group(with challenge of hy-poxia/reoxygenation after Ad-HSP70 transfection).The activity of mitochondrial dehydrogenase was assessed by MTT method.The contents of cellular ATP,ADP,AMP and energy charge(EC)were determined by high-performance liquid chromatography(HPLC). Results The expression of HSP70 protein in IEC-6 cells was significantly upregulated after Ad-HSP70 transfection compared with empty adenovirus vector trans-fection.Compared with that in control group,the activity of mitochondrial dehydrogenase was significantly low-ered in IEC-6 cells in hypoxia/reoxygenation group(P＜0.01).The activity of mitochondrial dehydrogenase in Ad-HSP70 transfection group was significantiy greater than that in hypoxia/reoxygenation group ( P＜0.01).Compared with those in control group,the content of cellular ATP was significantly decreased in hy-poxia/reoxygenation group,the contents of cellular ADP and AMP were significantly increased.The above cell energy indices in Ad-HSP70 transfection group was similar to those in control group(P＞0.05).which were ameliorated compared with those in hypoxia/reoxygenation group(P＜0.05 or P＜0.01).The cellular EC in hypoxia/reoxygenation group(0.615±0.060)was significantly lower than that in control group (0.748±0.012.P<0.01)and Ad-HSP70 transfection group(0.736±0.028,P＜0.01). Conclusion Ad-HSP70 transfection in IEC-6 cells can upregulate the expression of HSP70,the content of cellular ATP and EC after hypoxia/reoxygenation.and protect mitochondrial function.Mitoehondfia may be one of main target organelles for HSP70 in protection of IEC against hypoxia/reoxygenation injury.
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