<p><b>OBJECTIVE</b>To investigate the effect of recombinant
adenovirus-mediated
heat shock protein 70 (HSP70) on
energy metabolism of
mitochondria in intestinal
epithelial cells (IEC-6) after
hypoxia/reoxygenation
injury .</p><p><b>
METHODS</b>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 blotting. 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
hypoxia/reoxygenation after Ad-HSP70
transfection). The activity of mitochondrial
dehydrogenase was assessed by MTf
method. The contents of cellular
ATP,
ADP , AMP and energy
charge (EC)were determined by
high-performance liquid chromatography (
HPLC).</p><p><b>RESULTS</b>The expression of HSP70
protein in IEC-6
cells was significantly upregulated after Ad-HSP70
transfection compared with empty
adenovirus vector
transfection. Compared with that in
control group, the activity of mitochondrial
dehydrogenase was significantly lowered in IEC-6
cells in
hypoxia/reoxygenation group (P < 0.01). The activity of mitochondrial
dehydrogenase in Ad-HSP70
transfection group was significantly 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
hypoxia/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 < 0.050 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).</p><p><b>CONCLUSION</b>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.
Mitochondria may be one of main target
organelles for HSP70 in
protection of IEC against
hypoxia/reoxygenation
injury.</p>