Manganese superoxide dismutase expression in alveolar type II epithelial cells from nonventilated and hypoperfused lungs.
Lungs that have been hypoxic and hypoperfused because of atelectasis and the resulting decrease in pulmonary arterial blood flow develop specific decreases in manganese superoxide dismutase (MnSOD) activity and are sensitive to oxidant injury during reoxygenation. Since the MnSOD protein is concentrated in mitochondria of alveolar epithelial type II cells (ATII), we hypothesized that expression of MnSOD would be decreased in these cells also as a result of hypoxia. To investigate whether regulation of MnSOD expression occurred before or after transcription, we determined whether MnSOD protein content or steady-state mRNA level changed after hypoxia as well. ATII cells were isolated by elastase digestion from lungs of adult rabbits after right lungs had been hypoxic and hypoperfused for 7 days because of unilateral atelectasis. MnSOD activity was measured by inhibition of cytochrome c reduction in the presence of 1 mM KCN, MnSOD protein content was measured on immunoblots, and MnSOD mRNA was quantified on slot blot autoradiograms. MnSOD activity was 8.4 +/- 1.9 U/mg protein in ATII cells from control lungs and 6.8 +/- 1.5 U/mg protein in ATII cells from hypoxic and hypoperfused lungs (n = 9, P = 0.037). MnSOD protein content was 5.1 +/- 1.4 micrograms/mg protein in ATII cells from control and 4.1 +/- 1.2 micrograms/mg protein in ATII cells from hypoxic and hypoperfused lungs (P = 0.021). ATII cell MnSOD mRNA/18S ribosomal RNA (ratio of arbitrary absorbance units) determined by RNA slot blots was 2.18 +/- 1.26 in ATII cells from control lungs and 2.94 +/- 0.88 in ATII cells from hypoxic lungs (n = 7, P > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Russell
W J
WJ
Birmingham VA Medical Center, Alabama.
Matalon
S
S
Jackson
R M
RM
D000818:Animals; D004847:Epithelial Cells; D015870:Gene Expression; D000860:Hypoxia / Q000201:enzymology*; D008297:Male; D011650:Pulmonary Alveoli / Q000166:cytology / Q000201:enzymology*; D012333:RNA, Messenger / Q000032:analysis; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D011817:Rabbits; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D013482:Superoxide Dismutase