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The Effect of Repeated Umbilical Cord Occlusions on Pulmonary Surfactant Protein mRNA Levels in the Ovine Fetus

Lin Zhao, MD

Lucy Green, PhD

Fred Possmayer, PhD

Bryan S. Richardson, MD

CIHR Group in Fetal and Neonatal Health and Development, Departments of Obstetrics and Gynacology and of Physiology, The Lawson Health Research Institute, University of Western Ontario, London, Ontario, Canada; Centre for FOAD, Princess Anne Hospital, Southampton, United Kingdom; Departments of Obstetrics and Gynaecology and Physiology, University of Toronto, Toronto, Ontario, Canada

Alan D. Bocking, MD

CIHR Group in Fetal and Neonatal Health and Development, Departments of Obstetrics and Gynacology and of Physiology, The Lawson Health Research Institute, University of Western Ontario, London, Ontario, Canada; Centre for FOAD, Princess Anne Hospital, Southampton, United Kingdom; Departments of Obstetrics and Gynaecology and Physiology, University of Toronto, Toronto, Ontario, Canada abocking{at}mtsinai.on.ca

Objectives: In this study we sought to determine the effect of brief repeated umbilical cord occlusions (rUCO) on surfactant protein (SP) mRNA levels in the fetal sheep lung at two different gestational ages.

Methods: Fourteen fetuses at 112 to 115 days' gestation (control n = 7, rUCO n = 7) and 15 fetuses at 130 to 133 days' gestation (control n = 7, rUCO n = 8) were studied over 4 successive days with rUCO of 90 seconds duration performed every 30 minutes for 3 to 5 hours each day in the rUCO animals. Blood samples were collected for corticotrophin (ACTH) and cortisol measurements. Animals were killed within 1 hour of the final cord occlusion. SP-A, -B, -C, and -D mRNA levels were determined in lung tissue using a ribonuclease protection assay.

Results: Cord occlusions resulted in temporary increases in circulating ACTH on day 1 with both gestational ages, but the elevations were blunted by day 4. Plasma cortisol levels increased transiently with the larger effect being observed on day 4, in particular with the near-term group. With advancing gestational age there was a significant (P <.05) increase in the level of SP-A (control 112-115 days: 0.01 ± 0.01 vs control 130-133 days: 0.07 ± 0.02 fmol/mg RNA), SP -B (control 112-115 days: 0.02 ± 0.01 vs control 130-133 days: 0.07 ± 0.01 fmol/mg RNA) and SP-C (control 112-115 days: 0.13 ± 0.09 vs control 130-133 days: 0.51 ± 0.10 fmol/mg RNA), but not SP-D mRNA levels (control 112-115 days: 0.002 ± 0.002 vs control 130-133 days: 0.01 ± 0.002 fmol/mg RNA). At 112 to 115 days, there was no significant change in any of the SP mRNA levels following rUCO compared to controls. However, the same regime of rUCO at 130 to 133 days resulted in an 85% reduction in SP-A and SP-B mRNA content and a 66% reduction in SP-C mRNA levels compared to controls.

Conclusion: The surprising decrease in SP-A and SP-B mRNA levels, which contrasts with other studies, suggests intermittent asphyxial episodes impact differently on surfactant apoprotein mRNA expression than does prolonged hypoxia.

Key Words: Surfactant protein mRNA • fetus • asphyxia • umbilical cord conclusion

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