Insulin inhibits surfactant protein A and B gene expression in the H441 cell line

Abstract

Fetuses of mothers with uncontrolled gestational diabetes have an increased risk of developing neonatal respiratory distress syndrome and are frequently hyperinsulinemic, thus it has been proposed that high levels of insulin delay fetal lung maturation. We have shown previously that insulin inhibits the accumulation of mRNA for the surfactant-associated proteins A and B (SP-A and SP-B) in human fetal lung explants maintained in vitro. To test the hypothesis that the inhibitory effects of insulin on the surfactant proteins are the result of a direct action of insulin on the lung epithelial cell, we evaluated the effects of insulin in the H441 cell line, a human pulmonary adenocarcinoma cell line that expresses SP-A and SP-B mRNA. We observed that insulin treatment for 48 h decreased SP-A mRNA and protein levels in a concentration-dependent manner when compared to controls. The inhibitory effect of insulin on SP-A mRNA levels was apparent as early as after 4 h of exposure. SP-B mRNA levels were also significantly decreased by insulin in a concentration-dependent manner. Insulin, at 2.5 μg/ml, inhibited SP-A gene transcription by approx. 67%, and inhibited SP-B gene transcription by about 32%. There was no significant effect of insulin on SP-A or SP-B mRNA stability. Thus, we have observed a pattern of insulin inhibition of SP-A and SP-B gene expression in the H441 lung epithelial cell line similar to that previously observed in human fetal lung explants, which are comprised of both epithelial and mesenchymal cells. Our findings provide further evidence that insulin may delay fetal lung maturation by inhibiting SP-A and SP-B gene expression. Furthermore, our findings suggest that the inhibitory effects of insulin are, at least partially, the result of a direct action on the lung epithelial cell.