Glucose decreases steady state mRNA content of hydrophobic surfactant proteins B and C in fetal rat lung explants.

The streptozotocin-induced diabetic (STZ-DB) rat model is associated with fetal hyperglycemia, but with low to normal plasma insulin concentration. Because surfactant protein (SP) mRNA content in fetal rat lung is decreased in STZ-DB pregnancy, we investigated the effect of increasing concentrations of glucose on SP gene expression in lung organ cultures. SP mRNA content (SP-A, SP-B, SP-C) was assessed by Northern blot analysis in fetal day 20 lung explants (term = 22 days) cultured for 44 hours in medium containing 10, 25, 50, or 100 mM glucose. Our findings were (1) No consistent alteration in SP-A mRNA content was observed at different glucose concentrations (P > .05); (2) SP-B and SP-C mRNA content were reduced in a dose-dependent manner when glucose concentration was increased from 10 mM to 100 mM. The mRNA content, compared to 10 mM glucose, decreased to 50-60% at 25 mM glucose, to 20-25% at 50 mM glucose, and to lower than 10% at 100 mM glucose (P < .01). These findings indicate that the decrease in SP-B and SP-C mRNA in fetuses of STZ-DB rats may be, in part, due to a direct effect of hyperglycemia, whereas the decrease in SP-A mRNA content in STZ-DB rats appears to be due to other effects of diabetes in pregnancy.

Expression and cDNA sequence of helix destabilizing protein (HDP) in rat lung.

The current study was undertaken to investigate genes that show alteration in expression during late fetal rat lung development. Using the differential display (DD) method, an initially unknown transcript was found to be abundant on day 18 compared to day 22 of gestation. This finding was confirmed by RNA blot analysis. Subsequent screening of a rat lung cDNA library with the unknown DD cDNA as a probe revealed a clone that encodes helix destabilizing protein (HDP), similar to that reported in rat brain (J Biol Chem. 1986, 261:3536-3543). Although there were several differences between lung and brain cDNAs at the 3' untranslated region (UTR), the 5' end was well conserved. There was a gradual decrease in HDP mRNA content beyond day 18 of gestation, as lung maturation increased. Similarly, a lower content of HDP mRNA was observed in maternal heart and brain tissues than in fetal heart and brain tissues from day 18 of gestation. Maternal dexamethasone treatment that promotes lung maturation did not have an impact on HDP expression. Because of the much higher abundance of HDP mRNA during fetal organ development compared to adult tissues, we speculate that HDP plays an important role during lung development. A portion of this work was previously presented (FASEB J. 1996;10:A26).

Bronchodilator response in pulmonary disease at two different states of respiratory mechanics.

It would be convenient to be able to measure airway responsiveness to bronchodilator drugs with a sequential use of oscilloresistometry and spirometry, which may allow the comparison of the response to bronchodilator at two different states of respiratory mechanics i.e., resting tidal breathing at functional residual capacity (FRC) versus forced expiratory manoeuvre from total lung capacity to residual volume. The evaluation of airway resistance and forced expiratory volume at 1 s (FEV1) may thus assist in the interpretation of bronchial responsiveness tests (BRT) to pick up responders among non-responders to administered bronchodilator. Such a concept was verified in 54 patients with respiratory disease before and 10 min after inhalation of 200 micrograms of salbutamol. Within 10 min following salbutamol challenge the increase for forced vital capacity (FVC), FEV1 and peak expiratory flow rate (PEFR) was 23, 27 and 39% in 27 asthmatic patients of group A (p < 0.005). On the other hand, 27 patients with chronic obstructive pulmonary disease in group B did not show any significant change in FEV1 and PEFR (p > 0.05). However, they showed a decrease in airway resistance of 30% (p < 0.005) during normal tidal breathing at FRC. Twenty-two normal controls in group C showed values of airway resistance and expiratory flow rates to be within the normal range and the response to administered salbutamol inhalation was not significant (p > 0.05). It may be concluded that the airway responsiveness to bronchodilators using only FEV1 as an index of BRT should be interpreted with caution in non-responders because they may not be able to tolerate the mechanical challenge in the form of an FCV manoeuvre but show a significant decrease in airway resistance during resting tidal breathing at FRC.

Dexamethasone enhances surfactant protein gene expression in streptozotocin-induced immature rat lungs.

Because surfactant protein (SP) mRNA levels in rat fetuses are increased by maternal dexamethasone (dex) treatment and decreased in streptozotocin-induced diabetic (STZ-DB) pregnancy, we investigated the in vivo effects of dex on SP gene expression in STZ-DB pregnancy. The mRNA levels of SP (SP-A, SP-B, SP-C) were assessed in d 18 and 20 fetuses by Northern blot analysis, and nuclear run-on assays were performed with lung nuclei from d 20 fetuses (term = 22 d). Our findings indicate: 1) dex causes a greater increase in SP-A and SP-B mRNA levels in d 18 (12-16-fold) compared with day 20 (4-6-fold) fetuses (p < 0.05) in normal and STZ-DB pregnancy; 2) a 2-3-fold increase in SP-C mRNA levels was observed in response to dex in d 18 and 20 fetuses; 3) the increase in transcription of SP-A and SP-B in d 20 fetuses after dex is 68 and 60%, respectively, of the increase in their mRNA levels whereas in STZ-DB, the decrease in transcription compared with mRNA levels is 3.67-fold for SP-A and 2.42 fold SP-B; and 4) changes in SP-C transcription in either in vivo model, dex-treated or STZ-DB, correspond well with changes in mRNA levels. Together, these findings indicate that dex can enhance SP expression in STZ-DB immature lungs and support differential regulation of fetal SP genes in the models studied.