Controversy exists whether high frequency oscillatory ventilation with an active expiratory phase (HFO-A) should be used at low ventilator pressures or high alveolar volumes to minimize lung injury in the atelectasis-prone lung. We therefore ventilated 20 anesthetized, tracheostomized rabbits made surfactant-deficient by lung lavage in 1 of 3 ways: HFO-A at a high lung volume (HFO-A/HI), HFO-A at a low lung volume (HFO-A/LO), or conventional mechanical ventilation (CMV); all received 100% oxygen for 7 h. We examined oxygenation, lung mechanics, and lung pathology. Arterial oxygenation in the HFO-A/HI rabbits was kept greater than 350 mm Hg. Mean lung volume above FRC in these animals was 23.4 ml/kg. In rabbits ventilated with HFO-A/LO and CMV, arterial oxygen tensions were 70 to 100 mm Hg. Mean lung volumes were 7.8 and 4.3 ml/kg, respectively. Total respiratory system pressure-volume curves (P-V curves) showed no change from baseline in the HFO-A/HI group after 7 h of ventilation. The low lung volume groups (HFO-A/LO and CMV) showed a diminution in hysteresis of their P-V curves, lower total respiratory system compliance, more hyaline membranes and severe airway epithelial damage. (All changes significant with p less than 0.05). We conclude that maintenance of alveolar volume is a key mechanism in the prevention of lung injury during mechanical ventilation of the atelectasis-prone lung. For optimal outcome using high frequency oscillatory ventilation, alveoli must be actively reexpanded and then kept expanded using appropriate mean airway pressures.

Vinyl carbamate (VC) and ethyl carbamate (EC) induce the formation of lung tumors. The mechanism involves a two-step oxidation of EC to VC and VC to an epoxide, both of which are mediated mainly by CYP2E1. Interaction of the epoxide with DNA leads to the formation of DNA adducts, including 1,N(6)ethenodeoxyadenosine and 1,N(4)-ethenodeoxycytidine. The production of DNA adducts correlated with capacities for the bioactivation of VC, which are higher in the lungs of A/J than in C57BL/6 mice. Importantly, CYP2E1 is higher in the lungs of A/J than in C57BL/6 mice. Studies using F(1) (Big Blue x A/J) transgenic mice revealed the formation of mutations in the lambda cII gene after treatment with VC. Mutations induced by VC were mainly A:T-->G:C transitions and A:T-->T:A transversions, while mutations induced by EC were mainly G:C-->A:T transitions. An EC dose that was 17-fold higher than that for VC was required to produce a similar level of mutant frequency in the lung. Pretreatment of mice with the CYP2E1 inhibitor, diallyl sulfone, significantly inhibited the mutant frequencies induced by VC. Mutations in the endogeneous Kras2 gene were found in codon 61 of exon 2 and were identified as A:T transversions and A-->G transitions in the second base and A-->T transversions in the third base. These mutations were reduced by treatment of mice with diallyl sulfone before VC and coincided with a reduction in the number of lung tumors with Kras2 mutations. These findings affirmed that the metabolism of EC and VC is a prerequisite for, or at least substantially contributes to, initiation of the cascade of events leading to lung tumor formation.

1,1-Dichloroethylene (DCE) exposure evokes lung toxicity with selective damage to bronchiolar Clara cells. Recent in vitro studies have implicated CYP2E1 and CYP2F2 in the bioactivation of DCE to 2-S-glutathionyl acetate [C], a putative conjugate of DCE epoxide with glutathione. An objective of this study was to test the hypothesis that bioactivation of DCE is catalyzed by both CYP2E1 and CYP2F2 in murine lung. Western blot analysis of lung microsomal proteins from DCE-treated CD-1 mice showed time-dependent loss of immunodetectable CYP2F2 and CYP2E1 protein. Dose-dependent formation of conjugate [C] was observed in the lungs of CD-1 mice treated with DCE (75-225 mg/kg), but it was not detected after pretreatment with 5-phenyl-1-pentyne (5-PIP). Treatment of mice with 5-PIP and also with diallyl sulfone (DASO 2 ) significantly inhibited hydroxylation of p-nitrophenol (PNP) and chlorzoxazone (CHZX). Incubation of recombinant CYP2F3 (a surrogate for CYP2F2) and recombinant CYP2E1 with PNP and CHZX confirmed that they are substrates for both of the recombinant enzymes. Incubation of the recombinant enzymes with DASO 2 or 5-PIP significantly inhibited hydroxylation of both PNP and CHZX. Bronchiolar injury was elicited in CD-1 mice treated with DCE (75 mg/kg), but it was abrogated with 5-PIP pretreatment. Bronchiolar toxicity also was manifested in the lungs of CYP2E1-null and wild-type mice treated with DCE (75 mg/kg), but protection ensued after pretreatment with 5-PIP or DASO 2 . These results showed that bioactivation of DCE in murine lung occurred via the catalytic activities of both CYP2E1 and CYP2F2 and that bioactivation by these enzymes mediated the lung toxicity.

The purpose of this study was to investigate the expression and distribution of pulmonary CYP2E1 in mice. The CYP2E1 protein and mRNA were identified by immunoblotting and northern blotting, respectively, while the distribution of the CYP2E1 protein and mRNA was examined by immunohistochemistry and in situ hybridization, respectively. Protein immunoblotting revealed a single band of approximately M(r) 51,000 in lung microsomes of CD-1 male mice. Northern blotting with a 32P-labeled RNA probe for CYP2E1 detected a single species of approximately 2 kb that was similar in size to that of liver CYP2E1. Immunohistochemical studies with the avidin-biotin complex procedure showed that CYP2E1 was localized prominently in the nonciliated Clara cells but was not detected in the ciliated cells of the bronchiolar epithelium. In the lung parenchyma, immunoreactivity for CYP2E1 was evident at minimal levels in alveolar type II cells. In situ hybridization experiments with a 33P-labeled RNA probe showed that the CYP2E1 mRNA was also predominantly localized in the bronchiolar epithelium and was most prominent in the Clara cells. As was found for the CYP2E1 protein, the CYP2E1 mRNA was minimal in cells of the lung parenchyma. These results demonstrated that the CYP2E1 enzyme is preferentially expressed in Clara cells of murine lung. The concentration of CYP2E1 mainly in this cell population may be an important determinant underlying its susceptibility to cytotoxicities induced by xenobiotics bioactivated by this P450 isozyme.