Influence of rhDNAse on the duration of mechanical ventilation in intensive care patients - interim analysis of the LUFIT trial
European Journal of Anaesthesiology
Objective To examine the effects of short-term cyclic stretch on apoptosis in alveolar type II cells (A549). To study in vitro the direct influence of alveolar type II cells on mechanical stretch. Methods A549 were treated with different doses of lipopolysaccharide (LPS), 0 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1000 ng/ml, and then A549 were lengthened 5%, 15%, 30% using a FLEXCELL tension unit 4000, a vacuum-driven device that applies strain to cells, which were cultured in six-well plates
... ix-well plates coated with collagen-I, and 12 cycles/min for 4 hours. Apoptosis was measured using the flow cytometry method that measures annexin V and propidium iodide (PI) staining. The morphological changes of apoptotic cells were observed by transmission electron microscope. Results Apoptosis could be induced in alveolar type II cells (A549) by mechanical stretch. The percentage of annexin V + PI cells increased after being treated with cyclic stretch for 4 hours by 5%, 15%, 30% in all groups. The morphological features of apoptotic cells demonstrated by transmission electron microscope were as follows: shrinkage of the cell, chromatin condensation and aggregation under the nuclear membrane as a crescent or lump, membrane-encapsulated nuclear fragment or cell organ formed by invagination of the cell membrane, and apoptotic body formation followed by vacuolization. Conclusion Apoptosis induced by mechanical stretch and LPS is dose dependent. Mechanical stretch aggravates apoptosis especially in cells treated with LPS. Annexin V and PI double staining is a specific, sensitive, and quantitative method for analyzing apoptotic cells. It is also helpful to clarify the protective mechanism of low-volume ventilation in ARDS. PaO 2 /FiO 2 430 [421; 440] # 380 [349; 397] 165 [68; 289] # C (ml/cmH 2 O) 28 [24; 32]* 18 [16; 21]* 12 [8; 17]* R i (cmH 2 O/l/s) 4.1 [3.9; 4.5] 4.5 [4.3; 5.1] 5.1 [3.7; 7.9] # P < 0.05 control vs 24-hour peritonitis, *P < 0.05 control vs 12-hour and 24-hour peritonitis. Conclusions We conclude that an ALI/ARDS-like state is developed by most pigs during fecal peritonitis and that this peritonitis model may therefore serve as an extrapulmonary ARDS model. However, this condition develops after a prolonged period of approximately 12-18 hours, and the severity of the condition in single animals may be less predictable when compared with ARDS models induced by direct lung injury. Furthermore, it should be emphasized that pulmonary function in pigs is markedly different from humans in as much as no collateral ventilation exists in this species , and that pulmonary blood flow regulation is far more susceptible to hypoxia in pigs when compared with other species including humans . Lung function data derived from pig models should therefore always be interpreted cautiously if clinically relevant conclusions have to be drawn. Acknowledgement M Matejovic was supported by a grant from the Alexander von Humboldt Stiftung.