Role of mechanical stretching and lipopolysaccharide in early apoptosis and IL-8 of alveolar epithelial type II cells A549.

OBJECTIVE: To investigate the effects of mechanical stretching and lipopolysaccharide (LPS) on the early apoptosis and IL-8 production of alveolar epithelial type II cells A549. METHODS: The experimental matrix consisted of three integrated studies. In the first study, A549 cells were subjected to different stretching strain frequency and duration time to see the effects on the early apoptosis. In the second study, A549 cells were subjected to mechanical stretch (15% 4 h, 0.5 Hz) and LPS (1 or 100 ng/mL) to see whether mechanical strain and LPS also have an addictive effect on the early apoptosis. In the third study to investigate whether this addictive effect could be induced by LPS and mechanical stretch on IL-8 production, A549 cells were subjected to LPS (100 ng/mL) and mechanical strain (15%, 0.5 Hz, 4 h). Real time PCR and enzyme linked immunosorbent assay were used to measure mRNA and protein level of IL-8. The early apoptosis was detected by flow cytometry. RESULTS: Mechanical stretch induced the early apoptosis in a force and frequency and time-dependent manner. In the presence of LPS, mechanical stretch enhanced LPS-induced early apoptosis, especially in 100 ng/mL LPS group compared with 1 ng/mL LPS and the control group. Mechanical stretch increased IL-8 production and enhanced LPS-induced IL-8 screation both in mRNA and protein levels. CONCLUSIONS: Mechanical stretch can induce the early apoptosis and IL-8 secretion. Mechanical stretch and LPS have an addictive effect on the early apoptosis and IL-8 production in alveolar type 2 cells, which is one of the mechanisms of ventilator-induced lung injury.

Response of alveolar type II epithelial cells to mechanical stretch and lipopolysaccharide.

BACKGROUND: Although many therapeutic strategies have been developed clinically, the mortality associated with acute respiratory distress syndrome remains very high. OBJECTIVES: In this research, we used a cytomechanical method to elucidate the reason for this. METHODS: A549 cells were stimulated with lipopolysaccharide (LPS; 1 or 100 ng/ml) and/or mechanical stretch (5, 15, 30%) in varying frequency (0.2, 0.5, 1 Hz) at indicated time (1, 2, 4 h). Real time PCR and enzyme-linked immunosorbent assay were used to measure mRNA and protein levels of IL-8. RESULTS: In the presence of mechanical stretch, 100 ng/ml LPS significantly increased IL-8 production after 4 h of 5% stretch (p < 0.05). In the presence of LPS, stretch enhanced LPS-induced IL-8 protein production in a force-, time- and frequency-dependent manner. At both the 1- and 4-hour time points, mechanical stretch and LPS increased IL-8 mRNA levels, respectively, and stretch enhanced LPS-induced IL-8 mRNA levels (p < 0.05). CONCLUSIONS: Using cytomechanic methods, we found a synergistic effect of LPS and mechanical stretch on IL-8 production. The response of alveolar type II cells to mechanical stretch depends on their different pathologic states and the applied mechanical stretch, which may reversely influence the outcome of patients with acute respiratory distress syndrome.

Activations of mitogen-activated protein kinase and nuclear factor-kappaB by mechanical stretch result in ventilation-induced lung injury.

Mechanical ventilation is an important therapeutic technique for patients with respiratory failure. Nonetheless, it may cause or worsen lung injury. The specific triggers for cytokine release and the cellular origins of the inflammatory mediators in ventilation-induced lung injury (VILI) have yet to be defined. With the development of cytomechanics, we can study the lung cell response to mechanical strain. The initial step is mechanosensation, including stretch-activated ionchannels and the ECM-integrin-cytoskeleton pathway. Several intracellular signaling pathways then are activated and eventually result in increased transcription of specific genes. Mitogen-activated protein kinase cascade, nuclear factor(NF)-kappaB, PKC are all activated by mechanical stretch. But the mechanisms regulating lung stretch-induced cytokine production are still unclear. I hypotheses mechanical stretch initiate specific genes transcription, then the cytokines stimulate the cell again. This formed a positive feed back loop, which caused VILI. These studies may lead to the identification of new targets for therapeutic interventions and help to develop less aggressive ventilation strategies for patients with acute respiratory failure.