Corticosteroid effects on ventilator-induced diaphragm dysfunction in anesthetized rats depend on the dose administered

<p>Abstract</p> <p>Background</p> <p>High dose of corticosteroids has been previously shown to protect against controlled mechanical ventilation (CMV)-induced diaphragmatic dysfunction while inhibiting calpain activation. Because literature suggests that the calpain inhibiting effect of corticosteroid depends on the dose administered, we determined whether lower doses of corticosteroids would also provide protection of the diaphragm during CMV. This may be important for patients undergoing mechanical ventilation and receiving corticosteroids.</p> <p>Methods</p> <p>Rats were assigned to controls or to 24 hours of CMV while being treated at the start of mechanical ventilation with a single intramuscular administration of either saline, or 5 mg/kg (low MP) or 30 mg/kg (high MP) of methylprednisolone.</p> <p>Results</p> <p>Diaphragmatic force was decreased after CMV and this was exacerbated in the low MP group while high MP rescued this diaphragmatic dysfunction. Atrophy was more severe in the low MP group than after CMV while no atrophy was observed in the high MP group. A significant and similar increase in calpain activity was observed in both the low MP and CMV groups whereas the high dose prevented calpain activation. Expression of calpastatin, the endogenous inhibitor of calpain, was decreased in the CMV and low MP groups but its level was preserved to controls in the high MP group. Caspase-3 activity increased in all CMV groups but to a lesser extent in the low and high MP groups. The 20S proteasome activity was increased in CMV only.</p> <p>Conclusions</p> <p>Administration of 30 mg/kg methylprednisolone during CMV protected against CMV-induced diaphragm dysfunction while 5 mg/kg was more deleterious. The protective effect is due mainly to an inhibition of the calpain system through preservation of calpastatin levels and to a lesser extent to a caspase-3 inhibition.</p

Regulation of cytosolic calcium in skeletal muscle cells of the mdx mouse under conditions of stress.

1. In Duchenne muscular dystrophy (DMD) dysregulation of cytosolic calcium appears to be involved in the degeneration of skeletal muscle fibres. Therefore, we have studied the regulation of the free cytosolic calcium concentration ([Ca2+]c) under specific stress conditions in cultured myotubes isolated from the hind limbs of wild-type (C57BL10) and dystrophin-deficient mutant mdx mice. [Ca2+]c in the myotubes was estimated by the use of the Ca(2+)-sensitive fluorescent dye, fura-2. 2. Resting [Ca2+]c was similar in mdx and normal myotubes (35 +/- 9 nM and 38 +/- 11 nM, respectively). However, when mdx myotubes were exposed to a high extracellular calcium concentration ([Ca2+]c) of 40 mM, the [Ca2+]c was elevated to 84 +/- 29 nM, compared to 49 +/- 7 nM in normal myotubes. 3. Lowering the osmolarity of the superfusion solution from 300 mOsm to 100 mOsm resulted also in a rise in [Ca2+]c which was about two times higher for mdx (243 +/- 65 nM) than for C57BL10 (135 +/- 37 nM). Replacing extracellular Ca2+ by EGTA (0.2 mM) prevented the rise in [Ca2+]c in both mdx and normal myotubes when exposed to the low osmolarity solution. 4. Gadolinium ion (50 microM), an inhibitor of Ca2+ entry, antagonized the rise in [Ca2+]c of myotubes superfused with 40 mM [Ca2+]c by 20-40% for both mdx and C57BL10 cells, but did not significantly reduce the rise in [Ca2+]c when the cells were exposed to the hypo-osmotic buffer (100 mOsm). 5. Incubation of the cell culture for 3-5 days from the onset of induction of myotube formation with the membrane permeable protease inhibitor, calpeptin (50 microM) abolished the rise in [Ca2+]c in mdx myotubes upon exposure to hypo-osmotic shock. 6. Treatment of the cell culture for 3-5 days with alpha-methylprednisolone (PDN, 10 microM) attenuated the rise in [Ca2+]c following hypo-osmotic stress for both normal and mdx myotubes by about 50%. 7. The results described here suggest an increased permeability of mdx myotubes to Ca2+ under specific stress conditions. The ameliorating effect of PDN on [Ca2+]c could explain, at least partly, the beneficial effect of this drug on DMD patients

Excess recovery heat production by isolated muscles from mice overexpressing uncoupling protein-3

Contractile and energetic performance of bundles of muscle fibres from the soleus of mice overexpressing uncoupling protein 3 (UCP-3tg) were compared with the performance of bundles from wild-type mice. Force and heat production were measured during a series of thirty 0.2 s isometric tetani at Lo, the length optimal for force. UCP-3tg fibres were as strong as the wild-type and maintained force in the series equally well; in the first tetanus force was 116.9 ± 15.1 and 133.3 ± 19.7 mN mm−2 respectively (all values means ± s.e.m., n = 6 for UCP-3tg and n = 5 for wild-type). Heat production was partitioned into initial heat (due to contractile ATPases and the creatine kinase reaction) and recovery heat (due to other ATP-supplying processes) and expressed relative to the first cycle total heat. Initial heat production was similar for the UCP-3tg and wild-type fibres, decreasing during the series from 0.799 ± 0.052 to 0.661 ± 0.061 relative units (UCP-3tg), and from 0.806 ± 0.024 to 0.729 ± 0.039 relative units (wild-type). In both types the recovery heat was small at the start of the series and increased as the series progressed. At the end of the series, recovery heat production by UCP-3tg fibres, 1.575 ± 0.246 relative units, was twice that of the wild-type fibres, 0.729 ± 0.072 relative units. The extra recovery heat represents inefficient recovery in UCP-3tg fibres. This is the first direct evidence of enhanced energy dissipation as heat when UCP-3tg is overexpressed