Effect of metabolic acidosis on the growth hormone/IGF-I endocrine axis in skeletal growth centers.

BACKGROUND

Chronic metabolic acidosis (CMA) adversely affects bone metabolism and skeletal growth. Given the cardinal role played by the local growth hormone (GH)/insulin-like growth factor-I (IGF-I) in promoting cell proliferation and differentiation in growth plates, we tested the effect of CMA on the GH/IGF-I axis in a skeletal growth center.

METHODS

We employed an in vitro organ culture system using the murine mandibular condyle as a model for endochondral active growth center. Condyles from six-day-old ICR mice were cultured in BGJb medium of either neutral pH (pH approximately 7.4) or acidic pH (pH approximately 7.15). After 24, 48, 72, and 96 hours of culture, the condyles were washed, fixed in formaldehyde, and processed for paraffin embedding. We assessed histologic markers of the growth center. In addition, the protein level and mRNA expression for the different components of the GH/IGF-I axis were evaluated by immunohistochemistry and in situ hybridization, respectively. Finally, we evaluated the effect of acidosis on the biological functions mediated by GH and IGF-I (namely, proliferation and differentiation of cartilage cells in the active growth center).

RESULTS

Following three to four days in acidic conditions, there was a marked reduction in the size of young chondrocytic population, suggesting a defect in the process of endochondral differentiation. Immunohistochemistry and in situ hybridization analyses revealed a marked reduction in the expression of the IGF-I receptor, as well as in the GH receptor. These changes were already evident after 48 hours of incubation in acidic conditions. At 48 hours of acidosis, there was also a marked reduction in the expression of IGF-I both under basal conditions (nonstimulated) and following stimulation with GH. The expression of IGF binding protein 2 (IGFBP-2) and IGFBP-4, which serve as negative modulators of IGF-I, was enhanced in CMA. IGF-I markedly stimulated chondrocytic proliferation (assessed by BrdU incorporation into DNA) and differentiation (assessed as cartilage specific proteoglycan expression). These responses were markedly attenuated in acidic conditions.

CONCLUSION

CMA exerts an anti-anabolic effect in bone growth centers, which is partly related to a state of resistance to GH and IGF-I, created by CMA. This phenomenon may underlie the disturbance in longitudinal bone growth in CMA (that is, renal tubular acidosis) and may contribute to renal osteodystrophy in patients suffering from chronic renal failure.