Effect of a nonprotein bioactive agent on the reduction of cyclooxygenase-2 and tumor necrosis factor–α in human intervertebral disc cells in vitro

Object Neurotropin is a nonprotein extract from the inflamed skin of rabbits inoculated with vaccinia virus. In the present study the authors sought to clarify the focal antiinflammatory effects of Neurotropin in intervertebral disc cells, and these effects were compared with those induced by the selective cyclooxygenase (COX)–2 inhibitor 6-methoxy-2-naphthylacetic acid (nabumetone). Methods Six human intervertebral disc specimens were harvested during spinal surgery for lumbar disc herniation. Cells were stimulated with 500 pg/ml of interleukin (IL)–1β in the presence of various concentrations of Neurotropin (0, 10−5, 10−4, and 10−3 Neurotropin Units/ml) or 50 μg/ml of nabumetone for 3 hours. The mRNA was extracted for polymerase chain reaction (PCR), and real-time PCR was used to quantify the mRNA levels of COX- 2, tumor necrosis factor (TNF)–α, and phospholipase A2. Cyclooxygenase-2, TNFα, and prostaglandin E2 (PGE2) protein concentrations were each determined by enzyme-linked immunosorbent assay. Resu...