Effect of Ulinastatin on the Expression of iNOS, MMP-2, and MMP-3 in Degenerated Nucleus Pulposus Cells of Rabbits

Abstract

We examined the effects of ulinastatin on the expression of inducible nitric oxide synthase (iNOS), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-3 (MMP-3) in degenerated nucleus pulposus (NP) cells of rabbits induced by interleukin (IL)-1β in vitro. An in vitro NP cell culture model was set up with enzyme digestion. NP cells from adult white rabbits were divided into six groups: the normal control group, the ulinastatin control group (320 U/mL ulinastatin), the induced group (10 ng/mL IL-1β), and three inhibition groups (IL-1β followed by 160, 320, or 640 U/mL ulinastatin). After a 2-day culture, the NP cells were collected for immunohistochemical staining for MMP-2 and MMP-3 and spectrophotometric analysis of the amount of iNOS. Immunohistochemical staining showed that the expression of MMP-2 and MMP-3 proteins in NP cells decreased in the inhibition groups compared with the induced group, which was in inverse proportion to the ulinastatin concentration. Spectrophotometric results showed that, compared with the induced group, the iNOS content in each inhibition group decreased, most significantly in the 320 U/mL group. Ulinastatin effectively inhibited the increased expression of MMP-2, MMP-3, and iNOS in degenerated NP cells induced by IL-1β in vitro. It suggests that ulinastatin may potentially be useful for clinical therapy of intervertebral disc degeneration.

• nucleus pulposus
• trypsin inhibitor
• matrix metalloproteinase-2 and matrix metalloproteinase-3
• interleukin-1β
• nitric oxide synthase