| Objective:Our aim was to explore the effects of ozone with different concentrations on the tissue structures and the expression levels of NO and IL-6 of HNP patients’nucleus pulposus.Methods:Nucleus pulposus specimens were obtained from sixteen HNP patients. Each specimen was randomly divided into five groups:normal group (group C) and group O2, O320, O340 and O360 (intervened by oxygen,20μg/ml ozone,40μg/ml ozone and 60μg/ml ozone respectively). Then cultured them in vitro for 72 hours, after which HE staining were performed to observe the tissue structure and qRT-PCR were used to determine the expression levels of iNOS mRNA and IL-6 mRNA. Media were collected for assays of NO (Griess reaction) and IL-6 (enzyme-linked immunosorbent assay).Results:1. Histological analysis:Diffused distribution cartilage cells and abundant and zonal distribution matrix could be seen in group C and group O2. While there were proliferated fibroblasts and reduced matrix in group O320 and group O340. And in group O360 it presented as hyperplastic collagen fibers and decreased and disorganized matrix.2. NO and IL-6 expression levels:Compared with group C, there were rised expression leverls of NO and IL-6 in group O320 and O340, to the contrary, those of group O360 could be seen a decline. No statistical differences were found between group C and group O2.3. qRT-PCR results:Expression levels of iNOS mRNA and IL-6 mRAN decreased in group O320 and O340 compared with group C, while expression levels of group O360 increased. But differences between group C and group O2 was not statistically significant.Conclusion:Ozone has the function of matrix degradation and collagen deposition.20μg/ml and 40μg/ml ozone can inhibit the expression levels of iNOS mRNA and IL-6 mRNA of HNP patients’ nucleus pulposus and decrease NO and IL-6 synthesis, while those expression levels increased when intervened by 60μg/ml ozone. |
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