Assessment of IL-6, IL-8 and TNF-α levels in the gingival tissue of patients with periodontitis.

In periodontal diseases, inflammatory mediators, including interleukin (IL)-6, IL-8 and tumor necrosis factor-α (TNF-α), may promote the degeneration of inflamed periodontal tissues. In previous studies, levels of these three cytokines were demonstrated to be elevated in inflammatory gingival tissues and gingival crevicular fluid. The aim of the present study was to quantify IL-6, IL-8 and TNF-α levels in the human gingival tissues of patients with periodontitis and to assess the correlation of these three cytokines with each other. In this study, human gingival tissues from 19 patients with periodontitis (male, n=14; female, n=5) were collected. The tissues were homogenized, centrifuged and the protein in the supernatant was quantified. Enzyme-linked immunosorbent assay (ELISA) was used in the measurement of the IL-6, IL-8 and TNF-α levels, and the mean levels were observed to be 8.41±0.25, 34.01±1.09 and 20.70±0.31 pg/ml, respectively. The mean levels of IL-8 were higher than those of the other two cytokines. In each sample, the level of TNF-α expression was consistently high, with little difference between the results, which contrasted with the fluctuations in IL-6 and IL-8 levels. The expression of the two ILs (IL-6 and IL-8) showed a positive correlation (r=0.932, P=0.01), whereas TNF-α levels were not correlated with IL-6 or IL-8 levels. These results suggest that IL-6, IL-8 and TNF-α may be relevant in the pathophysiology of periodontitis, and the measurement of these cytokines may be beneficial in the identification of patients with periodontitis.

Correlation of expression and activity of matrix metalloproteinase-9 and -2 in human gingival cells of periodontitis patients.

PURPOSE: Matrix metalloproteinases (MMPs) are capable of degrading extracellular matrix, and they are inducible enzymes depending on an inflammatory environment such as periodontitis and bacterial infection in periodontal tissue. Gingival inflammation has been postulated to be correlated with the production of MMP-2 and MMP-9. The objective of this study was to quantify the expression and activity of MMP-9 and -2, and to determine the correlation between activity and expression of these MMPs in human gingival tissues with periodontitis. METHODS: The gingival tissues of 13 patients were homogenized in 500 µL of phosphate buffered saline with a protease inhibitor cocktail. The expression and activity of MMP-2 and -9 were measured by enzyme-linked immunosorbent assay and Western blot analysis, and quantified by a densitometer. For the correlation line, statistical analysis was performed using the Systat software package. RESULTS: MMP-9 was highly expressed in all gingival tissue samples, whereas MMP-2 was underexpressed compared with MMP-9. MMP-9 activity increased together with the MMP-9 expression level, with a positive correlation (r=0.793, P=0.01). The correlation was not observed in MMP-2. CONCLUSIONS: The expression of MMP-2 and -9 might contribute to periodontal physiological and pathological processes, and the degree of MMP-9 expression and activity are predictive indicators relevant to the progression of periodontitis.

Inhibition of Hypoxia Inducible Factor Alpha and Astrocyte-Elevated Gene-1 Mediates Cryptotanshinone Exerted Antitumor Activity in Hypoxic PC-3 Cells.

Although cryptotanshinone (CT) was known to exert antitumor activity in several cancers, its molecular mechanism under hypoxia still remains unclear. Here, the roles of AEG-1 and HIF-1α in CT-induced antitumor activity were investigated in hypoxic PC-3 cells. CT exerted cytotoxicity against prostate cancer cells and suppressed HIF-1α accumulation and AEG-1 expression in hypoxic PC-3 cells. Also, AEG-1 was overexpressed in prostate cancer cells. Interestingly, HIF-1α siRNA transfection enhanced the cleavages of caspase-9,3, and PAPR and decreased expression of Bcl-2 and AEG1 induced by CT in hypoxic PC-3 cells. Of note, DMOG enhanced the stability of AEG-1 and HIF-1α during hypoxia. Additionally, CT significantly reduced cellular level of VEGF in PC-3 cells and disturbed tube formation of HUVECs. Consistently, ChIP assay revealed that CT inhibited the binding of HIF-1α to VEGF promoter. Furthermore, CT at 10 mg/kg suppressed the growth of PC-3 cells in BALB/c athymic nude mice by 46.4% compared to untreated control. Consistently, immunohistochemistry revealed decreased expression of Ki-67, CD34, VEGF, carbonic anhydrase IX, and AEG-1 indices in CT-treated group compared to untreated control. Overall, our findings suggest that CT exerts antitumor activity via inhibition of HIF-1α, AEG1, and VEGF as a potent chemotherapeutic agent.