Zingiber officinale Roscoe Aqueous Extract Modulates Matrixmetalloproteinases and Tissue Inhibitors of Metalloproteinases Expressions in Dengue Virus-infected Cells: Implications for Prevention of Vascular Permeability

Purpose: To investigate the effect of the aqueous extract of Zingiber officinale Roscoe. (ZOA) rhizome on the activity and expression of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 using an in vitro model of Dengue virus (DV) infection. Methods: Z. officinale rhizomes were extracted with water by continuous shaking for 5 days. The total phenolic content in extract was measured by Folin-Ciocalteu method. High performance liquid chromatography (HPLC) was employed to define qualitative and quantitative content of [6]-gingerol in ZOA. The median inhibitory concentration (IC50) value of ZOA for Vero cells was determined by 3-(4,5 dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To induce MMPs production, Vero cells were infected with DV3. The modulatory effect of ZOA on the activity and expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed using gelatin zymography and quantitative Real-time polymerase chain reaction (RTPCR), respectively. Results: The yield of the ZOA was 7.98%. Total phenolics in ZOA was 68.17 ± 0.28 mg GAE/g of extract and it contained 29.32 ± 1.97 mg 6-gingerol/g of extract.The half-maximal inhibition concentration (IC50) of ZOA was 348.8 μg/mL for Vero cells. DV infection of Vero cells significantly elevated the production of soluble gelatinolytic MMP-2 and to a lesser extent, MMP-9, and their activities were significantly inhibited by ZOA in a dose-dependent manner. A significant down-regulation of MMP-2, MMP-9 mRNA expression and up-regulation of TIMP-1, TIMP-2 mRNA expression were observed in DV-infected Vero cells following treatment with ZOA, and it occurred in a dose-dependent manner. Conclusion: The findings of this study suggest that ZOA may ameliorate plasma leakage in dengue virus infection and decrease the chances of severe dengue complications, dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) by inhibiting the activities and expression of MMP-2 and MMP-9 while upregulating the expression of TIMP-1 and TIMP-2.