Effect of gallic acid on Alkaline phosphatase gene expression in vascular smooth muscle cells

Background and purpose: Vascular calcification is an important factor in pathogenesis of atherosclerosis. Studies have shown that alkaline phosphatase increases vascular calcification. Here we investigated the effect of gallic acid on alkaline phosphatase gene expression in vascular smooth muscle cells (VSMCs). Materials and methods: In this experimental study humans aorta VSMCs were incubated with beta glycerol phosphate as calcification-inducing media. Then these cells were treated with 160, 180 and 200 µMol concentration of gallic acid for 24h, 48h and 72h. The total RNA was extracted and cDNA was synthesized and then alkaline phosphatase expression was measured by real time PCR. Alkaline phosphatase specific activity was measured by spectrophotometry. Results: Overall, 160, 180 and 200 µMol concentration of gallic acid decreased alkaline phosphatase gene expression in vascular smooth muscle cell by 1.98, 2.03, and 3.16 folds, respectively after 72h compared with the control group. The alkaline phosphatase specific activity also decreased compared to that of the control group. Conclusion: Our results showed that gallic acid decreased the expression and activity of alkaline phosphatase suggesting that this antioxidant compound may attenuate vascular calcification

Micro-CT imaging reveals<i> Mekk3 </i>heterozygosity prevents cerebral cavernous malformations in <i>Ccm2</i>-deficient mice

Mutations in CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation in humans. Mouse models of CCM disease have been established by deleting Ccm genes in postnatal animals. These mouse models provide invaluable tools to investigate molecular mechanism and therapeutic approaches for CCM disease. However, the full value of these animal models is limited by the lack of an accurate and quantitative method to assess lesion burden and progression. In the present study we have established a refined and detailed contrast enhanced X-ray micro-CT method to measure CCM lesion burden in mouse brains. As this study utilized a voxel dimension of 9.5μm (leading to a minimum feature size of approximately 25μm), it is therefore sufficient to measure CCM lesion volume and number globally and accurately, and provide high-resolution 3-D mapping of CCM lesions in mouse brains. Using this method, we found loss of Ccm1 or Ccm2 in neonatal endothelium confers CCM lesions in the mouse hindbrain with similar total volume and number. This quantitative approach also demonstrated a rescue of CCM lesions with simultaneous deletion of one allele of Mekk3. This method would enhance the value of the established mouse models to study the molecular basis and potential therapies for CCM and other cerebrovascular diseases