Novel insights of the mechanism in uptake of modified-LDL in macrophage through phospholipase A2

• Project/Area Number: 24790749
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Circulatory organs internal medicine
• Research Institution: University of Yamanashi
• Principal Investigator: FUJIOKA Daisuke 山梨大学, 総合研究部, 助教 (70377513)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ホスホリパーゼA2 / 酸化LDL / Src / マクロファージ / ノックダウン / 過剰発現 / endosome

Outline of Final Research Achievements

We established a stable cell line which expresses low levels of group V sPLA2 with the use of shRNA-knockdown method in Raw264.7 mouse macrophage cell line. This cell line showed delayed- and decreased-activities in uptake and degradation of oxidized-LDL, and these cells also showed decreases of the level of actin polymerization and the activity of Rho GTPase which regulate the uptake of oxidized-LDL into the cells, in comparison with wild type cell line. The expression level and the enzymatic activity of Src were reduced in group V sPLA2-knockdown cell line, and these phenomenons suggested that group V sPLA2 interacts with the promoter region in Src and regulates the expression of Src.