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Fig. 8 | Molecular Neurodegeneration

Fig. 8

From: Calcium-dependent cytosolic phospholipase A2 activation is implicated in neuroinflammation and oxidative stress associated with ApoE4

Fig. 8

Inhibition of cPLA2 reduces ApoE4 mediated up-regulation of LTB4, ROS, and iNOS levels. A, B and C ApoE3 and ApoE4 primary astrocytes from mice pups were pre-treated with medium (Ctrl) or the cPLA2 inhibitor (pyrrophenone) for 30 min followed by treatment with TNFα plus IFNγ together for 18 hours. A A representative blot showing total and phosphorylated-cPLA2 levels. The quantification was from 3 independent repeats. B LTB4 levels in the culture medium were measured by the assay kit. C A representative blot showing iNOS expression in cell lysate. The quantification was from 2 independent repeats. D ApoE3 and ApoE4 primary astrocyte were pre-treated with pyrrophenone for 30 min and then treated with medium or TNFα plus IFNγ together for 24 h. ROS levels were detected by the DCFDA probe. E, F ApoE4 primary astrocytes were transfected with cPLA2 siRNA or non-target (NT) siRNA for 48 hours and then treated with medium or TNFα plus IFNγ together for 24 hours. E cPLA2 protein levels in cell lysate were detected by WB. F LTB4 levels in the culture medium were measured by the assay kit. WB, Western Blot. DCFDA: 2′,7′–dichlorofluorescin diacetate. LTB4: leukotriene B4. Data are represented as mean ± SEM and analyzed by Student’s t-test (two-tailed). Two-way ANOVA was used in A, C, and D for group comparisons. *p < 0.05, **p < 0.01, ***p < 0.001

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