Fig. 3

Activation of mGluR5 partially inhibited microglia activation induced by α-syn. a BV2 cells were transfected with myc-α-syn or vector for 24 h followed by CHPG (150 μM) administration for 24 h, followed by pre-treatment with CHPG for 1 h before LPS (100 ng/mL) treatment for 24 h, or followed by combination with these two applications. Immunoblotting analysis of cell lysates was detected by various antibodies. b–e Quantification of protein bands was normalized to β-tubulin (mGluR5, COX-2) and β-actin (α-syn, iNOS, TNF-α) (b–d). The p-p38 and p-NF-κB p65 levels were normalized to total p38 and NF-κB p65, respectively (e). f–h Culture supernatants from BV2 cells were assayed for NO (f), PGE₂ (g), and TNF-α (h) release by ELISA. Cells transfected with vector without drugs administration served as control (a–h). i Primary microglia were infected with LV-NC or LV-α-syn followed by CHPG (150 μM) administration for 24 h, followed by pre-treated with CHPG for 1 h before LPS (10 ng/mL) treatment for 24 h, or followed by combination with these two treatments. Cell lysates were probed by using indicated specific antibodies. j–m Quantification of protein bands was normalized to β-tubulin (mGluR5, COX-2) and β-actin (α-syn, iNOS, TNF-α) (j–l). The p-p38 and p-NF-κB p65 levels were normalized to total p38 and NF-κB p65, respectively (m). n–p Culture supernatants from primary microglia were assessed for NO (n), PGE₂ (o), and TNF-α (p) release by ELISA. Cells infected with LV-NC without drugs treatment served as control (i–p). Data shown in this figure represent mean ± SEM (n ≥ 3). The statistical significance was determined by one-way ANOVA followed by Dunnett’s test. ***p < 0.001 versus control (ctr); ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 versus α-synuclein (α-syn) transfection or the LV-α-syn infection. ^&p < 0.05 and ^&&&p < 0.001 versus LPS. ^$$p < 0.01 and ^$$$p < 0.001 versus the LPS accompanied by α-syn. ns, not significant