Crocetin Alleviates Inflammation in MPTP-Induced Parkinson’s Disease Models through Improving Mitochondrial Functions

Figure 5

Crocetin blocks interaction between ANT and Cyp D to inhibit mPTP opening. (a) The curve is a typical sample of mitochondrial swelling induced by 100 μM CaCl₂. Mitochondrial swelling was measured by monitoring mitochondrial size at an absorbance of 540 nm. 5 μM and 10 μM final concentrations of crocetin were added to the mitochondrial suspension for 5 min before adding CaCl₂. A change of absorbance within 20 min with a spectrophotometer indicated mitochondrial swelling. (b) Isolated mitochondria were pretreated with crocetin for 5 min followed by 100 μM CaCl₂ for 20 min, and then mitochondria were lysed and anti-VDAC antibody was added to pull down ANT and Cyp D. The levels of ANT and Cyp D pulled down by anti-VDAC were measured using immunoblotting. (c) BV2 cells were treated with 0.5 mM MPP+ for 6 h, and then 2.5 μM, 5 μM, and 10 μM crocetin were added and incubated for another 24 h. The ANT and Cyp D pulled down by VDAC were detected using immunoblotting. Data are expressed as mean ± standard deviation, and statistical analysis was performed using a one-way analysis of variance followed by Tukey’s post hoc test. versus control; , versus calcium group.

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