Research Article
The Effects of Alpha-Linolenic Acid on the Secretory Activity of Astrocytes and β Amyloid-Associated Neurodegeneration in Differentiated SH-SY5Y Cells: Alpha-Linolenic Acid Protects the SH-SY5Y cells against β Amyloid Toxicity
Figure 12
The CM and ALA-CM pretreatment reduced acidic vesicular organelles (AVOs) production in the SH-SY5Y cells treated with Amyloid β (Aβ1-42). Representative fluorescence microscopy images of acridine orange (AO) staining (a) and the ratio of red to green fluorescence intensity ratio (R/GFIR) (b) were used to measure the late state of autophagy, which is characterized by AVOs production. On the day 6th, the SH-SY5Y cells (differentiated) were pretreated for 1 h with CM or ALA-CM before the addition of 5 μM Aβ1-42 for the next 24 h. The SH-SY5Y cells were also exposed to the cotreatment of CM and ALA-CM with Insulin Degrading Enzyme (IDE) to check whether insulin and IGF-I presence in CM and ALA-CM was responsible for the neuroprotective effect. The positive control was the SH-SY5Y cells treated with carbonyl-cyano-m-chlorophenylhydrazone-CCCP (10 μM). Next, the SH-SY5Y cells were subjected to vital staining with AO (see Materials and Methods section). Red to green fluorescence intensity ratio (R/GFIR) was calculated in at least 10 replicates for each treatment and nontreated controls. One-way ANOVA followed by Tukey’s multiple comparisons test at the 0.05 level was used to determine differences between the treated cells and untreated control cells. Results are presented as . Statistical differences between the treated cells and untreated control cells are indicated by asterisks ( for ; for ; for ; # versus the control group).
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