The Synergistic Effects of Heat Shock Protein 70 and Ginsenoside Rg1 against Tert-Butyl Hydroperoxide Damage Model In Vitro

Figure 7

Effects of cotreatment on the damage cortex neurons. Neurons were grouped as C: media control; T: 10 μmol/L t-BHP; G: treated with G-Rg1 which dissolved into 10 μmol/L t-BHP; N: cocultured with NSCs in 10 μmol/L t-BHP; NG: cocultured with NSCs and treated with G-Rg1 which dissolved into 10 μmol/L t-BHP; NH: cocultured with overexpression of HSP70 NSCs in 10 μmol/L t-BHP NHG: cocultured with overexpression of HSP70 NSCs and G-Rg1 which dissolved in 10 μmol/L t-BHP. (a) Morphological changes of neurons, such as spines, filaments, and synapsis were pictured under SEM in C, T, G, N, NG, NH, and NHG at Day 3. (b) Western blot was performed to detect cleaved-Caspase-3 expression. Statistical comparisons were carried out with Student’s -test. Mean ± SEM. versus C group, versus T group, and versus monotherapy group. . (c) Representative images of immunofluorescence were shown at 200x magnification to demonstrate the MAP-2 positive cells; neurite lengths and number of neurites of each neuron were measured by Imaris software. (d), (e) The value of neurite lengths and number of neuritis were expressed as the mean ± SEM. Statistical comparisons were carried out with Student’s -test. versus C group, versus C group, and versus monotherapy group. . (f) Western blot was performed to detect NeuN and GFAP expression. (g), (h) The ratios of NeuN/β-actin and GFAP/β-actin were compared with C which had been standardized to 1. Statistical comparisons were carried out with Student’s -test. Values were presented as mean ± SEM. versus C group, versus T group, and versus monotherapy group (T, G, N, NG, and NH). . (i) Representative images of immunofluorescence were shown at 400x magnification to demonstrate the synaptic connection which were marked with antibody synaptophysin. (j), (k), and (l) Western blot was performed to detect HSP70, synaptophysin, and ChAT expression. Statistical comparisons were carried out with Student’s -test. Mean ± SEM. versus C group, versus T group, and versus monotherapy group. .

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