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Carlos Fernández-Moriano et al.

350? # + H2O2
300? Ps? Ug? Trolox?
250? ##
200? # 0.01?mM?
150?
100?

50?
0?
Control? Hydrogen? Cb?
peroxide?
0.1?mM?

Figure 4. Effects of pretreatments with Cb, Ps, Ug and trolox on H2O2-induced caspase-3 over-activation
(concentrations: 0.5 µg/mL for Cb and Ug and 0.25 µg/mL for Ps; 24 h). Cells were treated with lichen
extracts and H2O2 (0.1mM; 30 mins). Means ± SD, # p < 0.05 V s H2O2.

3.3.d. Gluthatione levels the other hand, pretreatments with the three lichen extracts,
at the concentrations previously mentioned for each one,
GSH and GSSG levels were measured as important ameliorated antioxidant capacity in neurons by increasing
oxidative stress markers by using fluorometric methods. It GSH/GSSG ratio compared to cells treated only with
was evidenced that glutathione is mostly found as its H2O2. Methanol extract of Usnea ghattensis was the most
reduced form in control cells and, when SH-SY5Y cells effective in restoring normal GSH and GSSG levels
were incubated with H2O2 (0.1 mM for 30 min), GSH against the oxidative damage induced by H2O2 and the
levels decreased significantly (7.77 nmol/mg protein) only one showing a statistically significant difference.
when compared with control cells (9.47 nmol/mg protein), Results obtained for all extracts and trolox (as reference
while the levels of the oxidized form also increased antioxidant) are expressed in Figure 5.
significantly (2.62 nmol/ mg protein for control cells and
9.05 nmol/mg protein for cells treated only with H2O2). On

000000000000000000425214313?????????
000?

Figure 5. Effects of pretreatments with Cb, Ps, Ug and trolox on H2O2-induced changes in
GSH/GSSG ratio (concentrations: 0.5 µg/mL for Cb and Ug and 0.25 µg/mL for Ps; 24 h). Cells were
treated with lichen extracts and H2O2 (1mM). Means ± SD, # p < 0.05 Vs H2O2.

172 @Real Academia Nacional de Farmacia. Spain