Protective effect of the flavonoid rutin against ferrous sulphate cytotoxicity to rat glioma C6 cells
Iron, Rutin, Cell Biology, Flavonoid, Ascorbic Acid, C6 Cells
INTRODUCTION: Oxidative stress is related to aging and several neurodegenerative disorders.
Flavonoids, on the other hand, have an important action in biological systems, for example as
scavenging free radicals, acting as antioxidant agents, either alone or together with molecules
such as ascorbic acid. Flavonoids like rutin stand out for their antioxidant and cytoprotective
capacity, therefore, they are promising in this field. AIM: To investigate the protective effect of
the flavonoid rutin against the cytotoxicity of ferrous sulfate in in vitro models of the C6 strain of
rat glioma, and to assess whether ascorbic acid interferes with this pharmacological activity.
METHODS: C6 cells were cultured under sterile conditions at 37ºC in an atmosphere with 5%
CO2 until reaching confluence. Specific modulations were made for each step; to determine the
minimum cytotoxic concentration of ferrous sulfate, the dilution occurred from 22 mM. The
ferrous sulfate cytotoxicity test was performed after 24h of treatment using the MTT test. For a
minimum protective concentration of rutin, a dilution was made (from 500 µM for 24h), followed
by treatment with ferrous sulfate (at 13 mM for 24h), as well as an analysis test of the protection
of rutin cells through the MTT test. To verify interference from ascorbic acid, an experiment was
carried out in which C6 cells were pretreated with rutin at the minimum protective concentration
(5 µM for 24 h), being then treated with ferrous sulfate (at 13.7 mM), varying the concentration
of ascorbic acid (between 6 - 300 µM per 24h of treatment) using the MTT test. RESULTS: The
minimum cytotoxic concentration of ferrous sulfate for C6 cells was 0.7 mM after 24 h. Rutin
exhibited a partial protective effect in cell viability tests. The minimum protective concentration
of rutin (5 µM) prevented a significant percentage of cell death induced by ferrous sulfate at 13
mM after 24 h. Ascorbic acid did not interfere with the protective effect of rutin.
CONCLUSIONS: Rutin has a cytoprotective action against oxidative damage from ferrous
sulfate, in C6 cells, ascorbic acid did not interfere in this protection.