3.4. Effects of α-Klotho on GCM -induced cytotoxicity of mouse
primary neuronal culture
Primary glia culture was pretreated with serum-free medium (GCM) or with
1nM α-Klotho (GCM-KL) for 24 hours, and then challenged with 1 µg/mL LPS
for 24 hours. Initially, it was necessary to determine the amount of GCM
coming from LPS-challenged glia cell (GCM group) which caused neuronal
death. Thus, we replaced 10, 25, and 50% of the cultured medium of the
neuronal culture by GCM. The results showed that when the concentration
of 25 and 50% of the neuronal culture medium were switched to GCM,
there was an increase in neuronal toxicity when compared to the control
group (Fig.5).
After determining the concentration of GCM that induced neurotoxicity,
we investigated whether the GCM from α-Klotho pretreated glial culture
(1nM) and stimulated with LPS (GCM-KL) would be able to decrease the
neuronal death when compared to GCM at both 25 and 50% concentrations
(Fig.6). The results showed that 25% of GCM-KL reversed the increase in
neuronal death caused by GCM (Fig.6A), but α-Klotho pretreatment was not
able to induce any effect to a high concentration (50% of GCM-KL)
(Fig.6B).