Neuroprotective action of alpha-Klotho against LPS-activated glia conditioned medium in primary neuronal culture

Author(s)

Nakao, Vinicius Wanatable ; Yokowama Mazucanti, Caio Henrique ; Lima, Larissa de Sa ; de Mello, Paloma Segura ; de Souza Port's, Natacha Medeiros ; Kinoshita, Paula Fernanda ; Leite, Jacqueline Alves ; Kawamoto, Elisa Mitiko ; Scavone, Cristoforo

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 12, n. 1, p. 12-pg., 2022-11-07.

Abstract

The alpha-Klotho is an anti-aging protein that, when overexpressed, extends the life span in humans and mice. It has an anti-inflammatory and protective action on renal cells by inhibiting NF-kappa B activation and production of inflammatory cytokines in response to TNF-alpha. Furthermore, studies have shown the neuroprotective effect of alpha-Klotho against neuroinflammation on different conditions, such as aging, animal models of neurodegenerative diseases, and ischemic brain injury. This work aimed to evaluate the effects of alpha-Klotho protein on primary glial cell culture against the proinflammatory challenge with LPS and how this could interfere with neuronal health. Cortical mixed glial cells and purified astrocytes were pretreated with alpha- alpha-Klotho and stimulated with LPS followed by TNF alpha, IL-1 beta, IL-6, IFN-gamma levels, and NF-kappa B activity analysis. Conditioned medium from cortical mixed glia culture treated with LPS (glia conditioned medium (GCM) was used to induce neuronal death of primary cortical neuronal culture and evaluate if GCM-KL (medium from glia culture pretreated alpha-Klotho followed by LPS stimulation) or GCM + LPS in the presence of KL can reverse the effect. LPS treatment in glial cells induced an increase in proinflammatory mediators such as TNF-alpha, IL-1 beta, IL-6, and IFN-gamma, and activation of astrocyte NF-kappa B. GCM treated-cortical neuronal culture induced a concentration-dependent neuronal death. Pretreatment with alpha-Klotho decreased TNF-alpha and IL-6 production, reverted NF-kappa B activation, and decreased neuronal death induced by GCM. In addition, KL incubation together with GCM + LPS completely reverts the neuronal toxicity induced by low concentration of GCM-LPS. These data suggest an anti-inflammatory and neuroprotective effect of alpha-Klotho protein in the CNS. This work demonstrated the therapeutic potential of alpha-Klotho in pathological processes which involves a neuroinflammatory component. (AU)

• FAPESP's process: 13/10787-8 - Klotho effects on insulin signaling and energetic metabolism in the central nervous system
• Grantee: Caio Henrique Yokoyama Mazucanti
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 18/14289-6 - The role of Klotho protein in Alzheimer's disease
• Grantee: Paula Fernanda Kinoshita
• Support Opportunities: Scholarships in Brazil - Post-Doctoral