Rosmarinic acid relieves LPS-induced sickness and depressive-like behaviors in mice by activating the BDNF/Nrf2 signaling and autophagy pathway

Abstract

Neuroinflammation is one of the main causes of sickness and depressive-like behavior. Rosmarinic acid (RA) has been shown to have a significant anti-neuroinflammatory effect. However, the protective effects and the underlying mechanism of RA on sickness and depressive-like behavior under conditions of neuroinflammation are still unclear. In the present study, we investigated the effects and the underlying mechanism of RA on lipopolysaccharide (LPS)-treated mice with sickness behavior. The behavioral effects of LPS treatment and RA administration were assessed using behavioral tests including a sucrose preference test and an open field test. The neuroprotective effects of RA in conditions of neuroinflammatory injury were determined by HE staining, Nissl staining, and immunofluorescent staining. Moreover, its underlying mechanism was analyzed by using real-time PCR analysis, western blot, and immunofluorescent analysis. The results indicated that RA dramatically mitigated sickness behaviors and histologic brain damage in mice exposed to LPS. In addition, RA administration markedly promoted the expression of brain-derived neurotrophic factor (BDNF)/erythroid 2-related factor 2 (Nrf2), the key regulatory proteins for Nrf2 activation (p21 and p62), the downstream antioxidant enzymes (HO-1, NQO1, GCLC), the autophagy-related proteins (LC3II and Beclin1), and mitochondrial respiratory enzyme genes (ME1, IDH1, 6-PGDH), while reducing the expression of pro-inflammatory genes (CD44, iNOS, TNFα, IL-1β). Moreover, the double-label immunofluorescent analysis revealed that RA increased the fluorescence intensity of LC3 mostly co-localized with neurons and co-expressed with Nrf2. Taken together, our research found that RA could effectively alleviate sickness behaviors and nerve injury caused by neuroinflammation, and its protective effects were mediated by the Nrf2 signaling pathway, which reduced cellular oxidative stress, inflammation, mitochondrial respiratory function damage, and autophagy imbalance. Therefore, RA has the potential to prevent or treat sickness and depressive-like behaviors under conditions of neuroinflammation.

Keywords: Autophagy pathway; BDNF/Nrf2 signaling; Depression; Lipopolysaccharide; Mitochondrial respiration; Oxidative stress; Rosmarinic acid.