Xiaoyao San attenuates hepatic steatosis through estrogen receptor α pathway in ovariectomized ApoE-/- mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyao San (XYS) is a famous prescription in traditional Chinese medicine, which is used in the treatment of "liver depression and spleen deficiency" syndrome. It is often used clinically to treat chronic hepatitis, liver cirrhosis, various symptoms of postmenopausal women, especially mental disorders and digestive system diseases. However, the effect of XYS on hepatic steatosis in postmenopausal women remains unclear. In this research, we investigated the effects of XYS on hepatic steatosis in ovariectomized (OVX) apolipoprotein E knockout (ApoE-/-) mice, as well as the molecular mechanisms in vitro and in vivo. MATERIALS AND METHODS: Fifty female ApoE-/- mice were divided into 5 groups: control group (Sham), model group (OVX), OVX + ß-estradiol (E2, 0.4 mg/kg) group, OVX + XYS (13.0 g/kg) group, and OVX + XYS (6.5 g/kg) group. The control group received a standard diet, while the other groups received a high-fat diet (HFD). The hepatic pathologies of the mice were examined with Oil red O staining and HE staining after 12 week treatment. Blood and liver variables were determined enzymatically. Transmission electron microscopy was used to examine the ultrastructure of hepatocytes. The expression of estrogen receptor α (ERα) and lipid metabolism genes was analyzed by real-time PCR and/or Western blot. In in vitro studies, we investigated the effect of XYS-medicated serum on the expression and activity of ERα in L02 cells by immunofluorescence and luciferase reporter assays, and examined the protection of XYS-medicated serum against free fatty acid (FFA)-induced steatosis of L02 cells. Intracellular lipid accumulation were measured by Oil red O staining and Nile red staining assay. Finally, the influence of ICI 182,780, a specific antagonist of ERα, on the protective effect of XYS-medicated serum on FFA-induced steatosis of L02 cells was investigated. RESULTS: Treatment of Ovx/ApoE-/- mice with XYS significantly decreased HFD-induced increases in hepatic steatosis and triglyceride (TG) content, accompanied by inhibition of liver X receptor α (LXRα), sterol regulatory element binding protein (SREBP)-1c and its target lipogenic genes transcription. Similarly, XYS-medicated serum reduced the size and number of lipid droplets and the cellular TG content in FFA-induced L02 cells. In addition, XYS significantly increased the ERα expression in hepatocytes in vivo and in vitro and enhanced the transcriptional activity of ERα promoter in L02 cells. And these effects could be partly reversed by the antiestrogen ICI 182,780. CONCLUSIONS: These findings suggest that XYS has an estrogen-like effect and inhibits steatosis in postmenopausal animal models by reducing the expression of genes related to TG synthesis through ERα pathway.

Influenza vaccination in early Alzheimer's disease rescues amyloidosis and ameliorates cognitive deficits in APP/PS1 mice by inhibiting regulatory T cells.

BACKGROUND: Alzheimer's disease (AD) is a neurodegenerative disorder strongly correlated with a dysfunctional immune system. Our previous results demonstrated that inactivated influenza vaccine (IIV) facilitates hippocampal neurogenesis and blocks lipopolysaccharide (LPS)-induced cognitive impairment. However, whether IIV improves cognitive deficits in an AD mouse model remains unclear. In addition, early interventions in AD have been encouraged in recent years. Here, we investigated whether IIV immunization at the preclinical stage of AD alters the brain pathology and cognitive deficits in an APP/ PS1 mouse model. METHODS: We assessed spatial learning and memory using Morris water maze (MWM). The brain ß-amyloid (Aß) plaque burden and activated microglia were investigated by immunohistochemistry. Furthermore, flow cytometry was utilized to analyze the proportions of Treg cells in the spleen. A cytokine antibody array was performed to measure the alteration of cytokines in the brain and peripheral immune system. RESULTS: Five IIV immunizations activated microglia, reduced the Aß burden and improved the cognitive impairment. Simultaneously, the IIV-induced immune response broke peripheral immunosuppression by reducing Foxp3+ regulatory T cell (Treg) activities, whereas the restoration of Treg level in the periphery using all-trans retinoic acid (ATRA) blunted the protective effects of IIV on Aß burden and cognitive functions. Interestingly, IIV immunization might increase proinflammatory and anti-inflammatory cytokine expression in the brain of APP/PS1 mice, enhanced microglial activation, and enhanced the clustering and phagocytosis of Aß, thereby creating new homeostasis in the disordered immune microenvironment. CONCLUSIONS: Altogether, our results suggest that early multiple IIV immunizations exert a beneficial immunomodulatory effect in APP/PS1 mice by breaking Treg-mediated systemic immune tolerance, maintaining the activation of microglia and removing of Aß plaques, eventually improving cognitive deficits.

Prenatal influenza vaccination rescues impairments of social behavior and lamination in a mouse model of autism.

BACKGROUND: Prenatal infection is a substantial risk factor for neurodevelopmental disorders such as autism in offspring. We have previously reported that influenza vaccination (VAC) during early pregnancy contributes to neurogenesis and behavioral function in offspring. RESULTS: Here, we probe the efficacy of VAC pretreatment on autism-like behaviors in a lipopolysaccharide (LPS)-induced maternal immune activation (MIA) mouse model. We show that VAC improves abnormal fetal brain cytoarchitecture and lamination, an effect associated with promotion of intermediate progenitor cell differentiation in MIA fetal brain. These beneficial effects are sufficient to prevent social deficits in adult MIA offspring. Furthermore, whole-genome analysis suggests a strong interaction between Ikzf1 (IKAROS family zinc-finger 1) and neuronal differentiation. Intriguingly, VAC rescues excessive microglial Ikzf1 expression and attenuates microglial inflammatory responses in the MIA fetal brain. CONCLUSIONS: Our study implies that a preprocessed influenza vaccination prevents maternal bacterial infection from causing neocortical lamination impairments and autism-related behaviors in offspring.