Berberine Ameliorates Prenatal Dihydrotestosterone Exposure-Induced Autism-Like Behavior by Suppression of Androgen Receptor.

Dongfang Xiang,Xuefang Liang,Chongxia Wei,Jianping Lu,Xiaofan Cai,Yongxia Wang,Mingtao Xu,Min Liu,Yujie Liang,Zichen Wang,Min Wang,Paul Yao,Ling Li

doi:10.3389/fncel.2020.00087

Abstract

Many epidemiology studies have shown that maternal polycystic ovary syndrome (PCOS) results in a greater risk of autism spectrum disorders (ASD) development, although the detailed mechanism remains unclear. In this study, we aimed to investigate the potential mechanism and provide a possible treatment for PCOS-mediated ASD through three experiments: Experiment 1: real-time PCR and western blots were employed to measure gene expression in human neurons, and the luciferase reporter assay and chromatin immunoprecipitation (ChIP) was used to map the responsive elements on related gene promoters. Experiment 2: pregnant dams were prenatally exposed to dihydrotestosterone (DHT), androgen receptor (AR) knockdown (shAR) in the amygdala, or berberine (BBR), and the subsequent male offspring were used for autism-like behavior (ALB) assay followed by biomedical analysis, including gene expression, oxidative stress, and mitochondrial function. Experiment 3: the male offspring from prenatal DHT exposed dams were postnatally treated by either shAR or BBR, and the offspring were used for ALB assay followed by biomedical analysis. Our findings showed that DHT treatment suppresses the expression of estrogen receptor β (ERβ) and superoxide dismutase 2 (SOD2) through AR-mediated hypermethylation on the ERβ promoter, and BBR treatment suppresses AR expression through hypermethylation on the AR promoter. Prenatal DHT treatment induces ERβ suppression, oxidative stress and mitochondria dysfunction in the amygdala with subsequent ALB behavior in male offspring, and AR knockdown partly diminishes this effect. Furthermore, both prenatal and postnatal treatment of BBR partly restores prenatal DHT exposure-mediated ALB. In conclusion, DHT suppresses ERβ expression through the AR signaling pathway by hypermethylation on the ERβ promoter, and BBR restores this effect through AR suppression. Prenatal DHT exposure induces ALB in offspring through AR-mediated ERβ suppression, and both prenatal and postnatal treatment of BBR ameliorates this effect. We conclude that BBR ameliorates prenatal DHT exposure-induced ALB through AR suppression, this study may help elucidate the potential mechanism and identify a potential treatment through using BBR for PCOS-mediated ASD.

Highlights

Autism spectrum disorders (ASD) are a group of neurodevelopmental disorders characterized by impaired social communication and interaction, together with restrictive and repetitive behaviors (Rossignol and Frye, 2012; Bralten et al, 2018; Hoirisch-Clapauch and Nardi, 2019)
The human ACS-5003 neurons were treated by either control (CTL), DHT (10 nM), or DHT with androgen receptor (AR) knockdown by lentivirus (DHT/shAR), or CTL with AR overexpression by lentivirus (CTL/↑AR), CTL with 10 μM BBR (CTL/BBR), or DHT with BBR (DHT/BBR) for 24 h, cells were harvested for gene expression analysis
Our results showed that DHT treatment decreased expression of superoxide dismutase 2 (SOD2) and estrogen receptor β (ERβ) to 66% and 51%, respectively compared to the CTL group, while it did not have any effect on AR expression

Summary

Introduction

Autism spectrum disorders (ASD) are a group of neurodevelopmental disorders characterized by impaired social communication and interaction, together with restrictive and repetitive behaviors (Rossignol and Frye, 2012; Bralten et al, 2018; Hoirisch-Clapauch and Nardi, 2019). Our recent studies have shown that suppression of estrogen receptor β (ERβ) and its target gene is associated with ASD development, while the expression of estrogen receptor α (ERα) has little correlation (Zou et al, 2017; Li et al, 2018; Xie et al, 2018), leading us to suppose that prenatal androgen exposure may induce ASD development through ERβ suppression. Many epidemiology studies have shown that maternal PCOS is associated with a greater risk of ASD (Kosidou et al, 2016; Cherskov et al, 2018; Katsigianni et al, 2019); prenatal androgen exposure may affect the neurodevelopment of offspring, resulting in ASD and attention-deficit/hyperactivity disorder (ADHD; Baron-Cohen et al, 2011; Berni et al, 2018; Cesta et al, 2020), while the detailed mechanism for PCOS-mediated ASD remains unclear. We chose the DHT-induced rat as the PCOS research model

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