Rabbit Anti-Human Histone H3 (Acetyl-Lys14) monoclonal antibody, clone SN240

Our previous studies showed that prenatal ethanol exposure (PEE) elevated blood total cholesterol (TCH) level in adult offspring rats. This study was aimed at elucidating the intrauterine programming mechanism of hypercholesterolemia in adult rats induced by PEE. Pregnant Wistar rats were intragastrically administered ethanol (4 mg/kg.d) from gestational day (GD) 9 to 20. The offspring rats were euthanized at GD20 and postnatal week 24. Results showed that PEE decreased serum TCH and HDL-C levels (female and male) as well as LDL-C level (female only) in fetal rats but increased serum TCH level and the TCH/HDL-C and LDL-C/HDL-C ratios in adult rats. Furthermore, PEE elevated serum corticosterone levels but inhibited hepatic insulin-like growth factor 1 (IGF1) signaling pathway, cholesterol synthesis and output in fetal rats. The conversed changes were observed in adult rats. Moreover, histone acetylation (H3K9ac and H3K14ac) and expression of hepatic reverse cholesterol transport (RCT) related genes, scavenger receptor BI and low-density lipoprotein receptor were decreased before and after birth by PEE. In HepG2 cells, cortisol negatively regulated the IGF1 signaling pathway and cholesterol metabolic genes, but this inhibition of the cholesterol metabolic genes could be reversed by glucocorticoid receptor antagonist RU486, whereas exogenous IGF1 treatment only reversed the downregulation of RCT genes by cortisol. We confirmed a "two programming" mechanism for PEE-induced hypercholesterolemia in adult rats. The "first programming" was a glucocorticoid (GC)-induced persistent reduction of RCT genes by epigenetic modifications, and the "second programming" was the negative regulation of cholesterol synthesis and output by the GC-IGF1 axis.

Prenatal ethanol exposure (PEE) could affect offspring's testicular development. This study aimed to illuminate its intrauterine origin and the programming mechanism caused by PEE. Pregnant Wistar rats were given ethanol (4g/kg.d) by gavage administration during gestational days (GD) 9-20. Serum samples and testes of male offspring rats were collected on GD20, postnatal week (PW) 6, and PW12. We found that PEE induced testicular morphological abnormality, low serum testosterone levels, expressive suppression of 3-hydroxysteroid dehydrogenase (3-HSD), and low acetylation levels of histone 3 lysine 14 (H3K14ac) of 3-HSD before and after birth. In utero, when fetal rats were overexposed to corticosterone by PEE, the expression levels of testicular glucocorticoid receptor (GR) and histone deacetylase 2 (HDAC2) were increased, while that of steroidogenic factor 1 (SF1) was decreased. In vitro, corticosterone (rather than ethanol) at 500 to 2,000nM concentration decreased testosterone production and 3-HSD expression in a concentration-dependent manner. Moreover, corticosterone downregulated SF1 and upregulated HDAC2 via activating GR, accompanied by a low H3K14ac level of 3-HSD; SF1 overexpression could reverse the increased HDAC2 expression, and knockdown of HDAC2 could partially reverse the inhibitory effects of corticosterone on H3K14ac level and 3-HSD expression but not on SF1 expression. Taken together, PEE caused testicular dysplasia in male offspring rats, which was associated with corticosterone-induced low-functional programming of 3-HSD through the GR/SF1/HDAC2/H3K14ac pathway. This study provides new academic perspectives to illuminate the theory of Developmental Origins of Health and Disease.'