This study aimed to verify the toxic effects of prenatal caffeine exposure (PCE) on the podocyte development in male offspring, and to explore the underlying intrauterine programming mechanisms. The pregnant rats were administered with caffeine (30 to 120 mg/kg⋅d) during gestational day (GD) 9 to 20. The male fetus on GD20 and the offspring at postnatal week (PW) 6 and PW28 were sacrificed. The results indicated that PCE caused ultrastructural abnormalities on podocyte, and inhibited the expression of podocyte marker genes such as Nephrin, Wilms tumor 1 (WT1), the histone 3 lysine 9 acetylation (H3K9ac) level in the Kruppel-like factor 4 (KLF4) promoter and its expression in the male offspring from GD20 to PW28. Meanwhile, the expression of glucocorticoid receptor (GR) and histone deacetylase 7 (HDAC7) in the fetus were increased by PCE. In vitro, corticosterone increased GR and HDAC7 whereas reduced the H3K9ac level of KLF4 and KLF4/Nephrin expression. KLF4 over-expression reversed the reduction of Nephrin expression, knockdown of HDAC7 and GR antagonist RU486 partially reversed the inhibitory effects of corticosterone on H3K9ac level and KLF4 expression. In conclusion, PCE caused podocyte developmental toxicity in male offspring, which was associated with corticosterone-induced low-functional programming of KLF4 through GR/HDAC7/H3K9ac pathway.

中文翻译：

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KLF4 的 H3K9ac 水平降低介导产前咖啡因暴露诱导的雄性后代大鼠足细胞发育毒性

本研究旨在验证产前咖啡因暴露（PCE）对雄性后代足细胞发育的毒性作用，并探讨潜在的宫内编程机制。在妊娠第 9 至 20 天（GD）给怀孕的大鼠服用咖啡因（30 至 120 毫克/千克·d）。处死 GD20 的雄性胎儿和出生后第 6 周（PW）和 PW28 的后代。结果表明，PCE引起足细胞超微结构异常，抑制足细胞标志基因如Nephrin、Wilmstumour 1 (WT1)、Kruppel样因子4 (KLF4)中组蛋白3赖氨酸9乙酰化(H3K9ac)水平的表达。启动子及其在 GD20 至 PW28 雄性后代中的表达。同时，PCE增加了胎儿糖皮质激素受体（GR）和组蛋白去乙酰化酶7（HDAC7）的表达。体外，皮质酮增加了 GR 和 HDAC7，而降低了 KLF4 和 KLF4/Nephrin 表达的 H3K9ac 水平。KLF4 过表达逆转了 Nephrin 表达的降低，HDAC7 和 GR 拮抗剂 RU486 的敲低部分逆转了皮质酮对 H3K9ac 水平和 KLF4 表达的抑制作用。总之，PCE 导致雄性后代足细胞发育毒性，这与皮质酮通过 GR/HDAC7/H3K9ac 途径诱导的 KLF4 低功能编程有关。