Department of Joint Surgery and Sports Medicine, Zhongnan Hospital of Wuhan University, Wuhan, China.
Department of Orthopedics Surgery, Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, China.
FASEB J. 2021 Feb;35(2):e21370. doi: 10.1096/fj.202002230R.
Caffeine has developmental toxicity. Prenatal caffeine exposure (PCE) caused intrauterine growth retardation (IUGR) and multiple organ dysplasia. This study intended to explore the effect and mechanism of PCE on long bone development in female fetal rats. In vivo, the PCE group pregnant rats were given different concentrations of caffeine during the gestational Day 9-20. The mRNA expression of osteogenesis-related genes were significantly reduced in PCE group. In the PCE group (120 mg/kg·d), the length and primary center of fetal femur were shorter, and accompanied by H-type blood vessel abundance reducing. Meanwhile, connective tissue growth factor (CTGF) expression decreased in the growth plate of the PCE group (120 mg/kg·d). In contrast, the miR375 expression increased. In vitro, caffeine decreased CTGF and increased miR375 expression in fetal growth plate chondrocytes. After co-culture with caffeine-treated chondrocytes, the tube formation ability for the H-type endothelial cells was decreased. Furthermore, CTGF overexpression or miR375 inhibitor reversed caffeine-induced reduction of tube formation ability, and miR375 inhibitor reversed caffeine-induced CTGF expression inhibition. In summary, PCE decreased the expression of CTGF by miR375, ultimately resulting in H-type blood vessel-related long bone dysplasia.
咖啡因具有发育毒性。产前咖啡因暴露（PCE）可导致宫内生长迟缓（IUGR）和多器官发育不良。本研究旨在探讨 PCE 对雌性胎鼠长骨发育的影响及其机制。在体内，PCE 组孕鼠在妊娠第 9-20 天给予不同浓度的咖啡因。PCE 组的成骨相关基因的 mRNA 表达明显降低。在 PCE 组（120mg/kg·d）中，胎鼠股骨的长度和初级中心变短，伴随着 H 型血管丰度减少。同时，PCE 组（120mg/kg·d）生长板中的结缔组织生长因子（CTGF）表达减少。相比之下，miR375 的表达增加。在体外，咖啡因降低了胎生长板软骨细胞中的 CTGF 并增加了 miR375 的表达。与用咖啡因处理的软骨细胞共培养后，H 型内皮细胞的管形成能力降低。此外，CTGF 过表达或 miR375 抑制剂逆转了咖啡因诱导的管形成能力降低，而 miR375 抑制剂逆转了咖啡因诱导的 CTGF 表达抑制。综上所述，PCE 通过 miR375 降低 CTGF 的表达，最终导致与 H 型血管相关的长骨发育不良。
