Division of Histology and Embryology, International Joint Laboratory for Embryonic Development and Prenatal Medicine, Medical College, Jinan University, Guangzhou, 510632, China.
Key Laboratory of Functional and Clinical Translational Medicine, Fujian Province University, Xiamen Medical College, Fujian, 350108, China.
Sci Rep. 2024 Aug 29;14(1):20136. doi: 10.1038/s41598-024-69523-z.
Pregestational diabetes mellitus (PGDM) has an impact on fetal bone formation, but the underlying mechanism is still obscure. Although miRNAs have been extensively investigated throughout bone formation, their effects on fetal bone development caused by PGDM still need clarification. This study intends to examine the mechanism by which hyperglycemia impairs the bone formation of offspring via miR-322-5p (miR-322). In this study, miR-322 was selected by systemically screening utilizing bioinformatics and subsequent validation experiments. Using streptozotocin (STZ)-induced diabetic mice and ATDC5 cell lines, we found that miR-322 was abundantly expressed in the proliferative and hypertrophic zones of the growth plate, and its expression pattern was disturbed in the presence of hyperglycemia, suggesting that miR-322 is involved in the chondrocyte proliferation and differentiation in absence/presence of hyperglycemia. This observation was proved by manipulating miR-322 expression in ATDC5 cells by transfecting mimic and inhibitor of miR-322. Furthermore, Adamts5, Col12a1, and Cbx6 were identified as the potential target genes of miR-322, verified by the co-transfection of miR-322 inhibitor and the siRNAs, respectively. The evaluation criteria are the chondrocyte proliferation and differentiation and their relevant key gene expressions (proliferation: Sox9 and PthIh; differentiation: Runx2 and Col10a1) after manipulating the gene expressions in ATDC5 cells. This study revealed the regulative role miR-322 on chondrocyte proliferation and differentiation of growth plate by targeting Adamts5, Col12a1, and Cbx6 in hyperglycemia during pregnancy. This translational potential represents a promising avenue for advancing our understanding of bone-related complications in diabetic pregnancy and mitigating bone deficiencies in diabetic pregnant individuals, improving maternal and fetal outcomes.
孕前糖尿病（PGDM）会影响胎儿的骨骼形成，但其中的机制仍不明确。尽管 miRNA 在整个骨形成过程中已经被广泛研究，但它们对 PGDM 引起的胎儿骨骼发育的影响仍需要阐明。本研究旨在通过 miR-322-5p（miR-322）研究高血糖损害后代骨骼形成的机制。在这项研究中，我们通过系统的生物信息学筛选和后续的验证实验选择了 miR-322。利用链脲佐菌素（STZ）诱导的糖尿病小鼠和 ATDC5 细胞系，我们发现 miR-322 在生长板的增殖和肥大区大量表达，其表达模式在高血糖存在下受到干扰，表明 miR-322 参与了软骨细胞在高血糖存在/不存在的情况下的增殖和分化。这一观察结果通过在 ATDC5 细胞中转染 miR-322 的模拟物和抑制剂来操纵 miR-322 的表达得到了证实。此外，Adamts5、Col12a1 和 Cbx6 被鉴定为 miR-322 的潜在靶基因，通过分别共转染 miR-322 抑制剂和 siRNAs 得到验证。评估标准是操纵 ATDC5 细胞中的基因表达后，软骨细胞的增殖和分化及其相关关键基因的表达（增殖：Sox9 和 PthIh；分化：Runx2 和 Col10a1）。本研究揭示了 miR-322 通过靶向 Adamts5、Col12a1 和 Cbx6 在妊娠期间高血糖状态下对生长板软骨细胞增殖和分化的调节作用。这一转化潜力为深入了解糖尿病妊娠中的骨骼相关并发症以及减轻糖尿病孕妇的骨骼缺陷提供了有前途的途径，从而改善母婴结局。
