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1.海南省妇女儿童医学中心产科,海南 海口 570000
2.海南医学院第一附属医院产科,海南 海口 570000
黄赞怡,E-mail:niao298@sina.com
收稿日期:2025-02-26,
纸质出版日期:2025-08-25
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黄赞怡,符梅沙,陈惠丽等.CircPDS5B水平上调参与早产的病理生理机制研究[J].局解手术学杂志,2025,34(08):668-674.
HUANG Zan-yi,FU Mei-sha,CHEN Hui-li,et al.Up-regulation of CircPDS5B level involving in the pathophysiological mechanism of premature birth[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(08):668-674.
黄赞怡,符梅沙,陈惠丽等.CircPDS5B水平上调参与早产的病理生理机制研究[J].局解手术学杂志,2025,34(08):668-674. DOI: 10.11659/jjssx.02E025142.
HUANG Zan-yi,FU Mei-sha,CHEN Hui-li,et al.Up-regulation of CircPDS5B level involving in the pathophysiological mechanism of premature birth[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(08):668-674. DOI: 10.11659/jjssx.02E025142.
目的
2
探讨CircPDS5B水平上调参与早产的病理生理分子机制。
方法
2
收集足月儿(Group 1,孕周≥37周)、晚期早产儿(Group 2,34周≤孕周<37周)、早产儿(Group 3,32周<孕周<34周)和极早产儿(Group 4,孕周≤32周)的胎盘组织,每组5例样本。提取其总RNA并进行CircRNAs微阵列分析。qPCR法验证胎盘组织中前10位显著上调和前10位显著下调的CircRNAs的表达水平。在体外人胎盘微血管内皮细胞(hPMEC)中采用RNA免疫共沉淀法验证CircPDS5B与异质核糖核蛋白L(hnRNPL)及hnRNPL与缺氧诱导因子-1α(HIF-1α) mRNA的直接结合关系。将hPMEC分为Control组、Lenti-sh-CircPDS5B组、Lenti-sh-NT组、Lenti-OE-CircPDS5B组及Lenti-OE-Vector组,其中Control组细胞未进行转染,其余组细胞分别转染相应的慢病毒颗粒。Western blot法测定各组细胞中HIF-1α和血管内皮生长因子(VEGF)-A蛋白表达水平。
结果
2
CircRNAs微阵列分析和qPCR法验证结果显示,与Group 1相比,CircPDS5B在Group 4中上调最显著(
P
<
0.05)。RNA免疫共沉淀结果显示,CircPDS5B与hnRNPL以及hnRNPL与HIF-1α mRNA存在直接结合关系。与Lenti-sh-NT组相比,Lenti-sh-CircPDS5B组细胞中HIF-1α和VEGF-A的表达水平升高(
P
<
0.05)。与Lenti-OE-Vector组相比,Lenti-OE-CircPDS5B组细胞中HIF-1α和VEGF-A的表达水平降低(
P
<
0.05)。
结论
2
CircPDS5B水平上调通过hnRNPL结合和隔离HIF-1α mRNA抑制VEGF表达参与早产的病理生理机制。
Objective
2
To explore the pathophysiological molecular mechanism of the up-regulation of CircPDS5B level involved in premature birth.
Methods
2
The placental tissues of full-term infants (Group 1, gestational age ≥ 37 weeks), late premature infants (Group 2, 34 weeks ≤ gestational age
<
37 weeks), premature infants (Group 3, 32 weeks
<
gestational age
<
34 weeks) and very premature infants (Group 4, gestational age ≤ 32 weeks) were collected, with 5 samples in each group. Total RNA was extracted from placental tissues for CircRNAs microarray. The expression levels of the top 10 significantly up-regulated and top 10 significantly down-regulated CircRNAs in the placental tissues were verified by qPCR. The direct binding relationship between CircPDS5B and heterogenous nuclear ribonucleoprotein L(hnRNPL), as well as hnRNPL and hypoxia-induced factor-1α(HIF-1α) mRNA were verified by RNA immunocoprecipitation in human placental microvascular endothelial cells (hPMEC)
in vitro
. HPMEC were divided into Control group, Lenti-sh-CircPDS5B group, Lenti-sh-NT group, Lenti-OE-CircPDS5B group and Lenti-OE-Vector group.The cells in the Control group were not transfected, and the cells in the other groups were transfected with corresponding lentivirus particles respectively. The expression levels of HIF-1α and vascular endothelial growth factor(VEGF)-A protein of cells in each group were determined by Western blot.
Results
2
The results of CircRNAs microarray analysis and qPCR verification showed that compared with Group 1, CircPDS5B was up-regulated most significantly in Group 4 (
P
<
0.05). The RNA immunoprecipitation results showed that there was a direct binding relationship between CircPDS5B and hnRNPL, as well as hnRNPL and HIF-1α mRNA. Compared with Lenti-sh-NT group, the expression levels of HIF-1α and VEGF-A of cells in Lenti-sh-CircPDS5B group were increased (
P
<
0.05). Compared with Lenti-OE-Vector group, the expression levels of HIF-1α and VEGF-A of cells in Lenti-OE-CircPDS5B group were decreased (
P
<
0.05).
Conclusion
2
Up-regulation of CircPDS5B level is involved in the pathophysiological mechanisms of premature birth by inhibiting VEGF expression through hnRNPL binding and sequestering HIF-1α mRNA.
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