脂多糖诱导早产大鼠宫颈重塑障碍的转录组学分析

Transcriptomic analysis of lipopolysaccharide-induced cervical remodeling dysfunction in a rat model of preterm birth

摘要:

目的 通过转录组学测序分析脂多糖(lipopolysaccharide, LPS)诱导的早产大鼠模型中宫颈组织基因表达变化及潜在作用机制。方法 对腹腔注射LPS和PBS孕鼠宫颈组织进行转录组测序分析,对差异表达基因进行GO分析和KEGG通路富集分析,寻找与宫颈重塑、炎症、细胞外基质、缓激肽相关的关键基因,并通过qRT-PCR进行验证。结果 LPS组孕鼠均发生早产(100% vs 0%,P=0.000 5)、妊娠期显著缩短[(18.08±0.38) d vs (21.92±0.58) d,P<0.000 1]。转录组分析显示,LPS组宫颈组织中有892个基因上调、725个基因下调。GO分析显示差异基因富集于免疫反应、细胞外基质和趋化因子活性等通路;KEGG分析表明炎症反应及缓激肽系统可能破坏细胞外基质的稳定,诱发宫颈机能不全相关疾病,导致早产发生。对宫颈重塑关键基因进行qRT-PCR验证,显示IL-6、TNF-α、COL3A1、LOX、MMP9、PTGS2、KNG1、MME在LPS组中均上调,与转录组学检测结果一致。结论 LPS通过激活炎症反应和缓激肽系统,协同调控PTGS2/PGE2通路及宫颈重塑相关基因,导致细胞外基质紊乱、宫颈机能不全,最终诱发早产。

Abstract:

Objective To analyze the gene expression changes and the potential mechanisms in cervical tissue of a lipopolysaccharide(LPS)-induced rat model of preterm birth using transcriptomic sequencing. Methods Transcriptome sequencing was performed on cervical tissues from pregnant rats administered with intraperitoneal LPS or phosphate buffered saline(PBS) injections. Differentially expressed genes(DEGs) were analyzed through gene ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analyses to identify key genes associated with cervical remodeling, inflammation, extracellular matrix, and the bradykinin system. The genes were validated with quantitative reverse transcription polymerase chain reaction(qRT-PCR). Results All LPS-treated rats experienced preterm birth(100% vs 0%, P=0.0005) with significantly shortened gestation periods [(18.08±0.38) days vs (21.92±0.58) days, P<0.0001]. Transcriptomic analysis revealed 892 upregulated and 725 downregulated genes in the LPS group. The GO analysis showed that DEGs were enriched in immune response, extracellular matrix and chemokine activity pathways. The KEGG analysis indicated that inflammatory responses and the bradykinin system may disrupt extracellular matrix stability, leading to disorders associated cervical insufficiency and preterm birth. The QRT-PCR validation of key cervical remodeling genes confirmed upregulation of interleukin(IL)-6, tumor necrosis factor (TNF)-α, collagen type 3 α1(COL3A1), lysyl oxidase (LOX), matrix metalloproteinase 9(MMP9), prostaglandin endoperoxide synthase 2(PTGS2), kininogen 1(KNG1), and membrane metalloendopeptidase(MME) in the LPS group, consistent with transcriptomic findings. Conclusion LPS triggers preterm birth by activating inflammatory responses and the bradykinin system, synergistically regulating the PTGS2/PGE2 pathway and cervical remodelingrelated genes, ultimately causing extracellular matrix dysfunction and cervical insufficiency.