Zhang Meng, Zhang Mingming
Pediatric Outpatient Infusion Room, Xuzhou Central Hospital, Xuzhou, 221009, Jiangsu, China.
Department of Pediatrics, Xuzhou Central Hospital, No. 199 Jiefang South Road, Xuzhou, 221009, Jiangsu, China.
BMC Pediatr. 2025 May 28;25(1):428. doi: 10.1186/s12887-025-05766-7.
This study sought to investigate the role and molecular mechanisms of nucleotide-binding oligomerization domain (NOD)-like receptor family caspase activation and recruitment domain (CARD)-containing 3 (NLRC3) in the inflammatory responses of neonates, thereby developing new clinical insights into the occurrence and prevention of neonatal infections.
Peripheral blood samples were collected from full-term infants (n = 49) and preterm infants (n = 41) without any signs of intrauterine infection, as well as from healthy non-pregnant adults (n = 45). A real-time polymerase chain reaction was used to assess the expression levels of NLRC3 and NOD-containing protein 1 (NOD1) in the isolated mononuclear cells. Whole blood from the adults, full-term infants, and preterm infants was stimulated for four hours with a mixture of herpes simplex virus type 60 DNA (HSV-60 DNA) and lipopolysaccharides (LPS) or LPS alone or blank medium. An enzyme-linked immunosorbent assay was employed to measure the tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and interleukin 1 beta (IL-1β) levels in the supernatant.
The gene expression levels of NLRC3 were significantly lower in the full-term and preterm infants than in the adults, with the preterm infants showing notably lower levels when compared with the full-term infants. A positive correlation was found between the NLRC3 and NOD1 expression levels in the neonates (both full-term and preterm), indicating lower NLRC3 expression to be associated with lower NOD1 expression. After LPS stimulation, the production of TNF-α, IL-6, and IL-1β in the whole blood of the preterm and full-term infants was significantly lower than in that of the adults. Moreover, stimulation with a combination of LPS and HSV-60 DNA resulted in similar TNF-α, IL-6, and IL-1β production across the blood samples from preterm infants, full-term infants, and adults. When compared with LPS stimulation alone, the LPS and HSV-60 DNA mixture significantly reduced the release of TNF-α, IL-6, and IL-1β in the adults. In the neonates, however, only the release of TNF-α was significantly reduced, as no notable difference was observed in the IL-6 and IL-1β levels.
The reduced expression and functional impairment of NOD-like receptors, such as NLRC3 and NOD1, in neonates, may contribute to their heightened susceptibility to severe infections. This finding indicates new avenues for the prevention and treatment of neonatal infections.
本研究旨在探讨核苷酸结合寡聚化结构域（NOD）样受体家族含半胱天冬酶激活和募集结构域（CARD）3（NLRC3）在新生儿炎症反应中的作用及分子机制，从而为新生儿感染的发生和预防提供新的临床见解。
采集足月婴儿（n = 49）、早产婴儿（n = 41）外周血样本，这些婴儿无任何宫内感染迹象，同时采集健康未孕成人（n = 45）外周血样本。采用实时聚合酶链反应评估分离出的单核细胞中NLRC3和含NOD蛋白1（NOD1）的表达水平。用单纯疱疹病毒60型DNA（HSV - 60 DNA）和脂多糖（LPS）混合物、单独的LPS或空白培养基刺激成人、足月婴儿和早产婴儿的全血4小时。采用酶联免疫吸附测定法测量上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）和白细胞介素1β（IL-1β）水平。
足月和早产婴儿中NLRC3的基因表达水平显著低于成人，与足月婴儿相比，早产婴儿的NLRC3水平明显更低。在新生儿（足月和早产）中，NLRC3和NOD1表达水平呈正相关，表明NLRC3表达降低与NOD1表达降低相关。LPS刺激后，早产和足月婴儿全血中TNF-α、IL-6和IL-1β的产生显著低于成人。此外，LPS和HSV - 60 DNA联合刺激导致早产婴儿、足月婴儿和成人血样中TNF-α、IL-6和IL-1β的产生相似。与单独LPS刺激相比，LPS和HSV - 60 DNA混合物显著降低了成人中TNF-α、IL-6和IL-1β的释放。然而，在新生儿中，仅TNF-α的释放显著降低，IL-6和IL-1β水平未观察到明显差异。
新生儿中NLRC3和NOD1等NOD样受体的表达降低和功能受损可能导致其对严重感染的易感性增加。这一发现为新生儿感染的预防和治疗指明了新途径。
