Polyhexamethylene guanidine phosphate (PHMG-p) was used as a humidifier disinfectant in Korea. PHMG induced severe pulmonary fibrosis in Koreans. The objective of this study was to elucidate mechanism of pulmonary toxicity caused by PHMG-p in rats using multi-omics analysis. Wistar rats were intratracheally instilled with PHMG-p by single (1.5 mg/kg) administration or 4-week (0.1 mg/kg, 2 times/week) repeated administration. Histopathologic examination was performed with hematoxylin and eosin staining. Alveolar macrophage aggregation and granulomatous inflammation were observed in rats treated with single dose of PHMG-p. Pulmonary fibrosis, chronic inflammation, bronchiol-alveolar fibrosis, and metaplasia of squamous cell were observed in repeated dose group. Next generation sequencing (NGS) was performed for transcriptome profiling after mRNA isolation from bronchiol-alveoli. Bronchiol-alveoli proteomic profiling was performed using an Orbitrap Q-exactive mass spectrometer. Serum and urinary metabolites were determined using 1H-NMR. Among 418 differentially expressed genes (DEGs) and 67 differentially expressed proteins (DEPs), changes of 16 mRNA levels were significantly correlated with changes of their protein levels in both single and repeated dose groups. Remarkable biological processes represented by both DEGs and DEPs were defense response, inflammatory response, response to stress, and immune response. Arginase 1 (Arg1) and lipocalin 2 (Lcn2) were identified to be major regulators for PHMG-p-induced pulmonary toxicity based on merged analysis using DEGs and DEPs. In metabolomics study, 52 metabolites (VIP > 0.5) were determined in serum and urine of single and repeated-dose groups. Glutamate and choline were selected as major metabolites. They were found to be major factors affecting inflammatory response in association with DEGs and DEPs. Arg1 and Lcn2 were suggested to be major gene and protein related to pulmonary damage by PHMG-p while serum or urinary glutamate and choline were endogenous metabolites related to pulmonary damage by PHMG-p.

中文翻译：

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转录组学，蛋白质组学和代谢组学的整合确定了大鼠加湿消毒剂聚六亚甲基磷酸胍（PHMG-p）对肺损伤的生物标记。

在韩国，聚六亚甲基磷酸胍（PHMG-p）被用作加湿器消毒剂。PHMG导致韩国人严重肺纤维化。这项研究的目的是使用多组学分析阐明由PHMG-p引起的大鼠肺毒性的机制。通过单次（1.5 mg / kg）给药或4周（0.1 mg / kg，2次/周）重复给药向Wistar大鼠气管内滴注PHMG-p。用苏木精和曙红染色进行组织病理学检查。在单剂量PHMG-p治疗的大鼠中观察到肺泡巨噬细胞聚集和肉芽肿性炎症。重复给药组观察到肺纤维化，慢性炎症，细支气管肺泡纤维化和鳞状上皮化生。从细支气管肺泡分离mRNA后，进行下一代测序（NGS）进行转录组分析。使用Orbitrap Q活性质谱仪进行细支气管肺泡蛋白组学分析。使用1 H-NMR测定血清和尿液代谢产物。在418个差异表达基因（DEGs）和67个差异表达蛋白（DEPs）中，单剂量和重复剂量组中16种mRNA水平的变化与其蛋白质水平的变化均显着相关。DEGs和DEPs代表的显着生物学过程是防御反应，炎症反应，应激反应和免疫反应。基于DEG和DEP的合并分析，精氨酸酶1（Arg1）和脂蛋白2（Lcn2）被确定为PHMG-p诱导的肺毒性的主要调节剂。在代谢组学研究中 在单次和重复剂量组的血清和尿液中测定了52种代谢产物（VIP> 0.5）。选择谷氨酸和胆碱作为主要代谢产物。发现它们是与DEG和DEP相关的影响炎症反应的主要因素。Arg1和Lcn2被认为是PHMG-p与肺损伤有关的主要基因和蛋白，而血清或尿谷氨酸和胆碱是PHMG-p与肺损伤有关的内源性代谢产物。