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连翘-金银花药对调控血清外泌体miRNA对急性肺损伤的改善作用及机制研究
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| 篇名： | 连翘-金银花药对调控血清外泌体miRNA对急性肺损伤的改善作用及机制研究 |
| TITLE： | Ameliorative effect and mechanism of Forsythia suspensa-Lonicera japonica herb pair on acute lung injury via regulating serum exosomal miRNA |
| 摘要： | 目的 基于血清外泌体微RNA（miRNA），研究连翘-金银花药对对急性肺损伤（ALI）的改善作用及机制。方法将大鼠分为空白组（生理盐水）、模型组（生理盐水）、连翘-金银花药对组（2.55g/kg），每组10只。除空白组外，其余各组大鼠均通过气管滴注5mg/mL脂多糖复制ALI模型。造模后，各组大鼠灌胃相应药液/生理盐水，每天1次，连续3d。末次给药后，观察大鼠肺组织病理学形态；测定肺组织湿干重比和支气管肺泡灌洗液（BALF）中白细胞数量；检测BALF中炎症因子[肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、IL-10]水平；提取大鼠血清外泌体，利用高通量测序技术筛选外泌体中差异表达miRNA，并进行京都基因与基因组百科全书（KEGG）富集分析。基于筛选的差异表达miRNA和富集的KEGG通路，通过体外细胞实验进行机制验证。结果动物实验结果显示，经连翘-金银花药对干预后，ALI大鼠湿干重比以及BALF中白细胞数量和TNF-α、IL-1β水平均显著降低/减少（P＜0.01），IL-10水平显著升高（P＜0.01）。高通量测序实验结果显示，连翘-金银花药对可显著上调外泌体中miR-345-3p、miR-194-5p、miR-653-5p等的表达，其中差异表达miRNA的靶基因涉及的KEGG通路包括缺氧诱导因子1（HIF-1）信号通路等。细胞验证实验结果显示，过表达的miR-345-3p可显著升高细胞上清液中IL-10水平（P＜0.01），显著降低细胞上清液中TNF-α、IL-1β水平以及细胞中蛋白激酶B1、磷脂酰肌醇3-激酶、HIF-1α的mRNA及其蛋白表达水平（P＜0.01）。结论连翘-金银花药对可通过上调血清外泌体中miR-345-3p表达，抑制HIF-1信号通路活性，从而减轻炎症反应，进而发挥改善ALI的作用。 |
| ABSTRACT： | OBJECTIVE To study the ameliorative effect and mechanism of Forsythia suspensa-Lonicera japonica herb pair on acute lung injury （ALI） based on serum exosomal microRNA （miRNA）. METHODS The rats were randomly divided into a blank group （normal saline）， model group （nomal saline）， and F. suspensa-L. japonica herb pair group （2.55 g/kg）， with 10 rats in each group. Except for the blank group， the other groups were used to establish an ALI model by intratracheal dripping of 5 mg/ mL lipopolysaccharides. After modeling， each group was given relevant medicine/normal saline intragastrically， once a day， for 3 consecutive days. After the last medication， the pathological status of lung tissue was observed； lung wet-to-dry weight ratio and leukocyte counts in bronchoalveolar lavage fluid （BALF） were determined. The levels of inflammatory factors [tumor necrosis factor-α（TNF-α）， interleukin-1β （IL-1β）， IL-10] in BALF were determined. Exosomes were isolated from rat serum， and high- throughput sequencing technology was employed to screen differentially expressed miRNA within the exosomes， followed by Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis. Based on the screened differentially expressed miRNA and the enriched KEGG pathways， in vitro cellular experiments were conducted for validation. RESULTS The animal experimental results demonstrated that after intervention with the F. suspensa-L. japonica herb pair， the wet-to-dry weight ratio， the number of leukocytes in BALF， as well as the levels of TNF-α and IL-1β in BALF of ALI rats were all significantly reduced （P＜0.01）， while the level of IL-10 was significantly increased （P＜0.01）. The results of high-throughput sequencing experiments revealed that the F. suspensa-L. japonica herb pair could significantly up-regulate the expressions of miR-345-3p， miR-194-5p， miR-653-5p， and others in exosomes. Among them， the KEGG pathways involved in the target genes of differentially expressed miRNA included the hypoxia-inducible factor-1（HIF-1） signaling pathway， among others. The results of cellular E-mail：huang.haiying@126.com validation experiments showed that overexpressed miR-345-3p could significantly elevate the level of IL-10 in the cell supernatant （P＜0.01）， while significantly reducing the levels of TNF-α and IL-1β in the cell supernatant， as well as the mRNA and protein expression levels of protein kinase B1， phosphatidylinositol 3- kinase， and HIF-1α （P＜0.01）. CONCLUSIONS F. suspensa-L. japonica herb pair can alleviate inflammatory responses and thereby exert a therapeutic effect in improving ALI by up-regulating the expression of miR-345-3p in serum exosomes and inhibiting the activity of the HIF-1 signaling pathway. |
| 期刊： | 2026年第37卷第03期 |
| 作者： | 陈朝华；谢淑敏；常万顺；韩雨晴；陈艳雯；朱艳慧；曹明卓；黄海英 |
| AUTHORS： | CHEN Zhaohua， XIE Shumin，CHANG Wanshun，HAN Yuqing，CHEN Yanwen，ZHU Yanhui，CAO Mingzhuo，HUANG Haiying |
| 关键字： | 连翘-金银花药对；急性肺损伤；外泌体；miR-345-3p；HIF-1信号通路；高通量测序技术 |
| KEYWORDS： | Forsythia suspensa-Lonicera japonica herb pair； acute lung injury； exosomes； miR-345-3p； HIF-1 signaling |
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