目的 基于转录组学研究小檗碱抗脂多糖（LPS）致肺损伤的作用及机制，并应用实时荧光定量PCR（qRT-PCR）验证筛选出的基因的表达。方法 随机将27只小鼠分为3组（每组9只）：对照组、模型（LPS，3.5 mg·kg_-1）组、小檗碱（10 mg·kg_-1）组，模型组和小檗碱组鼻腔使用移液枪吸入0.9%氯化钠溶液配制的LPS溶液，对照组吸入等量0.9%氯化钠溶液，轻晃小鼠使溶液均匀分布，每天给药1次，连续7 d。处死小鼠收集肺组织，苏木精-伊红（HE）染色观察肺组织病理变化；进行转录组学检测，通过样本间基因表达量相关性分析判断聚类情况，基于差异倍数和q值来精确筛选差异表达的基因，差异基因进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）富集分析，利用STRING数据库对差异mRNA进行网络互作分析； qRT-PCR检测右肺中下叶肌球蛋白结合蛋白H（Mybph）、肌球蛋白轻链激酶2（Mylk2）、趋化因子（CC基序）受体3（Ccr3） mRNA表达水平。结果 与模型组相比，小檗碱组肺组织病理损伤明显改善，单位面积肺泡周长、单位面积肺泡数量、单位面积肺泡面积、肺泡面积百分比显著上升（P＜0.001），纤维化评分和炎症评分显著下降（P＜0.001）。转录组学相关性分析表明小檗碱组与对照组基因表达相似，与模型组差异较大；对照组vs模型组和模型组vs小檗碱组2个比较组共同调控274个基因，其中，相较于对照组，模型组上调了227个基因、下调了47个基因，小檗碱组能够将这些基因表达恢复到接近对照组的水平； GO富集分析表明，对照组vs模型组和模型组vs小檗碱组共同调控的过程有炎症反应（inflammatory response）、G蛋白偶联受体信号通路（G protein-coupled receptor signaling pathway）、免疫反应（immune response）、细胞黏附（cell adhesion）、磷酸化（phosphorylation）、细胞质（cytoplasm）、质膜（plasma membrane）、蛋白质结合（protein binding）、核苷酸结合（nucleotide binding）、ATP结合（ATP binding）等； KEGG分析结果聚焦于黏着斑（Focal adhesion）、肌动蛋白细胞骨架的调控（Regulation of actin cytoskeleton）、细胞因子-细胞因子受体相互作用（Cytokine-cytokine receptor interaction）、钙信号通路（Calcium signaling pathway）、趋化因子信号通路（Chemokine signaling pathway）等； GO、KEGG富集分析差异mRNA互作网络中的共同基因为Mybph、Mylk2、Ccr3。qRT-PCR验证结果表明，Mybph、Mylk2表达在3个处理组中差异无统计学意义。与对照组相比，模型组Ccr3 mRNA的表达水平显著增加（P＜0.001）；与模型组相比，小檗碱能显著降低Ccr3的表达（P＜0.001）。结论 小檗碱可能通过抑制Ccr3的表达，减轻LPS所致肺部损伤。

Objective The effect and mechanism of berberine on lipopolysaccharide (LPS)-induced lung injury were investigated based on transcriptomics, and the selected genes were verified by real-time fluorescence quantitative PCR (qRT-PCR). Methods A total of 27 mice were randomly divided into three groups (9 mice in each group): control group, model (LPS, 3.5 mg·kg^-1) group, and berberine (10 mg·kg^-1) group. The model group and the berberine group were intranasally administered LPS solution prepared with 0.9% sodium chloride solution using a pipette, while the control group was administered the same volume of 0.9% sodium chloride solution. The mice were gently shaken to ensure uniform distribution of the solution. The administration was performed once a day for seven consecutive days. The mice were sacrificed and lung tissues were collected. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in lung tissues. Transcriptomics was conducted, and the clustering situation was determined by analyzing the correlation of gene expression levels between samples. Differentially expressed genes were precisely screened based on fold change and q value. GO enrichment analysis and KEGG enrichment analysis were performed on the differentially expressed genes. The STRING database was used to analyze the network interaction of differentially expressed mRNAs. The mRNA expression levels of myosin binding protein H (Mybph), myosin light chain kinase 2 (Mylk2), and chemokine (CC motif) receptor 3 (Ccr3) in the lower lobe of the right lung were detected by qRT-PCR. Results Compared with the model group, the pathological damage of lung tissue in the berberine group was significantly improved. The alveolar perimeter per unit area, the number of alveoli per unit area, the alveolar area per unit area, and the percentage of alveolar area significantly increased (P < 0.001), while the fibrosis score and inflammation score significantly decreased (P < 0.001). Transcriptomics correlation analysis indicated that the gene expression in the berberine group was similar to that in the control group and significantly different from that in the model group. The two comparison groups, control group vs. model group and model group vs. berberine group, jointly regulated 274 genes. Compared with the control group, the model group upregulated 227 genes and downregulated 47 genes, and berberine could restore the expression of these genes to a level close to that of the control group. GO enrichment analysis showed that the jointly regulated processes by the control group vs model group and model group vs berberine group included inflammatory response, G protein-coupled receptor signaling pathway, immune response, cell adhesion, phosphorylation, cytoplasm, plasma membrane, protein binding, nucleotide binding, and ATP binding, etc. KEGG analysis results focused on focal adhesion, regulation of actin cytoskeleton, cytokine-cytokine receptor interaction, calcium signaling pathway, and chemokine signaling pathway, etc. The common genes in the GO and KEGG enrichment analysis of the interaction network of differentially expressed mRNAs were Mybph, Mylk2, and Ccr3. The results of qRT-PCR validation indicated that there was no statistically significant difference in the expression of Mybph and Mylk2 among the three treatment groups. Compared with the control group, the expression level of Ccr3 mRNA in the model group was significantly increased (P < 0.001); Compared with the model group, berberine could significantly reduce the expression of Ccr3 (P < 0.001). Conclusion Berberine may alleviate LPSinduced lung injury by inhibiting the expression of Ccr3.

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内蒙古自然科学基金资助项目（2024MS08048）