目的: 探讨蒙药扫日劳-4(saorilao-4, SRL-4)对参与肺纤维化(pulmonary fibrosis, PF)大鼠肺组织基因调控网络微小RNAs(microRNA, miRNA)以及核心基因的影响。方法: 将大鼠随机分为4组,包括空白对照组(CON)、模型组(MOD)、阳性药对照组和SRL-4组。除CON外,其他组大鼠都通过气管内缓慢注射博来霉素来建立PF模型。取大鼠肺组织,提取总RNA进行转录组测序,使用差异分析软件edge R筛选各组差异表达的miRNA(differentially expressed miRNA, DEM),使用miRanda对DEM的靶基因(differentially expressed gene, DEG)进行预测,利用GO和KEGG对DEG进行生物功能富集分析,使用Cytoscape构建靶基因调控网络,筛选核心基因。结果: MOD与CON相比,筛选出16个DEM,SRL-4和MOD相比,筛选出10个DEM,调控的靶基因有63 052个。GO分析显示SRL-4和MOD的DEG富集在52个GO条目;KEGG分析显示DEG富集于182个信号通路,其中与嘌呤代谢通路相关的基因数最多。通过构建基因调控网络筛选出6个核心基因,即Spata25、Sultan1a1、Mpv17i、Cryba4、Jakmip3和Fkbp5。结论: 通过构建SRL-4和MOD间10个DEM的miRNA-Target调控网络,筛选出6个枢纽基因,它们被认为是治疗PF基因调控网络核心分子。SRL-4 对PF的改善作用可能与miR-433-3p、novel_202 和 miR-150-3p以及6个核心基因有关。嘌呤嘧啶代谢相关通路信号通路可能是治疗PF 的关键靶点和重要途径。
付新悦
,
宋欣妮
,
刘佳丽
,
刘玉键
,
石松利
,
钮树芳
,
常虹
,
王朋
,
齐君
,
白万富
. 蒙药扫日劳-4治疗肺纤维化大鼠的microRNAs比较及调控网络分析*[J]. 包头医学院学报, 2025
, 41(5)
: 6
-14
.
DOI: 10.16833/j.cnki.jbmc.2025.05.002
Objective: To investigate the impact of Mongolian medicine saorilao-4 (SRL-4) on miRNA and core genes involved in the gene regulatory network of lung tissue in rats with pulmonary fibrosis (PF). Methods: Rats were randomly divided into four groups: blank control group (CON), model group (MOD), positive drug control group, and SRL-4 group. Except for the CON group, rats in the other groups were administered with bleomycin via intratracheal injection to establish a pulmonary fibrosis model. The total RNA was extracted from rat lung tissue for transcriptome sequencing. The differentially expressed miRNA (DEM) in each group was screened by the difference analysis software edge R. The differentially expressed gene (DEG) of DEM was predicted by miRanda. The GO and KEGG were used to analyze the biological function enrichment of DEG. Cytoscape was used to construct the target gene regulatory network and screen the core genes. Results: Compared with CON, MOD selected 16 DEMs. Compared with MOD, SRL-4 screened 10 DEMs and regulated 63 052 target genes. GO analysis showed that the DEGs of SRL-4 and MOD were enriched in 52 GO entries. KEGG analysis showed that DEG was enriched in 182 signaling pathways, among which the number of genes related to purine metabolic pathway was the highest. By constructing a gene regulatory network, six core genes were screened, namely Spata25, Sultan1a1, Mpv17i, Cryba4, Jakmip3 and Fkbp5. Conclusion: By constructing a miRNA-Target regulatory network between SRL-4 and MOD, we identified six hub genes that are considered key molecules in the gene regulatory network for the treatment of PF. The improvement effect of SRL-4 on PF may be related to miR-433-3p, novel_202, miR-150-3p, and these 6 core genes. The purine metabolism-related signaling pathway may be a critical target and essential pathway for the treatment of PF.
[1] Raghu G, Remy-Jardin M, Myers JL, et al. Diagnosis of idiopathic pulmonary fibrosis. an official ATS/ERS/JRS/ALAT clinical practice guideline[J]. Am J Respir Crit Care Med, 2018, 198(5): e44-e68.
[2] Selman M, Pardo A. Revealing thepathogenic and aging-related mechanisms of the enigmatic idiopathic pulmonary fibrosis. An integral model[J]. Am J Respir Crit Care Med, 2014, 189: 1161-1172.
[3] Fingerlin TE, Murphy E, Zhang W, et al. Genome-wide association study identifies multiple susceptibility loci for pulmonary fibrosis[J]. Nat Genet, 2013, 45: 613-620.
[4] Yang S, Xie N, Cui H, et al. MiR-31 is a negative regulator of fibrogenesis and pulmonary fibrosis[J]. FASEB J, 2012, 26(9): 3790-3799.
[5] Friedman RC, Farh KK-H, Burge CB, Bartel DP. Most mammalian mRNAs are conserved targets of microRNAs[J]. Genome Res, 2009, 19(1): 92-105.
[6] 恒泰, 莲花, 阿木古楞. 简述蒙成药扫日劳-4汤[J]. 中国民族医药杂志, 2022, 28(4): 55-56.
[7] 白万富, 刘玉键, 李想, 等. 蒙药扫日劳-4味汤对特发性肺纤维化模型大鼠的改善作用及机制初探[J]. 中国药房, 2021, 32(12): 1435-1441.
[8] 陆松梅, 雷双奕, 李代蓉, 等. miR-433-3p通过靶向下调GALNT1调控肺癌细胞的生物学行为[J]. 中国细胞生物学学报, 2021, 43(11): 2126-2133.
[9] 郭义威, 孙春莉, 王树兴. miR-433在NIH-3T3细胞纤维化和矽尘诱导的小鼠肺纤维化中的作用[J]. 中国病理生理杂志, 2020, 36(8): 1458-1464.
[10] Ghédir H, Braham A, Viville S, et al. Comparison of sperm morphology and nuclear sperm quality in SPATA16- and DPY19L2-mutated globozoospermic patients[J]. Andrologia, 2019, 51(6): e13277.
[11] Dammanahalli JK, Duffel MW. Oxidative modification of rat sulfotransferase 1A1 activity in hepatic tissue slices correlates with effects on the purified enzyme[J]. Drug Metab Dispos, 2012, 40(2): 298-303.
[12] Billingsley G, Santhiya ST, Paterson AD, et al. CRYBA4, a novel human cataract gene, is also involved in microphthalmia[J]. Am J Hum Genet, 2006, 79(4): 702-709.
[13] Zhou J, Jiang G, Xu E, et al. Identification of SRXN1 and KRT6A as key genes in smoking-related non-small-cell lung cancer through bioinformatics and functional analyses[J]. Front Oncol, 2022, 11: 810301.
[14] Sun L, Gang X, Li Z, et al. Advances in understanding the roles of CD244 (SLAMF4) in immune regulation and associated diseases[J]. Front Immunol, 2021, 12: 648182.
[15] 徐玉雪. FKBP51在小鼠肝脏纤维化中的作用以及分子机制的探究[D]. 北京: 北京协和医学院(清华大学医学部)&中国医学科学院, 2016.
[16] 李文超. miRNA调控纳米SiO2诱导肺损伤靶基因作用的初步研究[D]. 江苏: 东南大学, 2015.
[17] Lao C , Li W , Li M ,et al.The biological effect of MiR-541 in rat′s pulmonary fibrosis induced by nanosized SiO2 through targeting TGF-β RIII[J]. Nanomedicine Nanotechnology Biology & Medicine, 2016, 12(2): 563-563.
[18] 蔡萧君, 王钦, 江柏华, 等. 基于粪便代谢组学技术丹贝益肺方对肺纤维化伴抑郁大鼠的作用机制研究[J]. 山西医科大学学报, 2022, 53(2): 175-181.
[19] Jordheim LP, Peters GJ. New insights in research on purine and pyrimidine metabolism[J]. Nucleosides Nucleotides Nucleic Acids, 2022, 41(3): 247-254.
[20] Bueno LCM, Paim LR, Minin EOZ, et al. Increased serum mir-150-3p expression is associated with radiological lung injury improvement in patients with COVID-19[J]. Viruses, 2022, 14(7): 1363.
