知识与财富的链接
目的：基于微小核糖核酸(miRNA)测序探究理中汤对胃溃疡(GU)脾胃虚寒证大鼠胃黏膜的修复效应与作用机制。方法：60只SD大鼠随机分为正常组、模型组、阳性药组及理中汤低、高剂量组。通过高通量测序筛选差异表达miRNA,进行基因本体(GO)和京都基因与基因组百科全书(KEGG)分析。采用苏木精-伊红(HE)染色、电镜观察胃黏膜形态,酶联免疫吸附测定法检测胃蛋白酶及氧化应激指标,实时荧光定量PCR检测miRNA表达。结果：高通量测序发现10个核心差异表达miRNA,其中4个表达下调,6个表达上调;GO和KEGG分析显示脾胃虚寒型GU发病与细胞死亡、代谢、节律、炎症等通路相关。与正常组比较,模型组出现典型脾胃虚寒证候及胃黏膜损伤;阳性药组及理中汤高剂量组大鼠脾胃虚寒证候明显改善,溃疡指数降低,胃蛋白酶活性下降,氧化应激指标改善,胃组织病理损伤和超微结构亦显著改善,miR-539-5p、miR-PC-5p-51319＿24等6个miRNA表达上调,miR-24b-3p、miR-PC-5p-44136＿35等4个miRNA表达下调(均P<0.05)。结论：理中汤能促进GU脾胃虚寒证大鼠胃黏膜修复,减轻氧化应激反应,其作用机制可能与调控与脾胃虚寒证GU发病密切相关的miRNA表达有关。
Based on microRNA（miRNA) sequencing,this study investigated the reparative effects and mechanisms of Lizhong Decoction on gastric mucosa in rats with gastric ulcer（GU) of spleen-stomach deficiency-cold syndrome.Methods:Sixty Sprague Dawley rats were randomly divided into a normal group,a model group,a positive drug group,and low-and high-dose Lizhong Decoction groups.Differentially expressed miRNAs were screened by high-throughput sequencing and analyzed using Gene Ontology（GO) and Kyoto Encyclopedia of Genes and Genomes（KEGG).Gastric mucosal morphology was assessed by hematoxylin-eosin（HE) staining and electron microscopy.Gastric pepsin activity and oxidative stress markers were measured by enzyme-linked immunosorbent assay（ELISA),and miRNA expression levels were detected by real-time fluorescent quantitative PCR（qRT-PCR).Results:High-throughput sequencing identified 10 core differentially expressed miRNAs,of which 4 were downregulated and 6 were upregulated.GO and KEGG analyses indicated that the pathogenesis of spleen-stomach deficiency-cold type GU is associated with pathways related to cell death,metabolism,circadian rhythm,and inflammation.Compared with the normal group,the model group exhibited typical manifestations of spleen-stomach deficiency and cold and gastric mucosal damage.In the positive drug and high-dose Lizhong Decoction groups,these manifestations were significantly improved.Specifically,ulcer indices were reduced,gastric pepsin activity decreased,oxidative stress markers were ameliorated,and histopathological and ultrastructural damages of gastric tissues were substantially alleviated.The expression of six miRNAs,including miR-539-5p and miR-PC-5p-51319_24,was significantly upregulated,while four miRNAs,including miR-24b-3p and miR-PC-5p-44136_35,were significantly downregulated（all P<0.05).Conclusion:Lizhong Decoction promotes gastric mucosal repair in rats with GU of spleen-stomach deficiency-cold syndrome,alleviates oxidative stress,and its mechanism may be closely related to the regulation of miRNA expression associated with the pathogenesis of GU in this syndrome.
