[Molecular mechanism of Xiangsha Liujunzi Decoction in treating chronic atrophic gastritis based on transcriptome...

Based on transcriptomics technology, this study investigated the molecular mechanisms of Xiangsha Liujunzi Decoction in treating chronic atrophic gastritis(CAG), which were confirmed through experimental validation. The CAG rat model was built by the MNNG composite multi-factor method, followed by a 90-day administration of Xiangsha Liujunzi Decoction. The study measured the rat body mass and 3-hour food intake in each group and observed the pathological changes in gastric tissue using HE staining. It used RNA sequencing(RNA-Seq) to determine the gene expression in the gastric tissue of rats in each group and then analyzed differential genes through KEGG enrichment analysis to validate enriched key signaling pathways. The results indicated that the body mass and 3-hour food intake in the model group were significantly lower than those in the blank group, alongside typical signs of gastric tissue atrophy. In contrast, the treatment group exhibited a significant increase in body mass and 3-hour food intake and significant improvements in the damage of gastric tissue pathology. A total of 944 genes associated with the improvement of CAG by Xiangsha Liujunzi Decoction were obtained via RNA-Seq. KEGG enrichment analysis revealed that these genes were mainly enriched in multiple signal transduction pathways involving signaling pathways such as inflammation, proliferation, apoptosis, as well as cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate(cGMP)/protein kinases(PKG) pathways. Compared to the blank group, the RNA-Seq results showed significant increases in adenosine A1 receptor recombina(Adora1) and BK channel subunit beta 3(Kcnmb2) mRNA expression in gastric tissue in the model group, while the expression of endothelin receptor type B gene(Ednrb), recombinant Rho associated coiled coil containing protein kinase 2(Rock2), cGMP-dependent protein kinase 2(Prkg2), and cAMP responsive element binding protein 3 like 3 gene(Creb3l3) mRNA was significantly decreased. Compared to the model group, the expression of Adora1 and Kcnmb2 mRNA in gastric tissue in the treatment group decreased significantly, while the expression of Ednrb, Rock2, Prkg2, and Creb3l3 mRNA increased significantly. Molecular biology experiments revealed significant decreases in cAMP content in gastric tissue and plasma, as well as decreased expression levels of Ras homolog gene family member A(RhoA), ROCK2, phospho-myosin phosphatase target subunit 1(p-MYPT1), and phospho-Myosin light chain(Ser20)(p-MLC20) proteins in gastric tissue in the model group, accompanied by significantly increased cGMP content in gastric tissue and plasma and significantly increased expression of PKG protein in gastric tissue. When the treatment group was compared with the model group, the results showed significant decreases in cGMP content in gastric tissue and plasma and PKG protein expression in gastric tissue, alongside increased expression le-vels of RhoA, ROCK2, p-MYPT1, and p-MLC20 in gastric tissue. At the same time, the results of RT-qPCR confirmed that the expression of key genes in the cGMP/PKG pathway was consistent with the RNA-Seq findings. In summary, Xiangsha Liujunzi Decoction demonstrates significant therapeutic efficacy in CAG rats, and the specific mechanism may be through the abnormal expression of signaling molecules that regulate the cGMP/PKG pathway and downstream processes.