Objective

2

To explore the potential mechanism of

Xianglian Huazhuo Formula

（香连化浊方， XHF） in the treatment of chronic atrophic gastritis （CAG）.

Methods

2

The gene chip dataset was screened out by using the GEO database， and the CAG proliferation-related differential genes were obtained based on the GEO2R tool and GeneCards database. The Metascape platform was used to perform GO and KEGG enrichment of CAG proli-feration-related differential genes so as to find biological processes and important signaling pathways related to CAG， and conduct animal experiments to verify. From 70 Wistar rats， 14 rats were randomly selected as the normal group for routine feeding， and the remaining 56 rats were given “N-methyl-N'-nitro-nitrosoguanidine plus disordered feeding plus sodium salicylate” to prepare the CAG model. The 50 successfully modeled rats were randomly divided into model group， morodan group， and XHF high-， medium- and low-dose groups， with 10 rats in each group. The rats in the morodan group were given 1.4 g/（kg·d） of morodan solution by gavage， while those in the high-， medium- and low-dose XHF groups were given 36， 18 and 9 g/（kg·d） of XHF solution by gavage， respectively， and the rats in the normal group and the model group were given 0.01 ml/g of normal saline by gavage， once a day， for 60 consecutive days. HE was used to observe the histopathological changes of the gastric mucosa of rats， and transmitted electron microscopy was taken to observe the ultrastructure of gastric mucosal cells.The protein expression levels of epidermal growth factor receptor （EGFR）， phosphorylated extracellular signal-regulated protein kinase 1 （p-ERK1） and proto-oncogene （c-myc） were detected by Western-blot， and the mRNA expression levels of EGFR， extracellular signal-regulated protein kinase 1 （EPK1） and c-myc were detected by RT-PCR method.

Results

2

Finally， 45 CAG proliferation-related differential genes were obtained. GO and KEGG enrichment analysis showed that it could affect the positive regulation of cytokine production， positive regulation of mitogen-activated protein kinase （MAPK） cascade， the regulation of establishment of protein localization， involving multiple signaling pathways such as cancer pathway， MAPK signaling pathway， and helper T cell 17 （Th17） cell differentiation. HE staining showed that the thickness of gastric mucosa， the arrangement of lamina propria， and the infiltration of inflammatory cells in each medication group were improved compared to the model group， especially the high- and medium-dose XHF groups. The transmission electron microscopy showed that compared to those in the model group， the mitochondrial structure was restored， and the swelling of the endoplasmic reticulum was reduced in the medication groups， and more improvements were seen in the high- and medium-dose XHF groups. Compared to the normal group， the model group had increased protein and mRNA expression levels of EGFR， p-ERK1 and c-myc in the gastric mucosa （

P

＜0.05）. Compared to those in the model group， the protein expression levels of EGFR and c-myc in the high-dose XHF group， c-myc in the morodan group， p-ERK1 in all medication groups， as well as mRNA expression levels of EGFR， ERK1 and c-myc in all medication groups significantly decreased （

P

＜0.05）. Compared to those in the morodan group， c-myc protein expression in the XHF low-dose group significantly increased， while the EGFR and c-myc mRNA expression levels in the XHF high- and medium-dose groups decreased， also the ERK1 mRNA expression level in the high- and low-dose XHF groups decreased （

P

＜0.05）.

Conclusion

2

XHF in the treatment of CAG may be related to the regulation of EGFR/MAPK/ERK signaling pathway， the blocking of abnormal differentiation and proliferation of gastric mucosal cells， and the reduction of gastric mucosal tissue damage in CAG.