Huazhuojiedu Decoction Regulates Mitophagy to Alleviate Precancerous Lesions of Gastric Cancer via Sirt3/Foxo3a/Parkin Pathway

Abstract

Abstract Backgroud：Huazhuojiedu decoction (HZJD) has been demonstrated to be effective in the treatment of precancerous lesions of gastric cancer (PLGC). We aimed to explore the potential mechanisms of HZJD for alleviating PLGC in vivo and in vitro. Methods: The PLGC rat model was established by administrating 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG) and sodium for 24 weeks, followed by 10 weeks of HZJD decoction or vitamin B12 therapy. The PLGC cell model (MC) was prepared by inducing human gastric mucosal epithelial cells (GES-1) with MNNG. HZJD decoction and vitamin B12 drug-containing serum were given to treat MC cells, meanwhile sirt3 siRNA was transfected into MC cells. The CCK-8 assay and the EdU assay were used to detect cell proliferation. The histopathological changes of gastric tissues were observed by H&E staining, HID/AB staining and AB/PAS staining. The mRNA and protein expressions of on mitophagy-related molecules were detected by RT-qPCR and Western blot. Immunohistochemistry was used to test the differential expressions of sirt3/foxo3a/parkin pathway. Immunofluorescence was used to evaluate mitophagic level. Transmission electron microscopy was used to monitor the degree of mitochondrial damage and the occurrence of mitophagy. Results: The results indicated that HZJD could retard the pathological progress of gastric mucosa in PLGC rats and reduce the elevated cell proliferation in MC cells. The treatment of HZJD could significantly increase the gene and protein expressions of sirt3, foxo3a, parkin, LC3Ⅱ/Ⅰ, meanwhile decrease the mRNA and protein expressions of p62, tomm20. The colocalization of LC3 and COX Ⅳ was inhibited in PLGC rats, besides the fluorescent intensity of mitophagy was weakened in MC cells. This downtrend of mitophagic level in vivo and in vitro could be reversed by HZJD. More importantly, the improvement of mitophagy by HZJD was associated with sirt3/foxo3a/parkin pathway. Conclusions: Our results suggested that HZJD decoction could ameliorate PLGC in vivo and in vitro, and its therapeutic effect might be related to regulating mitophagy via sirt3/foxo3a/parkin pathway.