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AIM: To investigate the mechanism and reversal effect of Zuo Jin Wan (ZJW) on cetuximab (CET) resistance in KRAS mutant colorectal cancer cell. METHODS: The mutation status of KRAS gene in SW620, Lovo, HCT116, HT29 and Caco2 cells were detected by Sanger sequencing. CCK-8 assay was used to detect the effects of ZJW, CET, ZJW combined with CET and CET, ZJW in combination with other cell death inhibitors on the survival rate of the above cells, and to observe the reversal effects of ZJW on CET-treated KRAS mutant cells (SW620, Lovo and HCT116). Flow cytometry, colorimetric method, and Fe
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Objective:To analyze the possible mechanism of Zuojin Pills on gastroesophageal reflux disease based on network pharmacology. Methods:By searching for the active constituent and protein targets of Zuojin Pills in TCMSP database,the protein names were converted into gene names in Uniprot database. Cytoscape 3.7.1 was used to draw the active constituent-target-medicine network diagram of Zuojin Pills and analyze the topological parameters. Then find the target of gastroesophageal reflux disease through OMIM,GeneCards,DRUGBANK database, find the intersection target of medicine and disease, perform PPI network analysis on the intersection target in STRING 11.0, and use the Metascape database to enrich the intersection target for further analysis. Cytoscape 3.7.1 was used to draw a network diagram of the active constituent- target-pathway of the medicine and to conduct a topology parameter analysis. Results:The main active constituent of Zuojin Pills in the treatment of gastroesophageal reflux disease are quercetin, Evodiamine, R-tetrahydroberberine, 1-methyl-2-nonyl-4-quinolone, berberine, etc. Targets include PTGS2, NOS3, MAPK1, EGFR, TNF, IL6, ERBB2, VEGFA, EGF, IL1B, etc., and these processes are mainly completed through inflammatory response, cancer, cell proliferation and apoptosis, cell connection, etc. Conclusions:The treatment of gastroesophageal reflux disease with Zuojin Pills is a complex process with multiple constituent, multiple targets, and multiple pathways. It is hoped that it which could provide reference for the future research on its mechanism of action.
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Objective:To investigate the effect of Zuojin Pill on the expression of hypoxia inducible factor-1α (HIF-1α) and apoptosis of thyroid cells in rats with autoimmune thyroiditis (AIT). Methods:Ninetysix rats were divided into control group, model group, standarlized protocol and low, medium and high dose Zuojin Pill groups. Except the control group, the rats in other groups were produced as AIT model. After successful modeling, rats in low-, medium-, and high-dose groups were intragastrically administered with 0.63, 1.26, and 2.52 g/kg Zuojin Pill respectively, standarlized protocol was given 6.25 mg/kg Tripterygium wilfordii polyglycoside, and the control group and model group were given the same amount of normal saline. The levels of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) were detected by enzyme-linked immunosorbent assay (ELISA); the activity of SOD and the content of MDA in thyroid tissue were detected by kit; the pathological changes of thyroid tissue were observed by HE staining; the apoptosis of thyroid cells was detected by TUNEL; the expression of HIF-1α, B-cell lymphoma-2 (Bcl-2), Bcl-2-related X protein (Bax) and Cleaved caspase-3 in thyroid tissues was detected by Western blot. Results:Compared with the model group, the content of TNF-α, TPOAb and TGAb in serum of rats in the low, medium and high dose Zuojin Pill groups significantly decreased, the content of IL-10 in serum significantly increased ( P<0.05). The SOD activity significantly increased, and the MDA content significantly decreased in the low, medium and high dose Zuojin Pill groups ( P<0.05). The apoptosis rate significantly decreased in the low, medium and high dose Zuojin Pill groups ( P<0.05). The expression of HIF-1α (0.48 ± 0.05, 0.63 ± 0.06, 0.86 ± 0.06 vs. 0.33 ± 0.03), Bcl-2 (0.48 ± 0.04, 0.59 ± 0.05, 0.68 ± 0.04 vs. 0.37 ± 0.04) significantly increased, the expression of Bax (0.67 ± 0.05, 0.53 ± 0.05, 0.40 ± 0.05 vs. 0.80 ± 0.06), Cleaved caspase-3 (0.64 ± 0.06, 0.51 ± 0.03, 0.36 ± 0.03 vs. 0.77 ± 0.05) significantly decreased in the low, medium and high dose Zuojin Pill groups ( P<0.05). Conclusion:Zuojin Pill could enhance the expression of HIF-1α, regulate the secretion of inflammatory factors, reduce the level of oxidative stress and thyroid autoantibody, and inhibit the apoptosis of thyroid cells.