Investigating the Effect of Hydrogen-Rich Water on Liver Cell Injury and Liver Cancer by Regulating GP73/ TGF-β Pathway
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Abstract:Background Liver cancer is an extremely heterogeneous malignant disease among tumors identified to date. In recent years, a large number of studies have found that low-concentration hydrogen or hydrogen-rich water or hydrogen-saturated physiological saline has a protective effect on many diseases. Observing the intervention effect of hydrogen-rich water and speculate whether its effect is regulated by GP73/TGF-β signaling pathway would provide a basis for clinical treatment using hydrogen in liver cancer.Method The(N-nitrosodiethylamine) DEN-induced LX-2 hepatocyte injury model, hepatocarcinoma cell(HepG2 cells), and normal hepatic stellate cell LX-2 and HepG2 co-culture system were treated with hydrogen-rich water, and the cell viability was measured by the Cell Counting Kit-8(CCK-8)method. Also, the supernatant was collected to determination the transforming growth factor (TGF-β), tumor necrosis factor (TNF-α), and osteopontin (OPN) content. Subsequently, qPCR method was employed to detect the expression of GP73, TGF-β, and Smad2 mRNA in the cell. Total cell protein was extracted, and the level of GP73, TGF-β, and Smad2 proteins in the cells was detected by Western blot.ResultAfter the intervention of hydrogen-rich water, DEN increases the activity of LX-2 hepatocytes, inhibits the secretion of TGF-β, downregulate the expression of GP73, TGF-β, and Smad2 mRNA in the injured LX-2 cells, inhibits the expression of GP73 and TGF-β proteins, and exerts a protective effect on injured cells. The intervention with hydrogen-rich water in HepG2 liver cancer cells inhibits the expression of GP73 mRNA and protein in the cells, upregulates the expression of TGF-β mRNA and protein, inhibits the expression of OPN, and exerts an anti-proliferation effect on liver cancer cells. In the co-culture system of LX-2 and HepG2, the effect of hydrogen-rich water showed different effects with the altered ratio of the two cells in the system.Conclusion Hydrogen-rich water has a protective effect on liver cell damage and an inhibitory effect on liver cancer cells, which is effectuated by regulating the GP73/TGF-β signaling pathway.