Study on the mechanism of Zhenggan Decoction in the treatment of liver cancer based on mitophagy
Objective:To investigate the mechanism by which Zhenggan formula regulates mitophagy to inhibit the growth of H22 hepatocellular carcinoma (HCC)xenografts in mice.Methods:A mouse model of H22 HCC xenografts was established and randomly divided into six groups:blank,model,low-dose Zhenggan formula,medium-dose Zhenggan formula,high-dose Zhenggan formula,and sorafenib. The treatment was administered for 14 days.The mice were euthanized 12 hours after the last administration.Tumors,thymus,and spleens were excised,weighed,and recorded.Tumor inhibition rate,thymus index,and spleen index were subsequently calculated.HE staining was used to observe tumor tissue structure;TUNEL staining was performed to assess apoptosis in tumor cells.ELISA was used to measure serum AFP levels.Transmission electron microscopy was employed to observe changes in mitochondrial ultrastructure and levels of mitophagy in tumor tissues,and immunohistochemistry was conducted to detect the expression of autophagy-related proteins LC3 and P62.Results:Compared to the model group,the high-dose Zhenggan formula significantly increased the body weight of HCC mice (P<0.05 )and effectively reduced the tumor weight (P<0.01 ).HE staining indicated a reduction in tumor cells,membrane rupture,nuclear fragmentation,and necrotic areas of varying sizes in the Zhenggan formula groups.The apoptosis level of liver cancer cells in all drug-treated groups increased significantly (P<0.01),accompanied by a notable decrease in serum AFP levels (P<0.01).The therapeutic efficacy of the high-dose group was comparable to that of the sorafenib group.The apoptosis level of liver cancer cells in all drug-treated groups increased significantly (P<0.01 ),and a notable decrease in serum AFP levels (P<0.01 ).The therapeutic efficacy of the high-dose group was comparable to that of the sorafenib group.Transmission electron microscopy showed mitochondrial swelling,cristae fracture,disordered cristae structure,vacuoles,and autophagosomes in the ZGF-treated tumor tissues.Immunohistochemistry results demonstrated an increased LC3 protein expression and a decreased P62 protein expression,demonstrating a dose-dependent trend.Conclusion:Zhenggan formula effectively inhibits the growth of H22 hepatocellular carcinoma xenografts.It elevates the expression of the autophagy protein LC3,reduces P62 expression,and increases autophagosome formation,suggesting that its mechanism may be related to the activation of mitophagy.
万方数据
维普
