基于红细胞免疫功能调节的石蒜碱体内抗肿瘤作用机制研究

目的 研究石蒜碱对肝癌H22细胞荷瘤小鼠红细胞膜结构功能的影响.方法 通过抑瘤率实验、生命延长率实验、流式细胞仪检测肿瘤细胞凋亡率实验,验证石蒜碱的体内抗肿瘤活性;采用试剂盒测定红细胞膜总蛋白、胆固醇和唾液酸含量,考察石蒜碱对H22荷瘤小鼠红细胞膜主要组分的影响;采用荧光偏振法检测红细胞膜流动性,荧光染色法检测红细胞膜...

Anti-tumor mechanism of lycoline in vivo based on regulation of red blood cell immune function

ObjectiveTo study the effect of lycorine on structure and function of red blood cell membranes in H₂₂ tumor-bearing mice.Methods Anti-tumor activity of lycorine in vivo was verified by tumor inhibition rate experiment, life extension rate experiment, and tumor cells apoptosis rate test by flow cytometry. Contents of total protein, cholesterol and sialic acid of erythrocyte membrane were determined by assay kit to detect the effect of lycorine on content of main components of erythrocyte membrane in H₂₂ tumor-bearing mice. The erythrocyte membrane fluidity was detected by fluorescence polarization method, and erythrocyte membrane sealing degree was determined by Zamudio method to verify the effect of lycorine on erythrocyte membrane structure function of H₂₂ tumor-bearing mice. Adenosine triphosphate (ATP) activity assay kit, fluorescence spectrophotometer, laser confocal scanning microscopy and Western blotting method were used to detect the effect of lycorine on erythrocyte membrane ion channel activity of H₂₂ tumor-bearing mice. Western blotting was used to detect the effects of lycorine on protein expressions of cleaved Caspase-3, cleaved Caspase-9, B-cell lymphoma 2 (Bcl-2) and Bcl-2 associated X protein (Bax) in tumor cells of H₂₂ tumor-bearing mice. ResultsLycorine inhibited tumor growth in H₂₂ tumor-bearing mice, prolonged the survival time of H₂₂ tumor-bearing mice, and induced tumor cell apoptosis in H₂₂ tumor-bearing mice (P < 0.05, 0.01); Lycorine increased erythrocyte membrane total protein, sialic acid and cholesterol contents in H₂₂ tumor-bearing mice, improved the fluidity and tightness of erythrocyte membrane, enhanced the erythrocyte membrane Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity of H₂₂ tumor-bearing mice, increased the intracellular pH of H₂₂ tumor-bearing mice, decreased H⁺, Ca²⁺, Cl⁻ concentrations in red blood cells, enhanced the HCO³⁻/Cl⁻ transport activity of H₂₂ tumor-bearing mice, and increased Band 3 protein expression (P < 0.01). Lycorine increased the protein expressions of cleaved Caspase-3 and cleaved Caspase-9, increased the expression of pro-apoptotic protein Bax, decreased the expression of anti-apoptotic protein Bcl-2, and reduced the ratio of Bcl-2/Bax (P < 0.01). ConclusionLycorine has anti-tumor effect on H₂₂tumor-bearing mice, and can improve the activity of cell membrane ion channel by changing the erythrocyte membrane structure of H₂₂ tumor-bearing mice, and establish a relationship between ion channels to promote each other. Through this mutual promotion, the intracellular pH and ion balance are adjusted, and the structural function of erythrocyte membrane is improved, thereby enhancing the immune function of red blood cells. In addition, lycorine regulates the activity of apoptosis-related proteins in tumor cells, activates the apoptosis-related factors, initiates the apoptosis-related process of tumor cells, and exerts the anti-tumor effect.