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目的研究0.05g·L~(-1)～3.5g·1~(-1)浓度苦参碱对人肝癌细胞株HepG2的增殖抑制及其剂量与抑制方式的关系,为进一步研究苦参碱抑制HepG2细胞增殖的机制打下基础;为临床应用苦参碱治疗肿瘤奠定基础。方法苦参碱处理细胞后,MTT法、细胞计数法、H~3-TdR掺入法测定HetG2细胞存活率、细胞生长曲线、DNA合成率的变化;台盼蓝拒染法和细胞外液LDH活性测定观察细胞膜的完整性。结果苦参碱浓度小于0.2g·L~1时不能抑制HepG2的增殖,细胞存活率大于80％;0.7g·L~(-1)～0.8g·L~1时,HepG2对药物中度敏感,细胞存活率为30％～50％:大于1.0g·L~1时为抗HepG2的敏感药物浓度,细胞存活率小于30％;苦参碱大于3.0g·L~(-1)时,细胞膜完整性破坏,以坏死细胞为主。而苦参碱对大鼠肝细胞影响很小,达1.5g·L~(-1)时,细胞存活率仍为72.4％。结论一定浓度苦参碱可抑制HepG2细胞增殖并且抑制作用呈时间和剂量依赖性。0.8g·L~(-1)可作为诱导HepG2细胞分化的作用浓度;1.5g·L~(-1)～2.5g·L~(-1)可作为诱导HepG2凋亡的作用浓度;苦参碱大于3.0g·L~(-1)时主要引起HepG2坏死。
Objective To investigate the inhibitory effect of matrine on the proliferation of human hepatocellular carcinoma cell line HepG2 at a concentration of 0.05g·L -1 to 3.5g·1 -1 and the relationship between the dosage and the mode of inhibition to further study the matrine inhibition. The underlying mechanism of HepG2 cell proliferation lays the foundation for the clinical application of matrine in the treatment of tumors. Methods After treated with matrine, MTT assay, cell counting, and H~3-TdR incorporation method were used to determine the survival rate, cell growth curve and DNA synthesis rate of HetG2 cells; trypan blue exclusion and extracellular LDH. The activity assay measures the integrity of the cell membrane. Results When matrine concentration was less than 0.2g·L~1, the proliferation of HepG2 could not be inhibited and the cell survival rate was greater than 80%. When the concentration of matrine was 0.7g·L~(-1) to 0.8g·L~1, HepG2 was moderately sensitive to drugs. The cell survival rate is 30% to 50%: When the concentration is greater than 1.0 g·L~1, the concentration of the anti-HepG2 drug is sensitive, the cell survival rate is less than 30%; when the matrine is greater than 3.0 g·L -1, the cell membrane Integrity is disrupted by necrotic cells. Matrine has little effect on rat hepatocytes, and the survival rate of the cells is still 72.4% when reaching 1.5 g·L -1 . Conclusion A certain concentration of matrine can inhibit the proliferation of HepG2 cells in a dose- and time-dependent manner. 0.8g·L~(-1) can be used as a concentration to induce HepG2 cell differentiation; 1.5g·L~(-1) to 2.5g·L~(-1) can be used to induce the apoptosis of HepG2; When the base is greater than 3.0 g·L -1 , HepG2 is mainly caused to necrosis.