健脾理气方大鼠含药血清对体外人肝癌细胞株（HepG2）作用的实验研究

【作者】 程学莲；

【导师】 王伯庆；

【作者基本信息】 辽宁中医药大学 ， 中医基础理论， 2008， 硕士

【摘要】 目的:观察健脾理气方对人肝癌细胞株(HepG2)的增殖抑制和细胞凋亡诱导作用。方法:以血清药理学方法,将健脾理气方含药大鼠血清用于HepG2,采用MTT法检测其效果及量效关系,观察健脾理气方对细胞增殖的影响;酶细胞化学染色法观察HepG2酶活性的改变;光镜观察在HE染色、免疫组化条件下HepG2的P53、Bcl-2的蛋白表达。结果:MTT法检测显示,健脾理气方组在48h后,对HepG2的抑制率可达32.86%,顺铂组在48h后其抑制率可达50%,健脾理气方加顺铂组在48h后抑制率即可超过50%;酶细胞化学染色法检测显示HepG2的Mg2+-ATP酶、G-6-P酶活性降低;用药各组P53蛋白表达有不同程度的增强、Bcl-2基因蛋白表达也有不同程度的降低。结论:健脾理气方含药血清具有抑制HepG2的增殖和诱导凋亡的作用,与顺铂合用有协同作用,其机制可能与降低HepG2的代谢功能,以及促使P53基因表达增强和抑制基因Bcl-2表达下降过程相关。更多还原

【Abstract】 Object:To observe the inhibit cell ptoliferation and induce apoptosis function of Jianpiliqi formula(JLF)to Person’s liver cancer cell HepG2.Methods:Using serum pharmacological method to culture human liver cancer cell line HepG2,the relation of concentration-effect was detected by MTT,to observe the cell proliferation inhibited.Enzyme cytochenminstry was used to detect the activity of G-6-Pase and Mg2+-ATPase of the liver cancer cell.The effect of Jianpiliqi formula(JLF)contained serum on cell expressions of Bcl-2 and P53 were observed by lens and immunocytochemistry.Results:After treated with Jianpiliqiformula contained serum,MTT tests showed the relation of concentration-effect,After 48h,the cell proliferation inhibited rate of Jianpiliq formula is 32.86%,about 50% of DDP group,over 50% of JLF+DDP group.Enzyme cytochemistry method showed that the response granules of G-6-Pase and Mg2+-ATPase in the human liver cancer cell line HepG2 become smaller afrer the treatment of serum container Chinese herbs,the amount of the granules became feweer,the density was lower.Immunocytochemistry indicated that JLF can increase the expression of anti-oncogene P53,and at the same time ,the protein expression of Bcl-2 for depressing apoptosis decreased.Conclusion:The activity of the G-6-Pase and Mg2+-ATPase decreased after using the Jianpiliqiformula contained serum.The HepG2 cells were inhibited ptoliferation and induced to apoptosis by the pharmacy serums of Jianpiliqi formula,prominently by the granules associating with cisplatin.The mechanism of JLF in anti-tumor might be reduce the human hepatoma cells (HepG2) physiological function and achieved through inducing apoptosis and work on the related gene expression of Bcl-2 and P53.更多还原