墓头回环烯醚萜酯提取部位抗肿瘤作用及机制研究

Antitumor effect of iridoid ester fraction from Patriniae Radix and its mechanism

目的研究墓头回醋酸乙酯提取物环烯醚萜酯部位（PHEBB）的体内外抗肿瘤作用及其机制。方法硅胶柱色谱法精制PHEBB；反相高效液相色谱法分析PHEBB化学成分。MTT法检测PHEBB对人肿瘤细胞KB、COLO-205、SGC-7901的抑制活性。采用小鼠移植性瘤模型研究PHEBB对小鼠S180 肉瘤、H22肝癌的体内抗肿瘤作用；免疫组化法研究PHEBB对肿瘤组织中相关基因表达的影响。结果PHEBB主要成分均为环烯醚萜酯类，主要有desacetylisovaltratum、isovaltrateacetoxyhydrin。体外实验结果显示，PHEBB对KB、COLO-205、SGC-7901等癌细胞均具有显著抑制作用，Icsn在2．27～5．49μg／mL。体内实验显示，PHEBB以60mg／kgip给予荷瘤小鼠，对S180肉瘤的抑制率为45．5％，对H22肝癌的抑制率为54．2％；免疫组化检测结果显示，PHEBB能上调小鼠H22肿瘤组织中Bax表达，下调Bcl-2表达，且能降低肿瘤组织中微血管密度。结论PHEBB抗肿瘤药效物质为环烯醚萜酯；其体内外均具有一定抗肿瘤作用，且其抗肿瘤机制可能与诱导肿瘤细胞凋亡及抑制肿瘤微血管有关。

Objective To purify the iridoid ester fraction of ethyl acetate extract from Patriniae Radix （PHEBB）, and to study the antitumor effect and mechanism in vivo and in vitro. Methods The silica gel column chromatography was used to purify the effective fraction. The chemical constituents of PHEBB were elucidated by RP-HPLC; MTT assay was used to study the inhibition of PHEBB on human oral epithelial cancer cells KB, human colon cancer cells COLO-255, and human gastric cancer cells SGC-7901; Mice transplanted S18o and H22 experiments were used to study the antitumor effect in vivo; The immunohistochemical method was used to study the effect of IEEAM on the expression of related genes in tumor tissue. Results The main effective components in PHEBB were desacetylisovaltratum and isovaltrate acetoxyhydrin, and they are both iridoid esters. The results of MTT showed that PHEBB had the significant inhibition on KB, COLO-205, and SGC-7901 cells, and the IC50 values were within 2.27--5.49 μg/mL; The experiment of mice transplanted tumor showed that after ip injection with PHEBB at a dose of 60 mg/kg, there was an inhibitory rate of 45.5% on S180 and 54.2% on H22; The immunohistochemistry results showed that PHEBB could upregulate Bax expression in H22 cells, decrease Bcl-2 expression, and reduce the microvascular density in tumor tissue. Conelusion The main components in PHEBB are iridoid esters, and have certain antitumor effect in vitro and in vivo; the antitumor mechanism of PHEBB might be associated with the induction of apoptosis and the inhibition of tumor microvessel.