Guo Ming, Wu Zhou-Ling, Wang Chun-Ge, Gao Xiao-Yan
Yao Xue Xue Bao. 2014 Mar;49(3):337-45.
Chelating ligand method has been used to synthesize baicalin-metal (Ni2+, Co2+, Cu2+) complexes (BMC). The composition and structure of BMC were characterized by the element analysis, ultraviolet spectrum (UV), infrared spectrum (IR), mass (MS) and thermal gravitational analysis (TGA). MTT was used to analyze the effects of BMC on SMMC-7721 cell proliferation. PI staining method and Annexin-V/FITC double staining method were used to analyze the effects of BMC on the cell cycle and apoptosis of SMMC-7721 cell. Fluorescence quantitative RT-PCR was used to analyze the expression of BMC on Bcl-2 gene and Bax mRNA, flow cytometry was used to analyze BMC on the expression of Bcl-2 protein and Bax protein. The antineoplastic activity and mechanism of action of BMC was explored comprehensively. The results showed that three new kinds of BMC (molar ratio of 2 : 1) were successfully prepared, the complexes molecular formula are: Na2Ni(C21H16O11)2 x 10H2O, Na2Co(C21H16O11)2 x 8H2O and Na2Cu(C21H16O11)2 x 8H2O. According to the results of cell cycle and apoptosis detection, BMC stopped cells at G0/G1 phase to S phase and G2/M phase. Gene and protein detection showed that under the given concentration and time, BMC can downregulate the expression of Bcl-2 gene in SMMC-7721 cells, and significantly decrease the expression of Bcl-2 protein, at the same time, with the increase of expression of Bax gene, the Bax protein's expression increased significantly. Which indicates that BMC restrain cell proliferation and cell apoptosis by stopping cell cycle, reducing the expression of Bcl-2 and increasing that of Bax; The anti-tumor activities of three kinds of complexes were: baicalin-copper (BC-Cu) > baicalin-cobalt (BC-Co) > baicalin-nickel (BC-Ni) > baicalin (BC), showing the dose-response relationship.
采用螯合配体法合成了黄芩苷 - 金属（Ni2 +、Co2 +、Cu2 +）配合物（BMC）。通过元素分析、紫外光谱（UV）、红外光谱（IR）、质谱（MS）和热重分析（TGA）对BMC的组成和结构进行了表征。采用MTT法分析BMC对SMMC - 7721细胞增殖的影响。采用PI染色法和Annexin - V/FITC双染色法分析BMC对SMMC - 7721细胞周期和凋亡的影响。采用荧光定量RT - PCR法分析BMC对Bcl - 2基因和Bax mRNA表达的影响，采用流式细胞术分析BMC对Bcl - 2蛋白和Bax蛋白表达的影响。全面探究了BMC的抗肿瘤活性及作用机制。结果表明，成功制备了三种新型的BMC（摩尔比为2∶1），配合物分子式分别为：Na2Ni(C21H16O11)2·10H2O、Na2Co(C21H16O11)2·8H2O和Na2Cu(C21H16O11)2·8H2O。根据细胞周期和凋亡检测结果，BMC使细胞阻滞于G0/G1期到S期以及G2/M期。基因和蛋白检测表明，在给定浓度和时间下，BMC可下调SMMC - 7721细胞中Bcl - 2基因的表达，并显著降低Bcl - 2蛋白的表达，同时，随着Bax基因表达的增加，Bax蛋白的表达也显著增加。这表明BMC通过阻滞细胞周期、降低Bcl - 2表达和增加Bax表达来抑制细胞增殖和诱导细胞凋亡；三种配合物的抗肿瘤活性为：黄芩苷 - 铜（BC - Cu）＞黄芩苷 - 钴（BC - Co）＞黄芩苷 - 镍（BC - Ni）＞黄芩苷（BC），呈现剂量 - 效应关系。
