复方铁线蕨颗粒中总黄酮和总酚的提取及其对宫颈癌细胞的作用

Extraction of total flavonoids and total phenol in compound Adiantum capillus-veneris granules and its effect on cervical cancer cells

中文关键词:  宫颈癌  总黄酮提取物  总酚提取物  细胞增殖  细胞凋亡

英文关键词:cervical cancer  total flavonoid extract  total phenolic extract  cell proliferation  apoptosis

中文摘要:

      目的 提取复方铁线蕨颗粒中的总黄酮及总酚研究其对SiHa、HeLa、C-33A等宫颈癌细胞的增殖和凋亡的作用。方法 从复方颗铁线蕨粒中按提取工艺提取并纯化得到总黄酮及多酚提取物, 制作芦丁及没食子酸的标准曲线并采用紫外分光光度法测定含量; 将不同浓度的总黄酮及总酚给予3种细胞, 通过药物细胞毒性实验测定各细胞在2种不同提取物作用下的IC50值。以1/2IC50、IC50、2IC50作为低、中、高浓度给药, 采用异硫氰酸荧光素(fluorescein isothiocyanate, FITC)偶联Annexin-V染色法检测细胞凋亡。结果 复方铁线蕨颗粒剂中总酚含量为30.83%, 总黄酮含量39.14%。与正常对照组相比, 不同浓度的2种提取物给药下的SiHa、HeLa和C-33A细胞, 随着浓度的增加抑制率增加。在总酚作用下SiHa细胞IC50为367 μg/mL, HeLa细胞IC50为380 μg/mL, C-33A细胞的IC50为256 μg/mL。在总黄酮作用下SiHa细胞IC50为268 μg/mL, HeLa细胞IC50为274 μg/mL, C-33A细胞的IC50为110 μg/mL。在总酚及总黄酮高中低浓度作用下3种细胞的凋亡率与正常对照组相比具有显著差异, 且各个细胞总酚给药组和总黄酮给药组组内差异显著。结论 复方铁线蕨颗粒剂提取的总酚及总黄酮能抑制SiHa、HeLa、C-33A细胞的增殖, 诱导3种宫颈癌细胞凋亡。

英文摘要:

      Objective To extract the total flavonoids and total phenol in compound Adiantum capillus-veneris granules and study its effect on cervical cancer cells such as SiHa, HeLa and C-33A. Methods Total flavonoids and polyphenol extracts were extracted and purified from the compound ferrogonium fern by extraction process. The standard curves of rutin and gallic acid were prepared and determined by ultraviolet spectrophotometry. The total flavonoids and total phenols of different concentrations were given to 3 kinds of cells, and the IC50 values of each cell under the action of 3 different extracts were determined by a drug cytotoxicity test. 1/2 IC50, IC50, and 2 IC50 were administered as low, medium, and high concentrations, and apoptosis was detected by FITC-conjugated Annexin-V staining. Results The total phenolic content of compound A. fern granules was 30.83%, and the total flavonoid content was 39.14%. The SiHa, HeLa and C-33A cells administered with different concentrations of the 2 extracts increased the inhibition rate with the increase of the concentration compared with the normal control group. Under the action total phenol, the IC50 of SiHa cells was 367 μg/mL, the IC50 of HeLa cells was 380 μg/mL, and the IC50 of C-33A cells was 256 μg/mL. Under the action of total flavonoids, the IC50 of SiHa cells was 268 μg/mL, the IC50 of HeLa cells was 274 μg/ mL, and the IC50 of C-33A cells was 110 μg/mL. The apoptotic rate of the 3 kinds of cells under the action of total phenol and total flavonoids was significantly different from that of the normal control group, and the difference between the total phenol-administered group and the total flavonoid-administered group was significant. Conclusion Total phenol and total flavonoids extracted from compound Adiantum capillus-veneriscan can inhibit the proliferation of SiHa, HeLa and C-33A cells and induce apoptosis of 3 kinds of cervical cancer cells.