三叶青水提物及其含药血清对人胃腺癌细胞株的抑制作用

Effect of Water Extract of Tetrastigmatis hemsleyani and the Serum Containing it on the Proliferation of AGS Cells

目的:考察三叶青水提物及其含药血清对人胃腺癌细胞AGS体外增殖的影响。方法:清洁级小鼠随机分为5组:三叶青水提物高、中、低(20,10,5 g.kg-1)剂量组、环磷酰胺阳性对照(CTX,5 g.kg-1)组及阴性对照组(蒸馏水)。各组均ig给药,每次20 mL.kg-1,每天2次,连续给药3 d,最后1次给药后1 h断头取血,4℃静置4 h,3 000 r.min-1离心20 min,分离血清,56℃灭活30 min后,用0.22μm的滤膜过滤除菌,-20℃保存备用。对数生长期的AGS细胞经0.25%胰蛋白酶消化后,配成2.5×104/L的细胞悬液接种于96孔培养板,培养24 h后,加入不同浓度的三叶青水提物溶液及其含药血清,作用48 h后,采用MTT比色法在570 nm波长下测定A,观察不同浓度三叶青水提物及其含药血清对AGS细胞的抑制作用。结果:1～40 g.L-1三叶青水提物对AGS细胞有显著的抑制作用(P<0.01),IC50为13.15 g.L-1;三叶青水提物含药血清高、中、低(20,10,5 g.kg-1)剂量组也可显著地抑制AGS肿瘤细胞的增殖(P<0.01),IC50为70.06 g.L-1。二者均呈现出剂量依赖性。结论:三叶青水提物及其含药血清对AGS细胞均有较强抑制作用。

Objective： To investigate the effect of the water extract of Tetrastigmatis hemsleyani and the serum containing it on the proliferation in vitro. Method： The mice of clean grade were randomly assigned into 5 groups： high dose group （20 g·kg-1） , moderate dose group （10 g ·kg-1）, low dose groups （5 g·kg-1）, positive control cyclophosphamide （CTX, 5 g ·kg-1） group and negative control group （distilled water）. Every group was administered by intragastric administration 20 mL·kg-1 each, twice a day within 3 successive days. One hour after the last administration on third day, blood was collected from every decapitated mouse. The blood was centrifugalized for 20 min at 3 000 r· min-1 after standing at 4 ℃. The separated serum was filtered to remove bacteria by 0.22 μm sepatate film at 56 ℃ after inactivated for 30 rain and stored at - 20 ℃ for later use. The suspensions of AGS cells in logarithmic growth phase digested by 0.25% tripsin were inoculated in 96 well culture plate with a cell density of 2.5 × 10^4/L. The different concentrations of the water extract of T. hemsleyani and the serum containing it were added into the culture after 24 h culture. MTT assay was used with determinating A value in the wavelength of 570 nm to evaluate the cell proliferation of the water extract of T. hemsleyani and the serum containing it for 48 h. Result： When the concentration of the water extract of T. hemsleyani was range from 1 g · L-1 to 40 g ·L-1 , a significant proliferation inhibiting （P 〈0.01 ） was shown and its 50% inhibition concentration （ICs0） was 13.15 g ·L-1. High, moderate and low dose treatment groups （20, 10, 5 g·kg-1） also significantly inhibited the proliferation of AGS cells and its ICs0was 70.06 g·L-1. Both of the proliferation inhibitings were shown in dose-dependent manner. Conclusion： The water extract of T. hemsleyani can strongly inhibit the AGS cells proliferation.