三叶青水提取物对NK细胞杀伤胃癌BGC-823细胞的影响

Effect of the water extract of Tetrastigma hemsleyani on the cytotoxic activity of NK cells against gastric cancer cell lines BGC-823

目的探讨三叶青水提取物对人NK细胞杀伤胃癌BGC-823细胞的影响及机制。方法采用干细胞生长培养基（SCGM）体外扩增人外周血NK细胞,不同浓度三叶青水提取物作用于NK细胞后,采用CCK8法检测三叶青水提取物对NK细胞增殖率的影响,流式细胞术（FCM）检测NK细胞穿孔素（PFP）、颗粒酶B（Gra B）及CD107a的表达,乳酸脱氢酶（LDH）释放法检测NK细胞对胃癌BGC-823细胞的杀伤活性。结果三叶青水提取物作用人NK细胞48 h后,三叶青水提取物0.0003~78.125μg/ml浓度组的NK细胞增殖明显（P〈0.05）;三叶青水提取物0.305~4.883μg/ml浓度组人NK细胞PFP、Gra B及CD107a表达高于对照组（P〈0.05）;三叶青水提取物0.305~19.531μg/ml浓度组人NK细胞对胃癌BGC-823细胞杀伤活性较对照组增强,差异有统计学意义（P〈0.05）。结论三叶青水提取物能促进NK细胞的增殖,能增强NK细胞对胃癌BGC-823细胞的杀伤活性,这可能与三叶青水提取物上调NK细胞PFP、Gra B及CD107a表达有关。

Objective To explore the effect and mechanism of the water extract of Tetrastigma hemsleyani on the cy- totoxic activity of NK cells against gastric cancer cell lines BGC-823. Methods The amplification of human NK cells was induced by stem cell growth medium （SCGM） in vitro. NK cells with different concentrations of water extract of Tet- rastigma hemsleyani in vitro were cultured. CCK8 assay was used to measure the growth curve of NK cells. The expres- sions of perforin （PFP） , Granzyme B （GraB） and CD107a on NK cells were detected by flow cytometry （FCM）. The cytotoxic activity of NK ceils to gatric cancer BGC-823 cells was detected by lactate dehydrogenase （LDH） release. Re- suits With the water extract of Tetrastigma hemsleyani after 48 hours, the proliferation rate of human NK cells was in- creased significantly with the water extract of Tetrastigma hemsleyani at concentrations from 0. 0003 to 78. 125 μg/ml （P 〈 0.05） ; the expressions of GraB, PFP and CD107a of human NK cells with the water etract of Tetrastigma hemsley- ani at concentrations from 0. 305 to 4. 883 μg/ml were significantly higher than control group （P 〈 0.05） ; the cytotoxic activity of human NK cells against BGC-823 with the water extract of Tetrastigma hemsleyani at concentrations from 0. 305 to 19. 531 μg/ml was remarkably higher than the control group （P 〈 0.05）. Conclusion The water extract of Tetrastigma hemsleyani at a certain range of concentrations could promote the proliferation of NK ceils and enhance the cytotoxic activity of NK cells to BGC-823 cells. The possible mechanism may be that the water extract of Tetrastigma hemsleyani could increase expressions of PFP, GraB and CD107a.