小檗碱对结肠癌细胞HT-29增殖、凋亡及细胞膜钾通道的作用

Effects of berberine on the proliferation and apoptosis of colonic carcinoma cell line HT-29 and its mecbanism

目的观察小檗碱（Berberine，以下简称“Ber”）对结肠癌细胞株HT-29增殖、凋亡的作用并探讨其离子通道机制。方法Ber0．1—30．0μmol／L处理HT-29细胞。氮蓝四唑盐法检测细胞增殖，流式细胞术检测凋亡率，膜片钳检测延迟整流钾通道[IK（V）]。结果0．1～30．0μmol／L Ber均可抑制HT-29细胞增殖（P〈0．05），其抑癌率与作用时间及浓度相关；HT-29细胞经Ber处理24h后，凋亡率明显增高（P〈0．05），0．1、0．3、3．0、30．0μmol／LBer组凋亡率分别为（7．6±1．8）％、（9．8±2．1）％、（22．3±4．2）％、（40．6±4．5）％，对照组凋亡率为（2．4±1．1）％；0．3、3．0、30．0μmol／L的Ber能浓度依赖性抑制结肠癌细胞IK（V）（P〈0．01），阶跃刺激为＋80mV时，对照组、0．3、3．0、30．0μmol／LBer组IK（V）分别为（488±42）、（376±22）、（296±25）、（225±34）pA，0．3、3．0、30．0μmol／LBer处理组IK（V）分别为对照组的（77．16±5．41）％、（61．35±6．09）％、（45．87±7．62）％。结论Ber具有抑制结肠癌细胞HT-29增殖并诱导其凋亡的作用，此作用可能与Ber抑制肿瘤细胞细胞膜钾离子通道开放有关。

Objective To investigate the effect of berberine on the proliferation and apoptosis of colonic carcinoma cell line HT-29 and clarify its ion mechanism. Methods After treatment with Berberine （0.1-30.0 μmol/L）,cell proliferation was assayed by MTT method, cell apoptosis measured by flow cytometry,and IK（V） detected by using patch clamp technique. Results 0. 1-30.0 μmoL/L berberine could inhibit the proliferation of HT-29 cell line significantly （ P 〈 0.05 ）. After treatment with 0.1-30.0 μmol/L berberine for 24 h, the number of apoptosis cells was increased dose-dependently from （2.4 ± 1.1）% in PSS group to （7.6 ±1.8）% ,（9.8 ±2.1）% ,（22.3 ±4.2%） ,（40.6 ±4.5）% in berberine group, respectively. 0.3,3.0, and 30.0 μmol/L berberine could inhibit IK（V） of HT-29 cells significantly （ P 〈 0. 01 ）. When depolarizing at ＋ 80 mV, the IK （V） of PSS group and 0. 3,3. 0, 30.0 μmoL/L berberine-treated groups was （488 ± 42 ）, （ 376 ± 22 ）, （296 ± 25 ）, and （ 225± 34） pA respectively. The IK （V） in 0.3-30.0 μmol/L berberine-treated groups was （ 77.16 ±5.41 ） %, （ 61.35 ±6.09 ） %, and （45.87 ±7.62） % respectively compared with PSS group. Conclusion Berberine can not only inhibit the proliferation of colonic carcinoma cell line HT-29, but also induce its apoptosis, which may be due to the inhibitory effect of berberine on the delayed-rectifier potassium channel of colonic carcinoma cells.