CL联合苦参碱抑制胃癌细胞共培养内皮细胞增殖与凋亡

Effects of Rosa roxburghii Tratt extract and matrine on proliferation and apoptosis of human endothelial cells cultured with supernatant of gastric cancer cells

目的探讨刺梨提取物CL联合苦参碱(m atrine)抑制人胃癌细胞SGC-7901培养上清诱导的人脐静脉内皮细胞ECV-304的增殖和凋亡及其机制。方法人胃癌细胞SGC-7901培养上清诱导培养人脐静脉内皮细胞ECV-304,多个质量浓度苦参碱、CL单药干预,或多个质量浓度苦参碱联合CL干预,M TT法检测内皮细胞增殖抑制率,中效原理法判断联合用药的相互作用;采用RT-PCR和蛋白印迹法分析K i-67、B ax、B cl-2 mRNA/蛋白表达变化。结果10 m g/L、20 m g/L、40 m g/L、80 m g/L、160 m g/L质量浓度的CL干预48小时,其抑制率分别为(15.1±2.1)%、(23.1±2.3)%、(34.1±3.3)%、(46.9±3.8)%、(68.8±2.9)%;15 m g/L、30 m g/L、60 m g/L、120 m g/L、240 m g/L质量浓度的苦参碱干预48小时,ECV-304抑制率分别为(20.8±1.3)%、(25.1±2.2)%、(40.9±1.1)%、(62.9±2.2)%、(77.2±1.9)%;CL+苦参碱联合干预48小时,其抑制率分别为(33.8±2.1)%、(46.8±2.4)%、(84.2±2.0)%、(88.1±2.2)%、(94.8±0.9)%,以上均呈剂量依赖(P<0.05)。IC50浓度(75 m g/L)CL、IC50浓度(75 m g/L)苦参碱、1/2 IC50(CL+苦参碱)分别干预ECV-304细胞,抑制率分别为(40.7±1.3)%、(46.2±1.2)%、(51.4±0.7)%,联合用药抑制率增高(P<0.05);中效原理得出一定浓度范围两药联合为协同效应;与单独用药组比较,联合用药组K i-67、B cl-2 mRNA/蛋白表达显著降低,而B ax mRNA/蛋白表达显著增高。结论刺梨提取物CL、苦参碱均可抑制人脐静脉内皮细胞ECV-304的增殖,两药联合应用存在协同作用。

Objective To investigate the effects of Rosa roxburghii Tratt extract （CL） combined with matrine （MA） on the proliferation and apoptosis of the human endothelial cell cultured with supernatant derived from gastric cancer cells. Methods Human endothelial EVC-304 ceils cultured with supernatant of human gastric carcinoma SGC-7901 cells were exposed to CL and/or MA. MTT was used to measure cell proliferation. The expression levels of Ki-67 and Bcl-2 mRNA were detected by RT-PCR, and the expression levels of Ki-67,Bcl-2 and Bax proteins were evaluated by Western blot. Results After treated with CL （10,20,40,80 and 160 mg/L,for 48 h）,MTT assay revealed the inhibition rates were （15.1± 2.1）%, （23.1=1=2.3）%,（34.1±3.3）%,（46.9±3.8）% and （68.8±2.9）%,respectively; after treated with matrine （15,30, 60, 120 and 240 mg/L, for 48 h）, the inhibition rates were （20.8± 1.3） %, （25.1 ± 2.2） %, （40.9 ±1.1） %, （62.9 ± 2.2） and （77.2± 1.9） %, respectively ; after treated with CL combined with matrine [（10± 15）mg/L, （20 ± 30） mg/L, （40 ±60） mg/L,（80±120）mg/L and （160±240）mg/L,for 48 hi,the inhibition rates were （33.8±2.1）%,（46.8±2.4）%,（84.2± 2.0）%, （88.1±2.2）% and （94.8±0.9）%,respectively; showing a dose-dependent manner（P〈0.05）. After treated with ICsoCL（75 mg/L）,IC50 matrine（75 mg/L） and 1/2 IC50 CL combined with 1/2 IC50 matrine [（37.5±37.5）mg/L,48 h],MTT assay revealed the inhibition rates were （40.7± 1.3）%, （46.2± 1.2） % and （51.4±0.7）% ,respectively（P〈0.05） ; mediaeffect principle showed a synergistic effect of CL combined with MA. Compared with those in CL or MA alone,the levels of Ki-67 and Bcl-2 mRNA/protein expression were significantly decreased in combination group （P〈0.05）; the levels of Bax mRNA/protein expression were significantly increased in combination group（P〈0.05）. Conclusion CL/MA can inhibit the proliferation of the human endothelial cell; when CL was combined with MA,these two drugs show a synergistic effect.