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目的探讨苦参碱(Ma)对人横纹肌肉瘤(RMS)细胞RD增殖及凋亡的影响。方法取对数生长期的细胞,分为对照组(无Ma);Ma组:Ma终浓度分别为0.5、1.0、1.5、2.0 mg/mL;U0126(MAPK抑制剂)组:U0126为10μmol/L;Ma联合U0126共同作用组:U0126均为10μmol/L,Ma浓度同Ma组;每组设5个复孔。给予相应药物作用后,采用倒置显微镜观察细胞形态学变化;四甲基氮唑蓝(MTT)比色法检测RD细胞的增殖抑制率;流式细胞仪(FCM)检测凋亡率。结果药物作用后显微镜下可见细胞数目减少,细胞变小、变圆,随着Ma作用浓度的增加,RD细胞凋亡细胞逐步增多;Ma与U0126联合作用后,凋亡细胞明显增多,MTT比色法检测其抑制率按分组顺序分别为(13.39±1.13)%、(23.49±0.39)%、(38.84±0.35)%、(48.05±0.82)%、(63.47±1.35)%、(13.91±0.63)%、(24.52±0.46)%、(39.82±0.58)%、(48.98±0.31)%和(65.09±0.85)%;流式细胞仪检测其凋亡率按分组顺序分别为(5.49±0.96)%、(14.23±0.70)%、(25.84±4.17)%、(36.08±3.68)%、(42.53±2.71)%、(6.72±1.52)%、(17.23±3.42)%、(27.83±1.29)%、(38.25±1.44)%和(47.79±1.35)%;除U0126组外,其余各组与对照组比较差异均有显著性(P<0.05)。结论苦参碱单独或与U0126共同作用RD细胞,可抑制该细胞的增殖,促进其凋亡,在本实验范围内浓度越高这种抑制增殖、诱导凋亡的作用越明显。
Objective To investigate the effects of matrine on RD proliferation and apoptosis of human rhabdomyosarcoma (RMS) cells. Methods Logarithmic growth phase cells were divided into control group (no Ma); Ma group: Ma final concentration was 0.5, 1.0, 1.5, 2.0 mg/mL; U0126 (MAPK inhibitor) group: U0126 was 10 μmol/L. Ma combined U0126 co-action group: U0126 are 10μmol/L, Ma concentration is same as Ma group; each group has 5 complex holes. After administration of the corresponding drugs, the morphology of the cells was observed using an inverted microscope; the proliferation inhibition rate of RD cells was detected by MTT colorimetric method; and the apoptosis rate was measured by flow cytometry (FCM). RESULTS: After exposure to the drug, the number of cells decreased, and the cells became smaller and rounder under the microscope. As the concentration of Ma increased, the apoptotic cells of RD cells gradually increased; after the combination of Ma and U0126, the number of apoptotic cells increased significantly. The inhibitory rate was detected in the order of (13.39±1.13)%, (23.49±0.39)%, (38.84±0.35)%, (48.05±0.82)%, (63.47±1.35)%, (13.91±0.63), respectively. %, (24.52±0.46)%, (39.82±0.58)%, (48.98±0.31)%, and (65.09±0.85)%; flow cytometry detected the apoptotic rate in groups of (5.49±0.96)% (14.23±0.70)%, (25.84±4.17)%, (36.08±3.68)%, (42.53±2.71)%, (6.72±1.52)%, (17.23±3.42)%, (27.83±1.29)%, (38.25±1.44)% and (47.79±1.35)%; Except for U0126 group, there was significant difference between the other groups and the control group (P<0.05). Conclusions Matrine alone or in combination with U0126 can inhibit the proliferation of RD cells and promote its apoptosis. The higher the concentration of matrine in this experiment, the more obvious the effect of inhibiting proliferation and inducing apoptosis.