陈皮、枫香脂、没药、木香挥发油对人高转移卵巢癌细胞HO-8910PM体外增殖影响的研究

Effects of volatile oils extracted from Pericarpium Citri Reticulatae,Resina Liquidambaris,Myrrha and Radix Aucklandiae on the proliferation of HO-8910PM cancer cells in vitro

目的体外培养法研究陈皮、枫香脂、没药、木香挥发油对人高转移卵巢癌细胞HO-8910PM增殖的影响。方法将体外培养的HO-8910PM细胞分为挥发油组、阳性对照组、阴性对照组及Control组,分别以挥发油、环磷酰胺、DMSO及完全培养基处理后,以MTT法评价不同浓度及不同作用时间下陈皮、枫香脂、没药、木香挥发油对HO-8910PM细胞的生长抑制作用;以Hoechst33258荧光染色法染色细胞核,倒置显微镜观察细胞核形态变化;测定细胞培养液乳酸脱氢酶(LDH)、Caspase-3酶活性,评价4种挥发油对细胞膜结构及细胞凋亡的影响。结果陈皮、枫香脂、没药、木香挥发油对HO-8910PM细胞增殖具有抑制作用,并且呈时间剂量依赖性。Hoechst33258荧光染色法观察细胞核有明显的凋亡形态变化。各实验浓度挥发油处理后,细胞培养液中LDH活性、细胞中Caspase-3酶活性均明显增加,较Control组及阳性药环磷酰胺组差异有统计学意义(P<0.01)。结论陈皮、枫香脂、没药、木香挥发油对HO-8910PM细胞具有增殖抑制及促凋亡作用,其机制可能为破坏肿瘤细胞的细胞膜,激活细胞凋亡途径中的关键酶Caspase-3。

Objective To study the effects of volatile oils extracted from Pericarpium Citri Reticulatae,Resina Liquidambaris,Myrrha and Radix Aucklandiae on the proliferation of human high metastatic ovarian cancer cell line HO-8910 PM in vitro. Methods The HO-8910 PM cells cultured in vitro were divided into the volatile oils group,positive control group,negative control group and control group,which treated with volatile oils,cyclophosphamide,DMSO and complete medium respectively. The inhibition effects of volatile oils of Pericarpium Citri Reticulatae, Resina Liquidambaris, Myrrha and Radix Aucklandiae with different concentrations and different time on HO-8910 PM cells growth were evaluated by methyl thiazolyl tetrazolium( MTT) assay. The nuclei were stained by fluorescent staining Hoechst33258 and the morphological changes were observed by inverted microscope. The activity of lactate dehydrogenase( LDH) and Caspase-3 in cell culture medium were determined to evaluate the effects of four kinds of volatile oils on cell membrane structure and cell apoptosis. Results All above volatile oils exhibited inhibitive effect on the proliferation of HO-8910 PM cells in a time-dose dependent manner. The obvious morphological changes of apoptosis in nuclei were observed by Hoechst33258 fluorescence staining. After the treatment with each volatile oil at experimental concentration,the activity of LDH and Caspase-3 in cell culture medium were obviously increased,the differences were statistically significant compared with the control group and positive control group of cyclophosphamide( P < 0. 01). Conclusion The volatile oils of Pericarpium Citri Reticulatae,Resina Liquidambaris,Myrrha and Radix Aucklandiae show the effects of proliferation inhibition and apoptosis promotion on HO-8910 PM cells. The mechanism may be to destroy the cell membrane of tumor cells and activate Caspase-3 as the key enzyme in the pathway of apoptosis.