广西莪术中提取姜黄素及联合顺铂诱导人卵巢癌细胞凋亡的研究

EFFECTS OFCURCUMIN EXTRACT COMBINED WITH CISPLATIN ON APOPTOSIS OF SKOV3 CELLS

目的:利用表面活性剂提高广西莪术中姜黄素提取的含量,并观察姜黄素联合顺铂对人卵巢癌SKOV3细胞凋亡的影响。方法:适量表面活性剂润湿后,醇提法提取姜黄素,用高效液相色谱法测定得到提取率。再取对数生长期的SKOV3细胞接种,分别给以25,50,100μmol/L姜黄素和联合用药组以25,50,100μmol/L姜黄素与10μmol/L顺铂联用,培养48h用MTT法测定吸光度,计算细胞生长抑制率;并设正常对照组、50μmol/L姜黄素组、10μmol/L顺铂组和50μmol/L姜黄素与10μmol/L顺铂联合用药组做细胞AO/EB染色,观察细胞凋亡情况。结果:每10g莪术原料加入10mL 1%十二烷基硫酸钠(SDS)润湿30min,80%乙醇回流提取2次,每次1.5h得到最大收益;MTT结果与荧光染色观察均表明姜黄素联合顺铂可协同诱导SKOV3细胞凋亡。结论:SDS对莪术中姜黄素的提取率有较大影响,从广西莪术中提取的姜黄素联合顺铂可协同诱导SKOV3细胞凋亡。

Objective： To improve the curcumin content extracted from Curcuma kwangsiensis with surfactant and study its effect on the apoptosis of human ovarian carcinoma SKOV3 combined with cisplatin. Methods： Moistened with appropriate quantity of surfactant, the curcumin was extracted by alcohol and assayed by HPLC for its ratio. The cells inoculated in the logarithm phase were administrated with 25, 50, and 100 umol/L of curcumin or combined medication： 25, 50, and 100 umol/L of curcumin was combined with 10 umol/L of cisplatin, respectively, cultured for 48h and their absorbances was assayed by MTT method to calculate the inhibition rate of the cell growth. The normal controls, the 50 umol/L of curcumin group, the 10 umol/L of cisplatin group, and the 50 umol/L of curcumin combined with 10 umol/L of cisplatin was AO/EB stained to observe their apoptosis. Results： A maximum yield was obtained from each 10 g of Curcuma bulk material moistened by 10 mL 1% of Sodium dodecyl sulfate （SDS） for 30 min, extracted with double refluxed ethanol for 1.5 h per time. The MTT outcome and observation of Fluorescent staining indicated a synergistic induction to SKOV3 apoptosis by curcumin and cisplatin combination. Conclusion. SDS affects significantly to the curcumin extraction from curcuma. SKOV3 apoptosis can be synergistically inducted by curcumin in Curcuma kwangsiensis and cisplatin combination.