丁香油超高压提取工艺的优化及其抗肿瘤研究

Optimization of the extraction process of clove oil from clove buds and its antitumor activity

目的优化丁香油的提取工艺并研究其抗肿瘤活性。方法采用超高压提取丁香干燥花蕾中的精油,通过单因素试验与正交试验优化其取工艺;并通过CCK-8法检测丁香油对正常结肠上皮细胞、HT29和SW480两种结肠癌细胞的增殖抑制情况;采用Western Blot法检测丁香油处理后的两种结直肠癌细胞中凋亡相关蛋白的表达情况。结果通过单因素与正交试验确定的最佳提取条件为:粉碎粒度60目,料液比1:14,保压时间50 min,提取压力400 MPa;得率最高可达19.58%,相比传统水蒸气蒸馏提取法的得率提高了33.92%。CCK-8法结果显示:丁香油对HT29和SW480两种结肠癌细胞增殖的抑制呈现时间-剂量依赖性,差异具统计学意义,且对正常结肠上皮细胞无不良反应。Western Blot结果显示:与空白对照组比较,中、高浓度丁香油能显著上调细胞内促凋亡蛋白Bax、Caspase-3的表达;低、中、高浓度丁香油均能显著下调细胞内抗凋亡蛋白Bcl-2的表达。结论丁香油可抑制结肠癌细胞的生长。

OBJECTIVE To optimize the extraction process of clove oil essence,and to detect its anti-tumor activity.METHODS We extracted clove oil essence from clove buds using ultra-high pressure processing method,which was optimized by single factor experiment and orthogonal experiment.Furthermore,we performed cell counting kit-8(CCK-8)assay to assess the cytotoxicity of clove oil essence on normal colonic epithelial cell(HCoEpiC)and colorectal cancer cell lines(HT29 and SW480).Western Blot was performed to detect the expression of apoptosis-related proteins in two colorectal cancer cell lines treated with clove oil essence.RESULTS The optimum extraction conditions were determined by single factor and orthogonal experiments,which was as follows:the pressure was 400 MPa,the holding time was 50 min,the particle size was 60 meshes,and the solid/liquid ratio was 1∶14.The yield was the highest at 19.58%with a 34.47%increase compared to the hydrodistillation method.The results of CCK-8 showed that clove oil essence treatment(7.8-125μg·mL-1)was non-cytotoxic to HCoEpiC,but clove oil essence treatment significantly reduced the proliferative capability of HT29 and SW480 cells in a dose-and time-dependent way.The results also showed that clove oil essence treatment increased the apoptotic makers Bax/Bcl-2 ratio and Caspase 3 activities.CONCLUSION Clove oil essence can inhibit the growth of colorectal cancer cells.