余庆花椒精油挥发性成分分析及其对红白血病HEL细胞分化和凋亡的影响

Volatile component analysis of Yuqing Zanthoxylum bungeanum essential oil and its effect on differentiation and apoptosis of red leukemia HEL cells

中文关键词:  余庆花椒精油  挥发性化合物  HEL细胞  细胞凋亡

英文关键词:Yuqing Zanthoxylum bungeanum essential oil  volatile component  HEL cells  cell differentiation  apoptosis

中文摘要:

      目的 研究余庆花椒精油(Yuqing Zanthoxylum bungeanum essential oil, YZBO)的挥发性成分及YZBO对红白血病HEL细胞分化和凋亡的影响。方法 采用顶空固相微萃取-气相色谱-质谱法(headspace solid-phase microextraction-gas chromatography-mass spectrometry, HS-SPME-GC-MS)分析YZBO的挥发性成分, 并以不同质量浓度YZBO处理HEL细胞24 h后, 采用MTT和台酚蓝计数分析细胞存活, 流式细胞仪检测HEL细胞CD71、CD235a、CD41的分化情况, 流式细胞仪和Hoechst33258染色法检测HEL细胞凋亡情况。结果 YZBO含有99种化学成分, 包括28种醇类物质、7种酯类物质、1种醛类物质、6种酸类物质、4种酮类物质、37种烯烃类、7种烷烃类物质、2种炔烃类物质、2种酚类物质和5种其他物质。橙花叔醇、榄香醇和τ-杜松醇是主要的醇类物质, 乙酸苯乙酯和异乔木萜醇乙酸酯是其主要酯类物质。YZBO对HEL细胞的半抑制浓度(median inhibitory concentration, IC50)值为2.94386 μg/mL, 可促进CD71 (45.6%)和CD235a (38.8%)的分化, 而对CD41的分化不明显, 可诱导HEL细胞程序性凋亡, 凋亡数目呈剂量和时间依赖性, 镜下和染色观察发现细胞体积变小, 核固缩以及呈高亮蓝色等典型细胞凋亡形态学改变。结论 YZBO的主要挥发性成分是烷烃类、醇类、烯烃类化合物, YZBO具有抗红白血病HEL细胞的生物活性, 可促进分化未成熟的HEL细胞向成熟细胞方向分化, 抑制向巨核细胞方向分化, 并诱导其凋亡。

英文摘要:

      Objective To study the volatile components of Yuqing Zanthoxylum bungeanum essential oil (YZBO) and the effects of YZBO on differentiation and apoptosis of erythroleukemia HEL cells. Methods The volatile component of YZBO were analyzed by headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). HEL cells were treated with different concentrations of YZBO for 24 h, the cell survival was analyzed by MTT and trypan blue counting, the differentiation of CD71, CD235a, CD41 of HEL cells were detected by flow cytometry, and the apoptosis of HEL cells was detected by Hoechst33258 staining. Results The YZBO contained 99 kinds of material composition, including 28 kinds of alcohols, 7 kinds of esters, 1 kind of aldehyde, 6 kinds of acids, 4 kinds of ketones, 37 kinds of alkenes, 7 kinds of alkanes, 2 kinds of alkynes, 2 kinds of phenols and 5 kinds of other substances. Nerolidol, elemol and τ-cadinol were the main alcohols, while phenethyl acetate and isoarborinol acetate were the main esters. The median inhibitory concentration (IC50) value of YZBO on HEL cells was 2.94386 μg/mL, which could promote the differentiation of CD71 (45.6%) and CD235a (38.8%) and had statistical significance, but did not have statistical significance on the differentiation of CD41. It could induce programmed apoptosis of HEL cells, and the number of apoptosis was dose and time dependent. Microscopic and staining observations showed that the cell volume became smaller, nuclear pyknosis and bright blue and other typical morphological changes of apoptosis. Conclusion The main volatile compounds of YZBO are mainly alkanes, alcohols and alkenes, YZBO has the biological activity of anti-erythroleukemia HEL cells, which can promote the differentiation of HEL cells to mature cells, inhibit the differentiation to megakaryocytes, and induce their apoptosis.