血复康组方对HEL细胞增殖抑制和诱导分化的研究
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摘要
[目的]
通过体外研究血复康主要组分青黛、葛根、莪术的有效抗肿瘤成分,观察其对具有JAK2突变的人红白血病HEL细胞株增殖抑制、诱导分化的影响。
[方法]
分别提取青黛、莪术、葛根中的有效抗肿瘤成分,使其有效成分中靛玉红、莪术油及葛根总黄酮纯度>80%,配制所需浓度的溶液。选择不同浓度的青黛提取物、莪术提取物、葛根提取物作用于HEL细胞,MTT法检测药物作用24h、48h对细胞增殖抑制的影响；应用流式细胞术检测诱导细胞分化过程中细胞表面抗原CDllb、CD14的表达情况；RT-PCR在]mRNA水平检测细胞分化标志GPA、GATA1基因表达变化。
[结果]
1.选择青黛提取物浓度25-100ug/ml、莪术提取物浓度10-80ug/ml及葛根提取物浓度50-400ug/ml作用于HEL细胞。青黛、莪术提取物均能抑制HEL细胞增殖,且呈浓度-时间依赖性效应关系；葛根提取物低浓度(≤200ug/ml)促进细胞增殖,高浓度(>200ug/ml)抑制细胞增殖。
2.青黛提取物、莪术提取物能在一定浓度范围内能增加细胞表面CDllb、CD14等表达,青黛50ug/ml、莪术20ug/ml作用显著；葛根提取物无明显作用。
3.RT-PCR显示：青黛提取物50ug/ml、莪术提取物20ug/ml均能上调HEL细胞分化分子标志。GPA、GATA1的mRNA表达。
[结论]
血复康组方青黛提取物、莪术提取物及高浓度葛根提取物对人红白血病HEL细胞有增殖抑制作用；青黛、莪术提取物能有效诱导HEL细胞分化。
【Objective】To observe the effects of the active ingredients of XueFuKang(XFK) on leukemic cell-HEL proliferation inhibition and differentiation.
[Method] HEL cells were treated with the extractions of indigo, Kudzuvine Root, Zedoray Rhizome, the effective ingredients of XFK, respectively, at varied concentration for 24 and 48h. MTT assays were used to measure the inhibitory effects of cell proliferation. The differentiation of HEL cells was revealed by flow cytometry marked with CD11b、CD14, while the expression of GPA, GATA1 was detected via RT-PCR.
【Results】
1. The proliferation of HEL cells was inhibited by the extraction of Indigo at 25-100ug/ml, Zedoray Rhizome at 10-80ug/ml respectively in a concentration and time dependent manner,and was promoted in low concentrations(<200 ug/ml),while inhibited in high concentrations(>200 ug/ml) by that of Kudzuvine Root.
2. The expression of CD11b,CD14 was increased in a certain range of concentration with the extraction of indigo and Zedoray Rhizome.and maximal expression level of the two antigens was observed at 50ug/ml for the extraction of Indigo,20ug/ml for that of Zedoray Rhizome respectively. But this phenomenon was not observed in that of Kudzuvine Root.
3. The mRNA level of GPA, GATA1, the cell differentiation markers of HEL cells, was upregulated at 50ug/ml for Indigo,20ug/ml for Zedoray Rhizome using RT-PCR.
[Conclusion] There is an inhibitory effect on the proliferation of HEL cells treated with all of three individual extracts from XFK, the extractions of indigo and Zedoray Rhizome can promote the HEL cells differentiation,which is conceived by the CD11b,CD14 antigens expression on cell surface and GPA, GATA1 mRNA expressions in HEL cells.
通过体外研究血复康主要组分青黛、葛根、莪术的有效抗肿瘤成分,观察其对具有JAK2突变的人红白血病HEL细胞株增殖抑制、诱导分化的影响。
[方法]
分别提取青黛、莪术、葛根中的有效抗肿瘤成分,使其有效成分中靛玉红、莪术油及葛根总黄酮纯度>80%,配制所需浓度的溶液。选择不同浓度的青黛提取物、莪术提取物、葛根提取物作用于HEL细胞,MTT法检测药物作用24h、48h对细胞增殖抑制的影响；应用流式细胞术检测诱导细胞分化过程中细胞表面抗原CDllb、CD14的表达情况；RT-PCR在]mRNA水平检测细胞分化标志GPA、GATA1基因表达变化。
[结果]
1.选择青黛提取物浓度25-100ug/ml、莪术提取物浓度10-80ug/ml及葛根提取物浓度50-400ug/ml作用于HEL细胞。青黛、莪术提取物均能抑制HEL细胞增殖,且呈浓度-时间依赖性效应关系；葛根提取物低浓度(≤200ug/ml)促进细胞增殖,高浓度(>200ug/ml)抑制细胞增殖。
2.青黛提取物、莪术提取物能在一定浓度范围内能增加细胞表面CDllb、CD14等表达,青黛50ug/ml、莪术20ug/ml作用显著；葛根提取物无明显作用。
3.RT-PCR显示：青黛提取物50ug/ml、莪术提取物20ug/ml均能上调HEL细胞分化分子标志。GPA、GATA1的mRNA表达。
[结论]
血复康组方青黛提取物、莪术提取物及高浓度葛根提取物对人红白血病HEL细胞有增殖抑制作用；青黛、莪术提取物能有效诱导HEL细胞分化。
【Objective】To observe the effects of the active ingredients of XueFuKang(XFK) on leukemic cell-HEL proliferation inhibition and differentiation.
[Method] HEL cells were treated with the extractions of indigo, Kudzuvine Root, Zedoray Rhizome, the effective ingredients of XFK, respectively, at varied concentration for 24 and 48h. MTT assays were used to measure the inhibitory effects of cell proliferation. The differentiation of HEL cells was revealed by flow cytometry marked with CD11b、CD14, while the expression of GPA, GATA1 was detected via RT-PCR.
【Results】
1. The proliferation of HEL cells was inhibited by the extraction of Indigo at 25-100ug/ml, Zedoray Rhizome at 10-80ug/ml respectively in a concentration and time dependent manner,and was promoted in low concentrations(<200 ug/ml),while inhibited in high concentrations(>200 ug/ml) by that of Kudzuvine Root.
2. The expression of CD11b,CD14 was increased in a certain range of concentration with the extraction of indigo and Zedoray Rhizome.and maximal expression level of the two antigens was observed at 50ug/ml for the extraction of Indigo,20ug/ml for that of Zedoray Rhizome respectively. But this phenomenon was not observed in that of Kudzuvine Root.
3. The mRNA level of GPA, GATA1, the cell differentiation markers of HEL cells, was upregulated at 50ug/ml for Indigo,20ug/ml for Zedoray Rhizome using RT-PCR.
[Conclusion] There is an inhibitory effect on the proliferation of HEL cells treated with all of three individual extracts from XFK, the extractions of indigo and Zedoray Rhizome can promote the HEL cells differentiation,which is conceived by the CD11b,CD14 antigens expression on cell surface and GPA, GATA1 mRNA expressions in HEL cells.
引文
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[6]戴海萍,周建伟,朱光荣.青黛复方对K562细胞bcr/abl及JWA基因表达的影响[J].中国实验血液学杂志,2005,(05)：809.
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