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地黄活血汤调控IL-6表达治疗子宫平滑肌瘤活性成分的筛选和测定
曾秀芳,谭梅英
0
(佛山市南海区第九人民医院,广东 佛山,528203;广东省第二中医院(广东省中医药工程技术研究院),广东 广州,510095)
摘要:
目的:筛选并准确测定地黄活血汤调控白细胞介素(IL)-6表达治疗子宫平滑肌瘤的活性成分,作为其基于功效的质量评价指标。方法:活性筛选采用酶联免疫吸附(ELISA)法,分别加入地黄活血汤中所含各成分单体,测定其对IL-6表达的影响,将产生显著影响者列入候选;以上述候选成分为指标,进行高效液相色谱法(HPLC)含量测定研究,使用Waters Xbridge C18色谱柱(4.6 mm×250 mm,5 μm),乙腈-0.2%磷酸为流动相,梯度洗脱,流速1.0 ml/min,检测波长254 nm,柱温30℃,进样量10 μl;选择毛蕊花糖苷、木通皂苷D为指标性成分,共测定10批药材制备的供试品溶液。结果:ELISA法检测出毛蕊花糖苷、木通皂苷D、川续断皂苷乙、表儿茶素、芍药苷具有较为显著的IL-6表达调控活性;含量测定结果表明,毛蕊花糖苷、木通皂苷D在地黄活血汤中含量分别为0.0370~0.0534 mg/ml和0.0419~0.0590 mg/ml。结论:地黄活血汤中的毛蕊花糖苷、木通皂苷D能体外抑制IL-6的表达,建议列为该方基于功效主治的质量控制指标成分。
关键词:  地黄活血汤  子宫平滑肌瘤  IL-6  毛蕊花糖苷  木通皂苷D  实验研究
DOI:
Screening and determination of the active components of Dihuang Huoxue decoction for the treatment of leiomyoma of the uterus by regulating the expression of interleukin-6
ZENG Xiufang,TAN Meiying
(The Ninth People’s Hospital of Nanhai District,Foshan 528203,Guangdong,China;Guangdong Second Provincial Traditional Chinese Medicine Hospital & Guangdong Research Institute of Traditional Chinese Medicine Manufacturing Technology,Guangzhou 510095,Guangdong,China)
Abstract:
Objective:To screen out and measure the active components of Dihuang Huoxue decoction for the treatment of leiomyoma of uterus by regulating the expression of interleukin-6 (IL-6),as function-based quality assessment indices.Methods:ELISA was used to screen out the active components,and each component of Dihuang Huoxue decoction was added to observe their influence on the expression of IL-6,among which the components with marked influence were included as candidates.To measure the content of these candidate components,high-performance liquid chromatography was performed on a Waters Xbridge C18 column (4.6 mm×250 mm,5 μm) with a mobile phase of acetonitrile-0.2% phosphoric acid for gradient elution,at a flow rate of 1.0 ml/min,a detection wavelength of 254 nm,a column temperature of 30℃,and a sample size of 10 μl.Verbascoside and akebia saponin D were selected as index components,and the test article solutions of 10 batches of medicinal material were measured.Results:ELISA showed that verbascoside,akebia saponin D,dipsacoside B,epicatechin,and paeoniflorin had a marked effect in regulating the expression of IL-6.The results of content determination showed that the content of verbascoside and akebia saponin D in Dihuang Huoxue decoction were 0.0370~0.0534 mg/mL and 0.0419~0.0590 mg/mL,respectively.Conclusion:Verbascoside and akebia saponin D in Dihuang Huoxue decoction can inhibit the expression of IL-6 in vitro and are recommended to be included as function-based quality assessment indices.
Key words:  Dihuang Huoxue decoction  leiomyoma of the uterus  interleukin-6  verbascoside  akebia saponin D  experimental study

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