参归补血汤提取工艺的优化及对IDA小鼠贫血的改善作用和JAK2/STAT3信号通路的影响

Optimization of the extraction process of Shengui Buxue Decoction and its effect on improving anemia and JAK2/STAT3 signaling pathway in IDA mice

目的优化当归、熟地黄、白芍、人参等4味药材(参归补血汤方)最佳水提取工艺,研究其对小鼠缺铁性贫血的改善作用。方法采用L_(9)(3^(4))正交实验设计法进行参归补血汤水提取工艺优化,采用双指标综合评分法确定最优水提工艺;通过建立小鼠缺铁性贫血模型,观察小鼠的生存状态,酶联免疫法检测血液中Hepcidin含量及缺铁性贫血相关指标;建立IL6诱导HepG2细胞模型,检测参归补血汤给药前后Hepcidin mRNA及JAK2/STAT3通路蛋白表达情况。结果以双指标综合评分法确定了参归补血汤的最佳水提取工艺为25倍量水,提取1 h,提取2次;动物实验表明,处方组小鼠生存状态明显改善,体质量明显增长,Hb、Si、TIBC及sTfr显著升高,Hepcidin及Fep显著下调;细胞实验表明,参归补血汤可显著下调HepG2细胞Hepcidin mRNA及其相关通路JAK2/STAT3蛋白的表达。结论通过双指标综合评分优化的参归补血汤提取工艺稳定可靠,适合工业化生产,为临床应用和新药的研发提供参考。参归补血汤可显著改善缺铁性贫血症状并有效调控Hepcidin及其相关通路JAK2/STAT3蛋白表达。

Objective To optimize the best water extraction process of the four crude medicines in Shengui Buxue Decoction,and to study its effect on improving iron deficiency anemia in mice.Methods The L_(9)(3^(4))orthogonal experimental design was used to optimize the water extraction process of Shengui Buxue Decoction,and the dual-index comprehensive scoring method was used to determine the optimal water extraction process.The mice survival state was observed by establishing the iron deficiency anemia model in mice.The enzyme-linked immunosorbent assay was used to detect the blood Hepcidin content and the related indexes of iron deficiency anemia.The HepG2 cell model induced by IL6 was established to detect the expression of Hepcidin mRNA and JAK2/STAT3 pathway protein before and after the administration of Shengui Buxue Decoction.Results The optimal water extraction process of Shengui Buxue Decoction was determined by the dual-index comprehensive scoring method as 25 times the amount of water,extracted for 1 hour,and extracted twice.The animal experiment showed that the survival state of the mice in the prescription group was significantly improved,and the body mass was increased significantly,Hb,Si,TIBC and sTfr were significantly increased,while Hepcidin and Fep were significantly down-regulated.The cell experiment showed that Shengui Buxue Decoction significantly down-regulated the expression of Hepcidin mRNA and its related pathway JAK2/STAT3 protein in HepG2 cells.Conclusion The extraction process of Shengui Buxue Decoction optimized by the dual-index comprehensive scoring is stable and reliable,which is suitable for industrial production and can provide a reference for clinical application and new drug research and development.Shengui Buxue Decoction can significantly improve the symptoms of iron deficiency anemia and effectively regulate the expression of Hepcidin and its related pathway JAK2/STAT3 protein.