Optimization of the preparation process of Chuanxiong extract and study on its immunomodulatory effect
Objective Optimize the preparation process of Chuanxiong extract and to study the safety and immune activity of Chuanxiong extract on immunosuppressed mice.Methods Firstly,the L9(34)orthogonal experimental method was used to optimize the extraction process of Ligusticum Chuanxiong by examining three factors:extraction time,extraction frequency,and water addition,with the transfer rate of ferulic acid and Ligusticum Chuanxiong polysaccharides as the evaluation indicators.Furthermore,the extract of Ligusticum Chuanxiong was concentrated and divided into two parts.One part was concentrated,the other part was concentrated and then alcohol precipitated.Merge concentrated solution and alcohol precipitate,and the ratio of Ligusticum Chuanxiong concentrate and alcohol precipitate was compared and determined based on appearance and taste as indicators to prepare the extract of Ligusticum Chuanxiong.Subsequently,the mice immunosuppressive model was prepared by intraperitoneal injection of cyclophosphamide.The low,medium,and high doses of Chuanxiong extract in mice were administered by gavage at 190.0,380.0 and 570.0 mg/kg,respectively.After continuous gavage for 32 days,the mouse organ index was measured,and the ELISA method for detecting mouse immunoglobulins(IgG,IgM),complement C3,C4,interleukin-6(IL-6),interleukin-8(IL-8),and tumor necrosis factor(TNF-α)Horizontal changes,MTT method to detect the transformation rate of splenic lymphocytes,and LDH method to measure NK cell activity.Results The optimal extraction process parameters for ferulic acid and Chuanxiong polysaccharides are twice,with a liquid to material ratio of 10∶1 for the first extraction and an extraction time of 4 hours;the second extraction had a liquid to material ratio of 5∶1 and an extraction time of 2 hours.When the ratio of Chuanxiong water extract concentrate to the lower layer of 65％water extract and alcohol precipitation is 3∶2,the prepared Chuanxiong extract solution has high clarity and good taste.Compared with the model group,the thymus index and spleen index were significantly increased(P＜0.01,P＜0.001),and the IgG,IgM,C3,and C4 contents in each dose group of Chuanxiong extract were significantly increased(P＜0.001,P＜0.01,P＜0.05).The high-dose group of Chuanxiong extract had IL-6,IL-8,and TNF-α levels.The content was significantly reduced(P＜0.001,P＜0.01,P＜0.05),The content of IL-6 and IL-8 in the medium dose group of Chuanxiong was significantly reduced(P＜0.001),while the content in the low dose group of Chuanxiong was significantly reduced(P＜0.01),the levels of NK cell activity and spleen lymphocyte transformation rate in immunocompromised animals in all the three dose groups of Chuanxiong extract were significantly increased(P＜0.001,P＜0.01,P＜0.05).Conclusion The optimized preparation process of Chuanxiong extract is feasible,simple,stable,with high clarity,good taste and high security;It has an immunomodulatory effect on immunosuppressive mice induced by cyclophosphamide.
Chuanxiongorthogonal test methodimmune regulationcell factorCyclo-phosphamide
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