| ObjectivesThe aim of this project to investigate the optimal extraction method of Echinacea purpurea and immunoregulatory function on normal mice and immune-suppressed mice. To provide theoretical basis for the development and clinical application of Echinacea purpurea in veterinary medicine.Methods1、Studies on content determination and extraction process of active component of Echinacea purpureaThis study was carried on the conditions of extraction process of Echinacea by ethanol refluxing method. Experimental analysis of the effect of solid-liquid ratio, extraction time, solvent concentration and temperature on chicory acid in Echinacea purpurea by orthogonal test. The content was determined by high performance liquid chromatography(HPLC), to determine the optimal extraction process of cichoric acid from Echinacea purpurea.2、Study on immunity regulation effect of Echinacea purpurea extracts on miceIn the experiment,80 mice were selected from Kunming, and the body weight was 20 ±2g, mice were randomly divided into 8 groups (n=10 in each group, half male and half female, fed separately). Groups were marked as control group, low, middle and high does test group(Echinacea purpurea extracts 0.05mg/g/d, 0.1mg/g/d,0.2mg/g/d), immune-suppr-essed model group, low, middle and high does immune-suppressed test group (Echinacea purpurea extracts 0.05mg/g/d, 0.1mg/g/d,0.2mg/g/d), test group were intraperitoneal injection of cyclophosphamide (0.1mg/g/bw),once a day for 3 days; Control group injected equal dose physiological saline with intragastric administration, Control group(ip NS 0.2mL/d). The control group and immune-suppressed model group ig 0.2mL/d saline, test groups and immune-suppressed test group were given different doses of Echinacea extract on daily, after 4 weeks, After the last administration in the fasted 12h, mice were killed. Determining the mice organ indices, phagocytosis of macrophage and serum hemolysin and partial cytokine (IL-2, IL-6, TNF-a).Result1、Studies on content determination and extraction process of active component of Echinacea purpureaThe experiments use the ethanol to extract of cichoric acid as the main active ingredients of Echinacea purpurea. The best extraction condition obtained, the ratio of solid to liquid was with 60% ethanol is 1:15, temperature of 90℃, extracted once for 1 hours. The effective content is 0.484% demarcated by HPLC. This extraction process is easy, stable and feasibility. The determination method is accurated and highly reproducible, suitable for quantitative production of Echinacea purpurea.2、Study on immunity regulation effect of Echinacea purpurea extracts on miceAs compared with the control group, Echinacea purpurea extracts high dose group, thymus index and spleen index increased significantly (P<0.01,P<0.05; low, middle and high dose test group significantly elevated the phagocytic activity (P<0.01); The serum hemolysis was increased by Echinacea purpurea extracts (P<0.05); low, middle and high dose test group IL-2 and IL-6 were not significantly improved (P>0.05), middle and high dose test group TNF-a increased significantly (P<0.01, P<0.05). As compared with the immune-suppressed model group, Echinacea purpurea extracts middle and high dose improved immune-suppressed test group significantly of the spleen index and thymus index(P<0.01), nearly recovered to the normal level; Low and middle doses immune-suppressed test group significantly improved the phagocytosis of immune-suppressed test group mouse peritoneal macrophages (P<0.01); Middle and high dose immune-suppressed test group significantly increased the serum hemolysin in immune-suppressed mice (P<0.01), up to 42.08%. Low, middle and high dose immune-suppressed test groups can obviously increase the contents of IL-2, TNF-a in immune-suppressed mice (P<0.01), The contents of IL-6 of low and high dose immune-suppressed test groups were significantly different (P<0.01), Among them, the high dose immune-suppressed group can make immune-suppressed mice IL-2, IL-6, TNF-levels were increased by 38.49%,41.63%,67.38%.ConclusionModified the extraction process of Echinacea by ethanol refluxing method, the extraction process is simple, and easy to operate, and the content of the active ingredients in the extraction is high. Echinacea purpurea extracts can improve the immune function of normal mice and immune suppression mice, In contrast, the regulation of immune function in mice with immune suppression is better. |
