| This experiment was designed to study extracting conditions of Pumpkin (Cucurbita moschata) leaves by ethanol and the main chemical composition of the ethanol extract, and also focus on acute toxicity and immunomodulatory of Pumpkin (Cucurbita moschata) leaf ethanol extract. The main conclusions are as follows:1. By determining polyphenol compounds and vitamin C content, drying Pumpkin (Cucurbita moschata) leaves at50℃was selected after compared fresh leaves and dried leaf powders at50℃and100℃, respectively. Furthermore, content of the polyphenol compounds and vitamin C in water, acetone, methanol, ethyl acetate extracts of Pumpkin (Cucurbita moschata) leaves were compared. A comprehensive analysis suggested that the selection of ethanol be the best for extracting Pumpkin (Cucurbita moschata) leaves.The result of single factor test and orthogonal test that are based on polyphenol compounds and vitamin C content index in Pumpkin (Cucurbita moschata) leaf ethanol extract showed that the effective order was as following:ethanol concentration> temperature> time> solid-liquid ratio, and the optimum extraction conditions were55%ethanol, extraction temperature50℃, extraction time2h and solid-liquid ratio1:16g/mL. At the same time, content of main components in the extract, including water, crude protein, crude fat, total sugar, ash,was10.32±0.58%,15.96±0.71%,5.01±0.12%,29.62±0.69%,10.77±0.11, respectively. Furthermore, polyphenol compounds and vitamin C content in the extract reached to35.24±0.37mg/g and8.73±0.47mg/g, respectively.2.The result of acute toxicity test showed that ethanol extract of Pumpkin (Cucurbita moschata) leaves are non-toxic. No abnormal animals were observed in each dose group. LD50of ethanol extract of Pumpkin(Cucurbita moschata) leaves was more than18.0g/kg· bw. According to classification standard, the ethanol extract is non-toxic. Furthermore, the extract did not have any significant effect on the growth performance of mice, and there was no correlation between gender and weight gain or food utilization of mice.It was also found that hematological indexes of each group were within the normal range, except for the slightly lower white blood cell count of female in18.0g/kg· bw group and male in12.0g/kg· bw and18.0g/kg· bw groups. Furthermore, white blood cells and platelets indexes showed more significant difference compared with other group at higher doses (P＜0.05) while different gender also showed significant difference (P<0.05). The dose below8.0g/kg· bw did not show significant difference between all testing groups. Although the thymus index showed a downward trend, and spleen index actually showed a upward trend with increase in ethanol extract dosage, all organ coefficients were within normal range.3. The result indicated that growth trend of weight in each group were similar before injection of cyclophosphamide, which did not show significant difference between each other (P>0.05). After injection of cyclophosphamide, the weight of mice in injected group lost, while the normal group did not change significantly (P>0.05). This result indicated that artificial immunocompromised was successful. Meanwhile, the organs of mice in each group did not show any lesions, and their color and shape were normal. Organ coefficients were within normal range. Particularly, it was proved that the ethanol extract had the hepatoprotective effect on liver and promoting effect on the growth of immune organs.Immunomodulatory effects studies have shown that Pumpkin leaf ethanol extract could promote the immune function of mice. The extract could promote T-lymphocyte proliferation and T cell-mediated immune response. It also could promote the ability of antibody formation in mice at higher doses. The experimental result did not show any significant impact on secretion of serum hemolysin. It was also showed that Pumpkin leaf ethanol extract, which had no significant effect with normal mice, can strengthen the ability of peritoneal macrophage in mice to absorb chicken erythrocytes and promote carbon clearance phagocytic capacity of enhanced immunocompromised mice phagocytes in appropriate doses. |
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