灵芝孢子粉破壁方法优化及其免疫调节和抗肿瘤机制研究

Optimize the Wall-breaking Method of Ganoderma Lucidum Spore and Research Immunomodulatory and Antitumor Mechanism

目的对灵芝孢子粉破壁优化方法及其免疫调节和抗肿瘤细胞转导的机制进行探讨。方法以灵芝孢子破壁率作为试验指标,对灵芝孢子破壁工艺进行优化。免疫调节试验方法采用剂量为150,300,600 mg·kg^(-1)BW的破壁灵芝孢子粉给小鼠连续灌胃30 d,测定并对比不同剂量的小鼠刀豆蛋白诱导下淋巴细胞转化功能、抗体生成细胞数、迟发型变态反应、溶血素水平、腹腔巨噬细胞吞噬鸡红细胞能力、单核-巨噬细胞碳廓清能力及NK细胞活性。抗肿瘤细胞试验方面,建立S180荷瘤小鼠模型,研究破壁灵芝孢子粉的抗肿瘤作用。结果破壁灵芝孢子粉在小鼠脾淋巴细胞转化与增殖方面的增强作用明显,促进抗体生成细胞增殖,提高小鼠迟发型变态反应进程,小鼠血清溶血素水平提高,腹腔巨噬细胞吞噬能力及单核-巨噬细胞碳廓清能力也得到增强;在肿瘤试验方面,人肝癌HepG2细胞在破壁灵芝孢子作用下,抑制率明显提高,达到43.4%~57.6%,肝体比温度的变化及抗肿瘤作用显著。结论经优化后的破壁工艺能显著提高灵芝孢子粉有效成分的溶出,而破壁灵芝孢子粉在增强免疫力和抗肿瘤方面均有明显的效果。

OBJECTIVE On the wall-breaking method of ganoderma lucidum spore and its immunomodulatory and antitumor cell transduction mechanism are discussed. METHODS The wall solvent, water as test factors include homogeneous, pressure and liquid ratio,number of ganoderma lucidum spore wall-broken rate as test index, optimize the ganoderma lucidum spores broken process. Immunity test method using the wall upon maturity of ganoderma lucidum spore 150,300,600 mg. kg-~ BW dose lavage for 30 days, to mice with different doses of mice under the sword bean protein induced lymphocyte transformation function, the number of antibody generating cells in mice, the mice delayed-type allergy, hemolysin level, macrophage cell in abdominal cavity chicken red blood cell capacity and mononuclear maerophage carbon clearance ability and NK cell activity were determined.The experimental aspects of tumor cells, the establishment of S180 tumor-burdened mouse model, research the antitumor function of the broken ganoderma lucidum spore. RESULTS Broken ganoderma lucidum spore in mice spleen lymphocyte transformation and proliferation of enhanced obviously, promoteed antibody generating cell proliferation, improveed mice delayed-type allergy process, mice serum hemolysin level, abdominal macrophage phagocytosis and mononuclear macrophage carbon clearance ability also enhanced; In terms of tumor experiment, human liver HepG2 cells in the wall under the action of ganoderma lucidum spores,inhibition rate increased significantly ,43.4%- 57.6%,liver body than the changes in temperature, anti-tumor effect was remarkable. CONCLUSION The break walls of the optimized process can significantly increase the dissolution of active ingredients upon maturity of ganoderma lucidum spore, and the wall in the aspect of immunity enhancement and antitumor upon maturity of ganoderma lucidum spore has the obvious effect.