Study on Immune-enhancing Activity of Gonad Saponins from Female of Apostichopus japonicus
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摘要: 目的：以仿刺参雌性生殖腺皂苷（gonad saponins from female of Apostichopus japonicus，AGS）为研究对象，探究其免疫增强活性。方法：通过对巨噬细胞吞噬能力和细胞因子的测定，研究AGS的体外免疫调节活性；通过对免疫低下小鼠胸腺、脾脏指数，碳廓清指数与吞噬指数，腹腔巨噬细胞增殖活性，脾细胞增殖活性，自然杀伤细胞（natural killer cell，NK cell）活力，T淋巴细胞亚群水平及小鼠血清细胞因子的测定，分析AGS在小鼠体内的免疫调节活性。结果：体外实验表明，AGS能提高小鼠体外巨噬细胞吞噬能力，促进TNF-α、IL-6、IFN-γ的分泌，且各组与正常组相比差异显著（P<0.05）；体内实验表明，与模型组相比，中剂量组和高剂量组AGS均能显著提高小鼠胸腺、脾脏指数、碳廓清指数和吞噬指数（P<0.05），显著增强腹腔巨噬细胞和脾细胞增殖活性（P<0.05），显著增强NK细胞的活性（P<0.05），显著提高CD4+/CD8+水平（P<0.05），显著促进TNF-α、IL-6、IFN-γ的分泌（P<0.05）。结论：AGS在体外和体内都表现出增强免疫的活性。Abstract: Objective: Taking gonad saponins from female of Apostichopus japonicus (AGS) as the research object to explore its immune-enhancing activity. Methods: The immunoregulatory activity of AGS in vitro was studied by measuring the phagocytosis and cytokines of macrophages. Thereafter the immunoregulatory activity of AGS in immunocompromised mice was analyzed by measuring thymus and spleen index, carbon clearance index and phagocytic index, proliferation activity of peritoneal macrophages, spleen cells, NK cell activity, T lymphocyte subsets level and serum cytokines of mice. Results: In vitro experiments showed that AGS could improve the phagocytic ability of macrophages and enhance the secretion of TNF-α, IL-6 and IFN-γ, and each group was significantly different from the normal group (P<0.05). In vivo experiments showed that compared with model group, AGS in middle dose group and high dose group could significantly increase thymus and spleen index, carbon clearance index and phagocytosis index (P<0.05), significantly enhance the proliferation activity of peritoneal macrophages and spleen cells (P<0.05), significantly enhance the activity of NK cells (P<0.05), significantly increase the level of CD4+/CD8+ (P<0.05), and significantly promote the secretion of TNF-α, IL-6, and IFN-γ (P<0.05). Conclusion: AGS exhibits immune-enhancing activity both in vitro and in vivo.
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Key words:
- Apostichopus japonicus /
- gonad /
- saponin /
- immune-enhancing activity
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图 2 AGS对小鼠巨噬细胞增殖活性的影响
Figure 2. Effects of AGS on proliferation of macrophages in mice
图 4 AGS对小鼠脾细胞中自然杀伤细胞（NK细胞）活力的影响
Figure 4. Effects of AGS on NK cell activity of splenocytes in mice
图 5 AGS对小鼠脾脏T淋巴细胞亚群的影响
Figure 5. Effect of AGS on T lymphocyte subsets of spleen in mice
注：A：正常组；B：模型组；C：低剂量组；D：中剂量组；E：高剂量组。
图 6 AGS对小鼠血清细胞因子的影响
Figure 6. Effects of AGS the release of cytokines in serum of mice
注：A：TNF-α；B：IL-6；C：IFN-γ；*表示与正常组相比，差异显著（P<0.05）；#表示与模型组相比，差异显著（P<0.05）。
表 1 各组小鼠给药处理表
Table 1. Dosing and treatment of mice in each group
序号 组别 是否注射环磷酰胺 第11~20 d灌胃 1 正常组 否 去离子水 2 模型对照组 是 去离子水 3 低剂量组 是 0.5 mg/kg 4 中剂量组 是 5 mg/kg 5 高剂量组 是 20 mg/kg 
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表 2 AGS对RAW264.7吞噬能力的影响
Table 2. Effects of AGS on the phagocytosis of RAW264.7 cells
组别 吸光度 正常组 0.282±0.014 200（μg/mL） 0.329±0.018* 500（μg/mL） 0.321±0.006* 1000（μg/mL） 0.320±0.007* LPS 0.338±0.015* 注：*表示与正常组相比，差异显著（P<0.05）。
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表 4 AGS对免疫低下小鼠碳廓清指数、吞噬指数的影响
Table 4. Effect of AGS on carbon clearance index and phagocytic index in immunocompromised mice
组别 碳廓清指数 吞噬指数 正常组 0.0689±0.0019 0.3551±0.0179 模型组 0.0199±0.0015* 0.0993±0.0119* 低剂量组 0.0210±0.0020* 0.1110±0.0116* 中剂量组 0.0339±0.0030*# 0.1888±0.0069*# 高剂量组 0.0409±0.0014*# 0.2489±0.0266*#
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表 5 AGS对小鼠脾脏T淋巴细胞亚群的影响
Table 5. Effect of AGS on T lymphocyte subsets of spleen in mice
组别 CD4+（%） CD8+（%） CD4+/CD8+ 正常组 72.33±0.40 21.63±0.64 3.35±0.11 模型组 56.17±0.57* 35.80±0.26* 1.57±0.02* 低剂量组 58.10±1.21* 27.80±0.87*# 2.09±0.11*# 中剂量组 66.23±0.40*# 22.13±0.57# 2.99±0.09*# 高剂量组 58.43±0.32*# 29.50±0.79*# 1.98±0.04*#
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