微米化保健食品配方免疫調節與抗腫瘤活性之研究

Abstract

本研究利用微米研磨技術針對人蔘 (Panax ginseng)、綠茶 (Camellia sinensis)、洋甘菊 (Camellia sinensis) 、金針菇 (Flammulina veluptipes) 等乾燥粉末調配得之組合性保健食品配方 J，處理得到微米化組合性保健食品配方 NJ，以 BALB/c 小鼠進行分析，餵食經過微米處理後之樣品與未經過微米處理之配方 J 其調節免疫與抑制小鼠肝腫瘤之活性之比較。 體內非特異性免疫調節活性結果顯示，餵食組合性保健食品配方J 高、中劑量 (分別為27.8 mg/mouse, 9.3 mg/mouse) 與微米化配方J (NJ) 高劑量可顯著提高小鼠脾細胞增生率 (p<0.05)；餵食配方 J 高劑量與微米配方 NJ 高劑量可顯著提升脾細胞 IFN-γ分泌量，而餵食配方 J 高中劑量可提高 IL-2分泌量 (p<0.05)；餵食配方 J 高中劑量可提高 NK 細胞毒殺活性 (p<0.05)；餵食配方 J 高中劑量與微米配方 NJ 高中劑量可顯著提升血液中單核球細胞吞噬能力 (p<0.05)；餵食配方 J 高劑量與微米配方 NJ 高中劑量可顯著提升小鼠血清中 IFN-γ 分泌量(p<0.05)；餵食配方 J 高中劑量與微米配方 NJ 高中劑量可顯著提升小鼠血清中總 IgG分泌量 (p<0.05)。由以上結果餵食組合性保健食品配方及其微米處理配方均具有調節非特異免疫反應之功效，但實驗數據顯示未經微米處理之配方 J 效果較佳。 體內 OVA 特異性免疫調節活性結果顯示，餵食配方 J 高劑量與微米配方 NJ 中劑量可顯著提高小鼠 OVA 特異性脾細胞增生率 (p<0.05)；餵食微米配方 NJ 高中劑量可顯著提昇脾細胞 OVA 特異性IFN-γ 分泌量 (p<0.05)，其中餵食 J 高劑量與微米配方 NJ 高中劑量亦可提高脾細胞 OVA 特異性 IL-2 分泌量 (p<0.05)。因此，餵食組合性保健食品配方及其微米處理配方具有調節 OVA 特異性免疫反應之功效。然而比較經過微米處理與否，配方 J 與經過微米處理配方 NJ在調節OVA特異性免疫之能力兩者並無顯著差異。 在抑制小鼠肝腫瘤試驗方面，以本實驗室純化所得之金針菇免疫調節蛋白 FIP-fve (0.2 mg/mouse) 作為正控制組，罹癌小鼠存活試驗顯示，腹腔注射ATCC BNL 1MEA.7R.1 肝癌細胞 (3×104 cell/mouse) 之BALB/c小鼠經餵食配方 J 高劑量與微米配方 NJ 高劑量，相較於未時對照組 (PBS)，可顯著延長小鼠存活時間 (p<0.05)。在非特異性抗腫瘤活性方面，結果顯示餵食配方 J 高中劑量與微米配方 NJ 中劑量可顯著活化腹腔巨噬細胞提升細胞激素 TNF-α 分泌量及 NO 之產生量；且小鼠腹腔巨噬細胞毒殺小鼠肝腫瘤細胞能力方面，餵食配方 J 高中劑量與微米配方 NJ 高中劑量與對照組相比皆具有顯著提升 (p<0.05)。在腫瘤特異性抗腫瘤活性方面結果顯示，餵食配方 J 高中劑量與微米配方 NJ 高中劑量均可顯著提高腫瘤特異性之小鼠脾細胞增生率 (p<0.05)；並顯著提升小鼠脾細胞之腫瘤特異性 IgG 分泌量 (p<0.05)；且小鼠脾臟細胞毒殺小鼠肝癌細胞能力，與對照組相比均具有顯著差異 (p<0.05)；餵食配方 J 高劑量與微米配方 NJ 高劑量可顯著提升小鼠脾臟細胞之腫瘤特異性 TNF-α 分泌量 (p<0.05)；餵食配方 J 高中劑量可顯著提升小鼠脾臟細胞之腫瘤特異性 IL-2 分泌量 (p<0.05)。然而比較經過微米處理與否，配方 J 與經過微米處理配方 NJ在調節腫瘤非特異性及腫瘤特異性抗腫瘤活性之能力兩者並無顯著差異，且均具有活化非特異性及腫瘤特異性之抑制小鼠肝腫瘤作用。

The objective of this study was to evaluate the immunomodulatory effects and anti-tumor effects of multi-ingredients health food formula J and micronized formula J (NJ). Results of the evaluation on non-specific immunomodulatory properties demonstrated that administration of formula J at high and medium doses (27.8 mg/mouse, 9.3 mg/mouse respectively) , as well as micronized formula J (NJ) at high doses (27.8 mg/mouse) significantly (p<0.05) enhanced the cell proliferation of mouse splenocytes. Administration of formula J and micronized formula NJ at high doses significantly (p<0.05) increased IFN-γ secretion by mouse splenocytes. Administration of formula J at high and medium doses significantly (p<0.05) increased the IL-2 secretion by mouse splenocytes. Administration of formula J at high and medium doses significantly (p<0.05) enhanced the NK cells activity of mouse splenocytes. Administration of formula J and micronized formula NJ (both at high and medium doses) significantly (p<0.05) enhanced the phagocytosis activity of monocytes. Administration of formula J (high dose) and micronized formula NJ (high and medium doses) significantly (p<0.05) increased the IFN-γ secretion in mouse sera. Administration of formula J and micronized formula NJ (both at high and medium doses) significantly (p<0.05) increased the total IgG secretion in mouse sera. These results showed that formula J was better then the micronized formula NJ on the non-specific immunomodulatory properties. Results of the evaluation on OVA-specific immunomodulatory properties showed that administration of formula J (high dose) and micronized formula NJ (medium dose) significantly (p<0.05) enhanced the OVA-specific cell proliferation of mouse splenocytes. Administration of micronized formula NJ at high and medium doses significantly (p<0.05) increased OVA-specific IFN-γ secretion by mouse splenocytes. Administration of formula J (high dose) and micronized formula NJ (high and medium doses) significantly (p<0.05) increased the OVA-specific IL-2 secretion by mouse splenocytes. The objective was to investigate the anti tumor effects and its related mechanisms of formula J. Administration of formula J (high dose) and micronized formula NJ (high dose) significantly (p < 0.05) increased the life span of ATCC BNL 1MEA.7R.1 hepatoma-bearing mice (3×104 cells/mouse) by 30.2％ and 21.5％, respectively. This result suggested that formula J and micronized formula NJ displayed activities to suppress hepatoma growth in vivo. The results of non-specific anti-tumor activities showed that administration of formula J (high and medium doses) and micronized formula NJ (medium dose) significantly (p < 0.05) stimulated mouse peritoneal macrophages to secret TNF-α and produce nitric oxide. Additionally, the tumoricidal activity of peritoneal cells was also significantly enhanced (p < 0.05). Administration of formula J and micronized formula NJ (both at high and medium doses) significantly (p < 0.05) enhanced the tumor-specific cell proliferation of hepatoma-bearing mice splenocytes, and also significantly (p < 0.05) enhanced the tumor-specific IgG secretion in serum. Administration of formula J and micronized formula NJ (both at high doses) significantly (p < 0.05) increased the tumor-specific TNF-α secretion by hepatoma-bearing mice splenocytes. Administration of formula J at medium dose significantly (p < 0.05) increased the tumor-specific IL-2 secretion by hepatoma-bearing mice splenocytes. The tumoricidal activities of splenocytes cells obtained from hepatoma-bearing mice fed the formulas J and NJ samples were also significantly (p < 0.05) enhanced. These results showed that formula J and micronized formula NJ samples carried out anti-tumor activity by activating both the non-specific and tumor-specific immunities of their hosts.