黑蒜提取物的抗氧化性与抗炎活性比较研究

目的对分离纯化黑蒜多糖的过程中得到6种黑蒜提取物,进行成分分析,并比较研究它们的抗氧化活性和抗炎活性。方法黑蒜经热水提取获得黑蒜水提物(hot water extract,HWE),经乙醇分离得到乙醇上清(ethanol supernatant extract,ESE)和沉淀(ethanol precipitate extract,EPE)提取物,并再经脱蛋白和乙醇分离得到脱蛋白的乙醇上清(deproteinized ethanol supernatant extract,d ESE)和沉淀(deproteinized ethanol precipitate extract,d EPE)提取物,再经DEAE 52纤维素层析纯化得到多糖提取物(purified polysaccharide extract,PPE)。首先采用薄层层析法分析了6种黑蒜提取物的单糖组成,还测定了总糖含量、还原糖含量、硫含量。再比较分析了6种多糖提取物的抗氧化与抗炎活性。结果 6种黑蒜提取物对DPPH自由基和ABTS自由基均具有良好的清除作用。6种黑蒜提取物对脂多糖(lipopoly saccharides,LPS)(20?g/mL)所致的RAW264.7细胞生长抑制均有显著性保护作用(P0.05)。6种黑蒜提取物对LPS(1?g/mL)诱导RAW264.7细胞NO,IL-1?,IL-6和TNF-?等细胞炎性因子的释放均有显著的抑制作用(P0.05)。结论黑蒜多糖提取物具有良好的抗氧化作用和抗炎活性,研究结果为黑蒜的进一步研发提供了理论依据和实际参考价值。

Antioxidant and anti-inflammatory activities of black garlic extracts

Objective Composition, antioxidant and anti-inflammatory activities of six black garlic extracts were assessed and compared. Methods Black garlics were prepared and processed by hot water extraction, ethanol separation, and anion-exchange purification. Six extracts were obtained, including HWE (Hot water extract), ESE (Ethanol supernatant extract), EPE (Ethanol precipitate extract), dESE (Deproteinized ethanol supernatant extract), dEPE (Deproteinized ethanol srecipitate extract), PPE (Purified polysaccharide extract). The monosaccharide type, total sugar content, reducing sugar content, sulfite content were analyzed by thin layer chromatography(TLC), phenol-sulfuric method, 3, 5-dinilrosalicylic acid method, brilliant green color-fading method. Antioxidant activities were studied by DPPH and ABTS free radical scavenging assay. Griess assay and ELISA were used to measure the levels of NO and inflammatory cytokines (IL-1beta, IL-6 and TNF-alpha), respectively. Results Six black garlic extracts were composed mainly of fructose, and their total sugar contents were in the order of PPE>EPE, dEPE>HWE>ESE, dESE, reducing sugar contents in the order of EPE, dEPE>HWE>ESE, dESE>PPE, sulfite contents in the order of HWE>ESE, dESE>EPE, dEPE>PPE. All six black garlic extracts had good DPPH and ABTS free radical scavenging activities, which were in the order of HWE>PPE>EPE, dEPE>ESE, dESE. All extracts protected RAW264.7 cells from LPS (20 ?g/mL) induced cell growth inhibition, which were in the order of HWE, PPE>EPE, dEPE>ESE, dESE. All extracts inhibited LPS(1 ?g/mL)-induced release of NO, IL-1beta, IL-6 and TNF-alpha of RAW264.7 cells, which were in the order of HWE>PPE>EPE, dEPE>ESE, dESE. Conclusion All six black garlic exhibited good antioxidant and anti-inflammatory activities. Our results provide scientific basis for the further exploitation of black garlic functional products.