“南薯88”茎叶多糖的分离纯化,结构鉴定及其生物活性的研究

Isolation,Purification,Structure Identification And Biological Activity of Polysaccharides From Stems And Leaves of "Nanshu 88"

晒干的"南薯88"的茎叶,通过热水浸提,乙醇沉淀,DEAE-cellulose column柱层析分离纯化,得到"南薯88"多糖纯品(N88P-1)。通过红外光谱技术(IR),高效凝胶渗透色谱(HPGPC),高效液相色谱色谱(HPLC)和核磁共振谱(1 H NMR)等对"南薯88"多糖(N88P-1)的进行了结构鉴定;同时,对"南薯88"多糖(N88P-1)的抗氧化及免疫调控作用进行了初探。结果显示,"南薯88"多糖(N88P-1)的分子量为1 4328 D,红外光谱显示,在3 444.81 cm^(-1)出现O-H的特征伸缩振动峰,1 640.20 m^(-1)出现C=O的特征伸缩振动峰,~1H NMR谱显示δ4.93～δ4.86出现异头氢信号,高效液相色谱色谱显示N88P-1的单糖组成为1∶1的半乳糖和甘露糖。活性研究结果显示,一定范围内的N88P-1浓度可以清除ABTS^+自由基,IC_(50)值为0.48 mg/mL。N88P-1在0.25～3 mg/mL的浓度范围内与DPPH^-自由基的清除能力呈正相关,IC_(50)值为0.96 mg/mL。当N88P-1的浓度为3 mg/mL时,对ABTS^+和DPPH^-自由基的清除能力均达到最高值;免疫细胞增殖活性结果显示,N88P-1在20～80 mg/mL的浓度下,对B细胞和T细胞增殖效果为极显著(P<0.01);在10～40 mg/mL的浓度下,对巨噬细胞增殖效果为极显著(P<0.01),其中,N88P-1在40 mg/mL的浓度下对3种免疫细胞的增殖效果均达到最佳。

The"Nanshu 88"polysaccharide(N88P-1)is obtained from the dried stems and leaves of"Nanshu 88"by hot water extraction,ethanol precipitation,and then purified by DEAE-cellulose 52column chromatography.The structure of"Nanshu 88"pure polysaccharide(N88P-1)is identification by Infrared spectroscopy(IR),High Performance Gel Penetration Chromatography(HPGPC),High Performance Liquid Phase Chromatographic(HPLC)and nuclear magnetic resonance spectra(1H NMR and13C NMR).At the same time,the antioxidant and immune regulation effects of N88P-1 are also carries out.The results show that N88P-1 has molecular weight of 14 328D.The infrared spectra shows the characteristic peaks of O-H stretching vibration appears in 3 444.81cm^-1,and the characteristic peaks of C=O stretching vibration appears in 1 640.20 cm^-1.1H NMR spectra showed anomeric hydrogen signals atδ4.93~δ4.86.High performance liquid chromatography(HPLC)chromatography shows N88P-1 is composed of galactose and mannose with the ratio of 1∶1The activity study shows the N88P-1 has ability to clear the ABTS+free radical in a certain range of concentration,and the IC50value is 0.48 mg/mL.The DPPH-free radical scavenging activity of N88P-1 is positively correlated with concentration in the range of 0.25~3 mg/mL,the IC50value is0.96 mg/mL.When the concentration of N88P-1 is 3 mg/mL,the scavenging activity of ABTS+and DPPH-free radical both reach the highest value.The proliferation activity of the immune cells results show that the proliferation effect on B cells and T cells are very significant(P<0.01)in the concentration of 20~80 mg/mL;while the effect of macrophage proliferation was very significant(P<0.01)in the concentration of 10~40 mg/mL;It is worth noting that the proliferation effect of the three kinds of immune cells all reached the highest value in the concentration of 40 mg/mL.