摘要
The present study aimed at isolating,purifying,characterizing the structure and estimating the anti-inflammatory activity of a polysaccharide named THSA which was obtained from Thamnolia subuliformis(Ehrh.) W.Culb(T.subuliformis).A newheteroglycan THSA was gained by hot water extraction and further continuously purification through Sephadex G-75 and Superdex 200 column chromatography.The structural characteristics of THSA were primarily investigated by molecular mass estimation,monosaccharide composition analysis and Fourier transforminfrared spectroscopy(FT-IR).The anti-inflammatory effects of THSA was also estimated in lipopolysaccharide(LPS)-stimulated mouse macrophages cell line RAW264.7 through the method of reverse transcription polymerase chain reaction(RT-PCR) and quantitative real-time PCR.The results indicate that THSA is composed of Glc,Gal,Man in the ratio 1:2.1:6.16,with average molecular weight of about 12.4 k Da,and significantly inhibits the mRNA expression of tumor necrosis factor alpha(TNF-α),IL-6 at100 or 1 000 μg/m L and increases the anti-inflammatory cytokine level of IL-10 at 100 μg/m L.It indicates that polysaecharide THSA has strong anti-inflammatory activity.
The present study aimed at isolating,purifying,characterizing the structure and estimating the anti-inflammatory activity of a polysaccharide named THSA which was obtained from Thamnolia subuliformis（Ehrh.） W.Culb（T.subuliformis）.A newheteroglycan THSA was gained by hot water extraction and further continuously purification through Sephadex G-75 and Superdex 200 column chromatography.The structural characteristics of THSA were primarily investigated by molecular mass estimation,monosaccharide composition analysis and Fourier transforminfrared spectroscopy（FT-IR）.The anti-inflammatory effects of THSA was also estimated in lipopolysaccharide（LPS）-stimulated mouse macrophages cell line RAW264.7 through the method of reverse transcription polymerase chain reaction（RT-PCR） and quantitative real-time PCR.The results indicate that THSA is composed of Glc,Gal,Man in the ratio 1：2.1：6.16,with average molecular weight of about 12.4 k Da,and significantly inhibits the mRNA expression of tumor necrosis factor alpha（TNF-α）,IL-6 at100 or 1 000 μg/m L and increases the anti-inflammatory cytokine level of IL-10 at 100 μg/m L.It indicates that polysaecharide THSA has strong anti-inflammatory activity.
基金
Fundamental Research Funds for the Central Universities,China(No.16D110523)
