马尾松花粉硫酸酯化多糖调控脾细胞[Ca^(2+)]_i的研究

Influences of sulfated polysaccharide from pine (Pinus massoniana) pollen on the regulation of [Ca^(2+)]_i in splenocytes

目的探讨马尾松花粉多糖硫酸酯化物(SPPM60-A)对小鼠脾细胞内游离钙离子浓度([Ca2+]i)调控的机制。方法热水浸提醇沉淀法提取马尾松花粉粗多糖,SephacrylS-400HR分离纯化,氯磺酸-吡啶法进行硫酸酯化,荧光分光光度计测定脾淋巴细胞[Ca2+]i。结果 LPS与SPPM60-A都能促进脾淋巴细胞[Ca2+]i升高,与对照相比升高率分别为11.7%和15.4%(P<0.01)。TAK-242、LY294002、U73122、低分子肝素与维拉帕米均可以抑制[Ca2+]i的升高。结论推测硫酸酯化马尾松花粉多糖可通过TLR4-PI3K-PLC-IP3R信号通路促进脾淋巴细胞内[Ca2+]i升高。

Aim To study the influences of sulfated polysaccharides on the regulation of free calcium concentration（[Ca2 ＋]i） of splenocytes of mice in vitro and explore the mechanisms involved.Methods Polysaccharides were extracted from Pinus massoniana pollen with hot water.Polysaccharides were Separated and purified with Sephacryl S-400HR.Sulfated polysaccharides（SPPM60） were derivated by chlorosulfonic acid-pyridine method and the [Ca2 ＋]i of splenocytes were measured by fluorescence spectrophotometer.Results LPS and SPPM60-A increased[Ca2 ＋]icompared with the control in splenocytes at the rates 11.7%（P 0.01） and 15.4%（P 0.01） respectively.TAK-242,LY294002,U73122,low molecular weight heparin and verapamil significantly inhibited the increase of [Ca2 ＋]i induced by LPS and SPPM60-A.Conclusion Sulfated polysaccharides from pine pollen can increase [Ca2 ＋]i via TLR4-PI3K-PLC-IP3R signaling pathway in splenocytes.