Influences of Sulfated Polysaccharide from Pine (Pinus massoniana) Pollen on the Immunomodulatory Effects of B Lymphocytes in Mice

Abstract: We investigated the influences of a polysaccharide component D (PPM60-D) and its sulphate (SPPM60-D) from masson pine pollen on the proliferation， intracellular free calcium concentration ([Ca2+]i) and antibody production of B lymphocytes in mice. Polysaccharides (PPM60) were extracted from masson pine pollen with hot water and precipitated by 60% ethanol. PPM60-D was purified from PPM60 with Sephacryl S-400HR. Sulfated polysaccharides (SPPM60-D) was derivated by chlorosulfonic acid-pyridine method. B lymphocytes were separated through nylon wool column. [Ca2+]i of B lymphocytes were measured by fluorescence sepectrophotometer. The antibody production were measured by PFC and QHS methods. The results showed that SPPM60-D could more significantly improve the proliferation， [Ca2+]i and antibody production of B lymphocytes than PPM60-D (P<0.01). TAK-242， LY294002， U73122， low molecular heparin， verapamil and 2-APB could inhibit the increasement of [Ca2+]i in B lymphocytes which was induced by PPM60-D and SPPM60-D (P<0.05 or P<0.01). These suggested that SPPM60-D had higher activity than PPM60-D and one of the possible receptors of SPPM60-D was toll like receptor 4 (TLR4). We speculated that SPPM60-D could increase [Ca2+]i via Ca2+ release-activated Ca2+ (CRAC) channel which was activated by TLR4-PI3K-PLC-IP3R signaling pathway， so that it could improve the ability of proliferation and antibody production of B lymphocytes.

CSTR: 32200.14.cjcb.2014.04.0009