灵芝多糖对小鼠T细胞胞浆游离Ca~(2+)浓度和胞内pH的影响

目的通过考察灵芝多糖（ＧＬＰ）对免疫细胞信号转导过程的影响，探讨ＧＬＰ免疫调节作用机制。方法采用激光扫描共聚焦显微镜（ＬＳＣＭ）技术，动态监测ＧＬＰ均一体组分ＧＬＢ７对小鼠Ｔ细胞胞浆游离Ｃａ２＋浓度（［Ｃａ２＋）和胞内 ｐＨ（［ｐＨ］ｉ）的影响。结果 ＧＬＢ７（２０ｍｇ·Ｌ－１）引起小鼠 Ｔ细胞中［Ｃａ２＋］；和［ｐＨ］；明显升高，１ｍｉｎ即分别升高至３３４．７０％±１６，４％（ｎ＝３）、１７１．６％ ± １０．４％（ｎ＝３），之后一直分别维持在该平台期，至 １０ ｍｉｎ仍维持高峰水平。加入ＥＣＴＡ和 ｖｅｒａｐａｍｉｌ处理后， １０ ｍｉｎ ＧＬＢ７ 引起 Ｔ细胞［Ｃａ２＋］；和［ｐＨ］；分别升高为 ２０２．４％± １３．６％（ｎ＝３）。１４０．２％±７．８％（ｎ＝３），与正常对照组比较差异有显著性（Ｐ＜０．０1）。以 ＥＧＴＡ和 ｖｅｒａｐａｍｉｌ处理后，再分别以 ｈｅｐ－ｅｒｉｎ和 ｐrocaine处理细胞 １０ ｍｉｎ，之后以 ＧＬＢ７刺激Ｔ细胞，１０ｍｉｎ［Ｃａ２＋］ｉ值分别为１５１．５％±９．４％（ｎ＝３）、１４３．２％± ８．１ ％（ ｎ＝ ３），与 ＥＧＴＡ和 ｖｅｒａｐａｍｉｌ处理组相比差异有显著性（ Ｐ＜ ０．０１）。同时以

Effect of Ganoderma polysacchride on intracellular free calcium and intracellular pH in murine T cells

AIM　To extend the approach of the act ion of ganoderma polysacchride on intracellular signal events in T cells. METHODS　Laser scanning confocal microscope imaging of the calcium and p H fluorescent indicator dye Fluo-3/AM and SNARF-1/AM were used to determine th e kinetic changes of ［Ca2+］i and ［pH］i in murine T cells induced b y a ganoderma polysacchride,designated GLB7. RESULTS　It was fou nd that GLB7(20 mg*L-1) could increase ［Ca2+］i and ［pH］i at 1 min were 334.7%±16.4%(n=3)、171.6%±10.4%(n=3) respectively. T he increase in ［Ca2+］i induced by GLB7 was due to the influx of extr acellular Ca2+ and intracellular Ca2+ release through both IP3-se nsitive and IP3-insensitive Ca2+ stores, and increase in ［pH］i indu ced by GLB7 was relative to Na+/H+ exchange systems and ［Ca2+］i. GLB 7 did not influence ［Ca2+］i and ［pH］i in murine T cells induced by Con A(3 mg*L-1). CONCLUSION　Stimulation of the increase in ［Ca2+］i and ［pH］i may be an important channel for gano derma polysacchrides to achieve their pharmacological actions.