中国药理学通报2012，Vol.28Issue(4)：478-482,5.DOI:10.3969/j.issn.1001-1978.2012.04.009
环磷腺苷通路对内毒素刺激的小胶质细胞分泌细胞因子的影响
The effect of cAMP signaling pathway on the expression of cytokines by the microglia stimulated with endotoxin
摘要
Abstract
Aim To observe the effect of protein kinase A (PKA) and Epac ( exchange proteins directly activated by cAMP)-the downstream effectors of cAMP onthe expression of inflammatory cytokines by the primary cultured microglia stimulated with endotoxin. Methods The primary cultured microglia cells from brains of new-born rats were embedded in the 24 pore plate then treated with different drugs as the following: group 1 in which microglias were incubated with 10% DMSO for 30 min, then HBSS for 30 min, finally HBSS for 24 h; group 2 with 10% DMSO and HBSS for 30 min respectively, group 3 with 10% DMSO and 100 nmol 6-Bnz-cAMP for 30 min respectively, group 4 with 10% DMSO and 50 nmol 8-pCPT-2'-O-Me-cAMP for 30 min respectively, group 5 with 50 nmol H-89 and 100 nmol 6-Bnz-cAMP for 30 min respectively, group 6 with 50 nmol H-89 and 50 nmol 8-pCPT-2'-O-Me-cAMP for 30 min respectively, finally 0. 5 (xg endotoxin for 24 h for group 2 to 6. The morphologic changes were observed with an inverted microscope 0, 3 h, 6 h and 24 h after endotoxin stimluation and the ratio of long axis of mi-croglia over short axis at 6 h was calcuated. The super-nate was collected at 24 h after endotoxin chanllenge and the levels of TNF-a, IL-1 p and IL-10 were meas-ured by ELISA. Results The morphology of microglia changed to bipolar rod shapes 3 h after endotoxin challenge , reached peak at 6 h and then returned to oval or amoeboid at 24 h. Only treatment with 6-Bnz-cAMP could significantly inhibit the morphologic change at 6 h, but pretreatment with H-89 completely reversed the effect of 6-Bnz-cAMP. Treatment with 6-Bnz-cAMP could significantly inhibited the overexpression of TNF-a and IL-1 (3 and enhance the secretion of IL-10, while these effects were completely reversed by H-89. Treatment with 8-pCPT-2'-O-Me-cAMP only slightly reduced the expression of TNF-a, meanwhile this effect was not influenced by H-89. Conclusion Compared with Epac, PKA shows more important effects on the expression of inflammatory cytokines by the primary cultured microglia stimulated with endotoxin.关键词
环磷腺苷/小胶质细胞/炎症细胞因子/蛋白激酶A/环腺苷酸直接活化的交换蛋白/内毒素Key words
cAMP/ microglia/ inflammatory cytokines/ PKA/ Epac/ endotoxin分类
医药卫生引用本文复制引用
刘健,谢军明,嵇晴,张利东,朱四海,李伟彦..环磷腺苷通路对内毒素刺激的小胶质细胞分泌细胞因子的影响[J].中国药理学通报,2012,28(4):478-482,5.基金项目
国家自然科学基金资助项目(No 30901399) （No 30901399）
中国博士后基金面上资助(No 20080440625) （No 20080440625）
