前列腺素E2调节大鼠成骨细胞OCIL基因表达的信号通路研究

探讨前列腺素E2(PGE2)对大鼠成骨细胞破骨细胞抑制性凝集素(osteoclast inhibitory lectin OCIL)mRNA表达的调节及其信号转导机制.培养大鼠原代成骨细胞和UMRl06成骨细胞样细胞,采用不同浓度的PGE2和不同的信号通路调节剂干预细胞后,提取细胞总RNA,采用实时荧光定量PCR检测OCILmRNA的表达水平.PGE2、蛋白激酶A(PKA)激动剂福司可林、db-cAMP和钙离子载体A23187均促进OCIL mRNA表达,OCIL mRNA最大上调幅度分别为对照组的2.38倍、4.2倍、4.5倍和5.1倍(均P＜0.01).蛋白激酶C(PKC)激动剂PMA下调OCILmRNA表达约50%(P＜0.01).PKA抑制剂KT-5720、钙通道阻断剂维拉帕米、钙调蛋白抑制剂W7和丝裂原活化蛋白激酶(MAPK)阻断剂PD98059分别下调PGE2诱导的OCIL mRNA表达约56%、40%、65%和60%(均P＜0.01).PKC抑制剂白屈菜红碱促进PGE2诱导的OCIL mRNA表达约30%(P＜0.05).PKA、MAPK和Ca2+/钙调蛋白信号通路介导了PCE2诱导的大鼠成骨细胞OCIL基因表达.

Abstract：

To investigate the regulation of osteoclast inhibitory lectin (OCIL) mRNA expression by prostaglandin E2 ( PGE2 ) in rat osteoblastic cells and the involved signaling pathways. Rat primary osteoblasts and UMR106 osteoblast-like cells were cultured and treated with various doses of PGE2 or regulators of different signaling pathways for different periods of time, the cells were then harvested at indicated dates. Total RNA were isolated and OCIL mRNA expression were studied by real-time PCR. PGE2, Forskolin, db-cAMP, and A23187 increased OCIL mRNA by 2. 38 fold,4. 2 fold,4. 5 fold, and 5. 1 fold ( all P＜0. 01 ) respectively, while PMA downregulated OCIL mRNA expression by 50% ( P＜0. 01 ). KT-5720, verapamil, W7, and PD98059 downregulated PGE2 induced OCIL mRNA expression by 56%, 40%, 65%, and 60%, respectively( all P＜0. 0l ). While chelerythrine enhanced PGE2 induced OCIL mRNA expression by 30% ( P＜0. 05 ). PGE2 up-regulated the expression of OCIL in rat osteoblastic cells via PKA, MAPK, and Ca2+/Calmodulin signaling pathways.

Regulation of osteoclast inhibitory lectin ( OCIL ) expression by prostaglandin E2 in rat osteoblastic cells

To investigate the regulation of osteoclast inhibitory lectin (OCIL) mRNA expression by prostaglandin E2 ( PGE2 ) in rat osteoblastic cells and the involved signaling pathways. Rat primary osteoblasts and UMR106 osteoblast-like cells were cultured and treated with various doses of PGE2 or regulators of different signaling pathways for different periods of time, the cells were then harvested at indicated dates. Total RNA were isolated and OCIL mRNA expression were studied by real-time PCR. PGE2, Forskolin, db-cAMP, and A23187 increased OCIL mRNA by 2. 38 fold,4. 2 fold,4. 5 fold, and 5. 1 fold ( all P＜0. 01 ) respectively, while PMA downregulated OCIL mRNA expression by 50% ( P＜0. 01 ). KT-5720, verapamil, W7, and PD98059 downregulated PGE2 induced OCIL mRNA expression by 56%, 40%, 65%, and 60%, respectively( all P＜0. 0l ). While chelerythrine enhanced PGE2 induced OCIL mRNA expression by 30% ( P＜0. 05 ). PGE2 up-regulated the expression of OCIL in rat osteoblastic cells via PKA, MAPK, and Ca2+/Calmodulin signaling pathways.