前列腺素E2对成骨细胞微丝强度的调控作用  

Prostaglandin E2 regulated F-actin intensity in osteoblasts

作　　者：龚啸元[1] 王彬[1] 易彩霞[1] 潘君[1] 

机构地区：[1]重庆大学生物工程学院,400044

出　　处：《中华实验外科杂志》2014年第2期365-367,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（11272366、10972243）

摘　　要：目的 观察前列腺素E2（PGE2）在受到流体剪应力（FSS）加载后的MC3T3-E1成骨细胞株中对微丝强度的调控作用,并且探讨相关机制.方法 使用荧光标记法对经过FSS加载以及空白试剂、16,16-二甲基前列腺素E2（dmPGE2）、dmPGE2＋蛋白激酶A肽抑制剂（PKI）、8溴-环单磷酸腺苷（8Br-cAMP）、8Br-cAMP＋ PKI处理后的MC3T3-E1细胞中微丝进行染色处理;并通过图像处理软件（Volocity）对细胞中连续微丝的比例进行量化,检测dmPGE2对细胞中微丝强度的调控作用以及蛋白激酶A（PKA）信号通路的参与作用.结果 相对于空白对照组,dmPGE2显著降低了FSS加载后MC3T3-E1细胞中的微丝的强度.PKA信号通路的激活剂8Br-cAMP在一定程度上模拟了dmPGE2对微丝的作用;PKA信号通路阻滞剂PKI削弱了dmPGE2的作用.结论 dmPGE2能够促进FSS加载后MC3T3-E1细胞中微丝强度向静态水平的恢复;并且发现PKA信号通路参与此调控过程.Objective To explore the regulatory effect of 16,16-dimethyl prostaglandin E2 （dmPGE2） on fluid shear stress （FSS） loaded MC3T3-E1 osteoblastic cells and the underlying mechanism.Methods F-actin fluorescence staining and imaging quantification for continuous F-actin in 20 min FSS plus 15 min vehicle,dmPGE2,dmPGE2 ＋ protein kinase inhibitor （PKI）,8Br-cyclic adenosine monophosphate （8Br-cAMP）,8Br-cAMP ＋ PKI treated MC3T3-E1 cells were examined,which allowed us to analysis the regulatory effect of dmPGE2 on F-actin intensity,and the involvement of PKA pathway during dmPGE2 treatment.Results Compared with vehicle group,dmPGE2 significantly decreased the intensity of F-actin in FSS loaded MC3T3-E1 cells.8Br-cAMP,the PKA pathway activator mimicked the effect of dmPGE2 ; and PKI,the PKA pathway inhibitor attenuated the effect of dmPGE2.Conclusion Our study demonstrated that dmPGE2 was able to recover the F-actin intensity to the static level in FSS loaded MC3T3-E1 cells.PKA pathway was involved during this process.These findings provided a cellular mechanism by which PGE2 increases bone formation as shown in vivo,suggesting that PGE2 can be a potential target for osteoporosis related treatments.

关 键 词：成骨细胞 前列腺素E2 细胞骨架 流体剪应力 

分 类 号：R341[医药卫生—基础医学]