CAJ | 학술논문

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.
목적 관찰전렬선소E2(PGE2)재수도류체전응력(FSS)가재후적MC3T3-E1성골세포주중대미사강도적조공작용,병차탐토상관궤제.방법 사용형광표기법대경과FSS가재이급공백시제、16,16-이갑기전렬선소E2(dmPGE2)、dmPGE2+단백격매A태억제제(PKI)、8추-배단린산선감(8Br-cAMP)、8Br-cAMP+ PKI처리후적MC3T3-E1세포중미사진행염색처리;병통과도상처리연건(Volocity)대세포중련속미사적비례진행양화,검측dmPGE2대세포중미사강도적조공작용이급단백격매A(PKA)신호통로적삼여작용.결과 상대우공백대조조,dmPGE2현저강저료FSS가재후MC3T3-E1세포중적미사적강도.PKA신호통로적격활제8Br-cAMP재일정정도상모의료dmPGE2대미사적작용;PKA신호통로조체제PKI삭약료dmPGE2적작용.결론 dmPGE2능구촉진FSS가재후MC3T3-E1세포중미사강도향정태수평적회복;병차발현PKA신호통로삼여차조공과정.
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.