CAJ | 학술논문

目的研究15 Hz、1 mT脉冲电磁场对骨髓间充质干细胞I型胶原表达的影响及其机制.方法体外培养SD大鼠骨髓间充质干细胞,取第3代细胞,用15 Hz、1 mT脉冲电磁场每日刺激8 h.用半定量逆转录聚合酶链反应(RT-PCR)检测I型胶原mRNA的表达,用酶联免疫吸附测定(ELISA)和细胞免疫组织化学等方法检测I型胶原的表达;加入环磷腺苷依赖的蛋白激酶A(cAMP-PKA)通路抑制剂和激动剂,并用ELISA和免疫组化的方法观察阻断和促进cAMP-PKA通路后脉冲电磁场对I型胶原表达的影响.结果 15 Hz、1 mT的脉冲电磁场每日刺激8 h可明显促进骨髓间充质干细胞I型胶原的表达(P<0.05),加入cAMP-PKA通路抑制剂H-89后,脉冲电磁场促进I型胶原表达效应明显减弱,加入cAMP-PKA通路激动剂8-Br-cAMP后脉冲电磁场促进的I型胶原表达效应明显增强.结论 15 Hz、1 mT脉冲电磁场可促进骨髓间充质干细胞I型胶原的表达,脉冲电磁场促进I型胶原表达的效应同cAMP-PKA通路密切相关.
목적 연구15 Hz、1 mT맥충전자장대골수간충질간세포I형효원표체적영향급기궤제.방법 체외배양SD대서골수간충질간세포,취제3대세포,용15 Hz、1 mT맥충전자장매일자격8 h.용반정량역전록취합매련반응(RT-PCR)검측I형효원mRNA적표체,용매련면역흡부측정(ELISA)화세포면역조직화학등방법검측I형효원적표체;가입배린선감의뢰적단백격매A(cAMP-PKA)통로억제제화격동제,병용ELISA화면역조화적방법관찰조단화촉진cAMP-PKA통로후맥충전자장대I형효원표체적영향.결과 15 Hz、1 mT적맥충전자장매일자격8 h가명현촉진골수간충질간세포I형효원적표체(P<0.05),가입cAMP-PKA통로억제제H-89후,맥충전자장촉진I형효원표체효응명현감약,가입cAMP-PKA통로격동제8-Br-cAMP후맥충전자장촉진적I형효원표체효응명현증강.결론 15 Hz、1 mT맥충전자장가촉진골수간충질간세포I형효원적표체,맥충전자장촉진I형효원표체적효응동cAMP-PKA통로밀절상관.
Objective To study the influence of pulsed electromagnetic fields on the expression of type I collagen by bone marrow mesenchymal stem cells and it's mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro.The third passage cells were harvested and exposed to pulsed electromagnetic fields(PEMFs)at 15 Hz and 1 mT 8 h/d for 3 days.A semi-quantitative RT-PCR technique was used to measure the type I collagen mRNA expression;ELISA and immunohistochemitistry techniques were used to measure type I collagen expression.Inhibitors and promoters of the cAMP-dependent protein kinase A(cAMP-PKA)pathway were added.After the cAMP-PKA pathway had been inhibited or promoted,the effects of the PEMF on type I collagen expression were measured again using ELISA and immunohistoehemistry.Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type I collagen(P≤0.01)in comparison with the controls. The type I collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added,and raised when the promoter 8-Br-cAMP was added.Conclusion PEMFs at 15 Hz 1 mT can promote type I collagen expression of bone marrow mesenchymal stem cells.and the effect is correlated with the cAMP-PKA pathway.