电磁场对骨髓间充质干细胞AgcⅠ、Ⅱ型胶原和Sox9等基因表达的影响

发布时间：2019-03-30 13:14

【摘要】：目的探讨15Hz、1mT电磁场对骨髓间充质干细胞成软骨分化指标聚集蛋白聚糖(AgcI),II型胶原和Sox9等的影响及初步机制。 方法体外培养SD大鼠骨髓间充质干细胞,取第3代细胞,用15Hz、1mT电磁场(EMFs)刺激,8h/d。采用半定量逆转录聚合酶链反应(RT-PCR)法检测甲状旁腺激素激素受体相关肽(PTHrp)、Agc I、II型胶原和Sox9等mRNA的表达,用Western Blot检测II型胶原蛋白的表达。加入解蛋白激酶A(PKA)抑制剂H-89和蛋白激酶C(PKC)抑制剂Go-6976,并再次用RT-PCR法和Western Blot观察EMFs对Agc I、II型胶原和Sox9等mRNA的表达的影响。 结果EMFs促进骨髓间充质干细胞PTHrp、Agc I、II型胶原和Sox9等mRNA的表达,促进II型胶原蛋白的表达;在加入H-89(10μm)和Go-6976(12μm)后EMFs的这种促进Agc I和II型胶原的效应就明显减弱,但Sox9的表达不受影响。 结论EMFs促进骨髓间充质干细胞成软骨分化指标AgcI、II型胶原和Sox9等mRNA的表达,这种促进作用同PKA和PKC通路有联系。
[Abstract]:Aim to investigate the effects of 15Hz and 1mT electromagnetic fields on the aggregation of proteoglycan (AgcI), type II collagen and Sox9 in chondrogenic differentiation of bone marrow mesenchymal stem cells (MSCs). Methods SD rat bone marrow mesenchymal stem cells were cultured in vitro. The third generation cells were stimulated with 15Hz, 1mT electromagnetic field (EMFs) for 8 h. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of type II collagen and Sox9, and Western Blot was used to detect the expression of type II collagen in parathyroid hormone receptor-related peptide (PTHrp), Agc I,. The effects of EMFs on the expression of Agc I, type II collagen, Sox9 and other mRNA were observed by RT- PCR and Western Blot. The expression of Agc I, collagen II and Sox9 were observed by RT- PCR and Western Blot. Results EMFs promoted the expression of mRNA such as PTHrp,Agc I, type II collagen and Sox9, and promoted the expression of type II collagen in bone marrow mesenchymal stem cells. The effect of EMFs on Agc type I and type II collagen was significantly decreased after addition of Go-6976 (12 渭 m) and 10 渭 m (10 渭 m), but the expression of Sox9 was not affected by the addition of Hx89 (10 渭 m) and Agc (12 渭 m). Conclusion EMFs promotes the expression of mRNA such as AgcI, type II collagen and Sox9, which is related to PKA and PKC pathway.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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