電磁場對骨髓間充質干細胞AgcⅠ、Ⅱ型膠原和Sox9等基因表達的影響
發(fā)布時間：2019-03-30 13:14
【摘要】：目的探討15Hz、1mT電磁場對骨髓間充質干細胞成軟骨分化指標聚集蛋白聚糖(AgcI),II型膠原和Sox9等的影響及初步機制。 方法體外培養(yǎng)SD大鼠骨髓間充質干細胞,取第3代細胞,用15Hz、1mT電磁場(EMFs)刺激,8h/d。采用半定量逆轉錄聚合酶鏈反應(RT-PCR)法檢測甲狀旁腺激素激素受體相關肽(PTHrp)、Agc I、II型膠原和Sox9等mRNA的表達,用Western Blot檢測II型膠原蛋白的表達。加入解蛋白激酶A(PKA)抑制劑H-89和蛋白激酶C(PKC)抑制劑Go-6976,并再次用RT-PCR法和Western Blot觀察EMFs對Agc I、II型膠原和Sox9等mRNA的表達的影響。 結果EMFs促進骨髓間充質干細胞PTHrp、Agc I、II型膠原和Sox9等mRNA的表達,促進II型膠原蛋白的表達;在加入H-89(10μm)和Go-6976(12μm)后EMFs的這種促進Agc I和II型膠原的效應就明顯減弱,但Sox9的表達不受影響。 結論EMFs促進骨髓間充質干細胞成軟骨分化指標AgcI、II型膠原和Sox9等mRNA的表達,這種促進作用同PKA和PKC通路有聯(lián)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.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R329
[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.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R329
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