本文選題：降鈣素基因相關(guān)肽　切入點(diǎn)：成骨細(xì)胞　出處：《華西口腔醫(yī)學(xué)雜志》2017年02期 　論文類(lèi)型：期刊論文

【摘要】：目的研究血清饑餓條件下降鈣素基因相關(guān)肽(CGRP)對(duì)小鼠MC3T3-E1成骨細(xì)胞凋亡、自噬的影響以及二者之間的關(guān)系,以進(jìn)一步明確CGRP對(duì)成骨細(xì)胞的保護(hù)機(jī)制。方法體外培養(yǎng)小鼠MC3T3-E1成骨細(xì)胞。采用流式細(xì)胞術(shù)和蛋白質(zhì)印跡檢測(cè)正常血清、無(wú)血清(血清饑餓)、3-MA預(yù)處理+血清饑餓培養(yǎng)的成骨細(xì)胞的凋亡和微管相關(guān)蛋白1輕鏈3(LC3)蛋白的表達(dá)。采用蛋白質(zhì)印跡檢測(cè)正常血清、血清饑餓、血清饑餓+不同濃度(10~(-10)、10~(-9)、10~(-8)、10~(-7) mol·L~(-1))CGRP培養(yǎng)的成骨細(xì)胞的LC3和P62蛋白表達(dá);采用蛋白質(zhì)印跡檢測(cè)正常血清、血清饑餓、血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)不同時(shí)間(2、6、12、24、48、72 h)的成骨細(xì)胞的LC3蛋白表達(dá),流式細(xì)胞數(shù)檢測(cè)細(xì)胞凋亡,MDC染色檢測(cè)細(xì)胞自噬泡。采用流式細(xì)胞術(shù)檢測(cè)正常血清、血清饑餓、血清饑餓+10~(-8) mol·L~(-1) CGRP、3-MA預(yù)處理+血清饑餓、3-MA預(yù)處理+血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)24 h的成骨細(xì)胞的凋亡。結(jié)果血清饑餓培養(yǎng)時(shí)成骨細(xì)胞的LC3Ⅱ蛋白表達(dá)及細(xì)胞凋亡較正常血清時(shí)增加,3-MA預(yù)處理+血清饑餓培養(yǎng)時(shí)成骨細(xì)胞的凋亡較血清饑餓時(shí)增加(P0.01)。與正常血清相比,血清饑餓、血清饑餓+CGRP培養(yǎng)時(shí)LC3Ⅱ蛋白表達(dá)增加,P62蛋白表達(dá)降低,以血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)24 h時(shí)LC3Ⅱ/Ⅰ比值最高;血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)能抑制成骨細(xì)胞的凋亡,促進(jìn)自噬泡的合成。3-MA預(yù)處理后MC3T3-E1成骨細(xì)胞凋亡增加,CGRP部分逆轉(zhuǎn)3-MA預(yù)處理所增加的成骨細(xì)胞凋亡。結(jié)論 CGRP能夠增強(qiáng)血清饑餓下MC3T3-E1成骨細(xì)胞的自噬活性,并可能通過(guò)促進(jìn)自噬抑制成骨細(xì)胞的凋亡。
[Abstract]:Objective to study the effect of calcitonin gene-related peptide (CGRP) on apoptosis and autophagy of MC3T3-E1 osteoblasts in mice under serum starvation. Methods Mouse MC3T3-E1 osteoblasts were cultured in vitro. Normal serum was detected by flow cytometry and Western blotting. Apoptosis of osteoblasts and expression of microtubule-associated protein 1 light chain 3LC3 protein in cultured osteoblasts pretreated with serum starvation 3-MA. Western blotting was used to detect normal serum and serum starvation. The expression of LC3 and P62 protein in osteoblasts cultured with different concentrations of serum starvation was detected by Western blot, and the expression of LC3 protein was detected by Western blot in osteoblasts cultured with normal serum, serum starvation and serum hungry 10 mol 路L -1) CGRP. Flow cytometry was used to detect apoptosis and MDC staining to detect autophagy. Flow cytometry was used to detect normal serum and serum starvation. The expression of LC3 鈪，

本文編號(hào)：1592141