本文選題：降鈣素基因相關肽　切入點：成骨細胞　出處：《華西口腔醫(yī)學雜志》2017年02期 　論文類型：期刊論文

【摘要】：目的研究血清饑餓條件下降鈣素基因相關肽(CGRP)對小鼠MC3T3-E1成骨細胞凋亡、自噬的影響以及二者之間的關系,以進一步明確CGRP對成骨細胞的保護機制。方法體外培養(yǎng)小鼠MC3T3-E1成骨細胞。采用流式細胞術和蛋白質印跡檢測正常血清、無血清(血清饑餓)、3-MA預處理+血清饑餓培養(yǎng)的成骨細胞的凋亡和微管相關蛋白1輕鏈3(LC3)蛋白的表達。采用蛋白質印跡檢測正常血清、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)的成骨細胞的LC3和P62蛋白表達;采用蛋白質印跡檢測正常血清、血清饑餓、血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)不同時間(2、6、12、24、48、72 h)的成骨細胞的LC3蛋白表達,流式細胞數(shù)檢測細胞凋亡,MDC染色檢測細胞自噬泡。采用流式細胞術檢測正常血清、血清饑餓、血清饑餓+10~(-8) mol·L~(-1) CGRP、3-MA預處理+血清饑餓、3-MA預處理+血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)24 h的成骨細胞的凋亡。結果血清饑餓培養(yǎng)時成骨細胞的LC3Ⅱ蛋白表達及細胞凋亡較正常血清時增加,3-MA預處理+血清饑餓培養(yǎng)時成骨細胞的凋亡較血清饑餓時增加(P0.01)。與正常血清相比,血清饑餓、血清饑餓+CGRP培養(yǎng)時LC3Ⅱ蛋白表達增加,P62蛋白表達降低,以血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)24 h時LC3Ⅱ/Ⅰ比值最高;血清饑餓+10~(-8) mol·L~(-1) CGRP培養(yǎng)能抑制成骨細胞的凋亡,促進自噬泡的合成。3-MA預處理后MC3T3-E1成骨細胞凋亡增加,CGRP部分逆轉3-MA預處理所增加的成骨細胞凋亡。結論 CGRP能夠增強血清饑餓下MC3T3-E1成骨細胞的自噬活性,并可能通過促進自噬抑制成骨細胞的凋亡。
[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：1592141