基质细胞衍生因子1α通过Akt信号通路抑制软骨细胞凋亡并促进自噬  

Stromal Cell-Derived Factor 1αInhibits Chondrocyte Apoptosis and Promotes Autophagy Through the Akt Signaling Pathway

作　　者：李嘉舟 谢静 周学东[1] LI Jiazhou;XIE Jing;ZHOU Xuedong(State Key Laboratory of Oral Diseases&National Center for Stomatology&National Clinical Research Center for Oral Diseases,West China Hospital of Stomatology,Sichuan University,Chengdu 610041,China)

机构地区：[1]口腔疾病防治全国重点实验室国家口腔医学中心,国家口腔疾病临床医学研究中心,四川大学华西口腔医院,成都610041

出　　处：《四川大学学报（医学版）》2024年第1期53-59,共7页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81870754)资助。

摘　　要：目的探究基质细胞衍生因子1α(stromal cell-derived factor 1α,SDF-1α)对软骨细胞凋亡及自噬的影响及其潜在机制。方法从新生小鼠的膝关节中分离提取软骨细胞,以0(对照组)、50、100和200 ng/mL的SDF-1α刺激软骨细胞,CCK-8实验检测SDF-1α刺激24 h、48 h、72 h对软骨细胞活性的影响,划痕实验检测SDF-1α刺激12 h、24 h对软骨细胞迁移能力的影响,Western blot实验测定SDF-1α作用后软骨细胞内Akt信号通路相关蛋白表达变化。以0(对照组)、200 ng/mL的SDF-1α刺激软骨细胞,流式细胞术检测SDF-1α对软骨细胞凋亡的影响,透射电镜下检测SDF-1α对软骨细胞自噬的影响,并利用免疫荧光染色实验直观显示SDF-1α作用后软骨细胞内p-Akt蛋白表达及分布的差异。结果与对照组相比,50、100和200 ng/mL SDF-1α在各时点并不降低软骨细胞活性(P<0.01),且均能在24 h促进软骨细胞迁移(P<0.05)。Western blot实验结果显示,与对照组相比,50、100和200 ng/mL SDF-1α能够显著上调软骨细胞内p-Akt的蛋白表达量,Akt表达量未见明显差异。与对照组相比,流式细胞术示SDF-1α能够抑制软骨细胞凋亡(P<0.05),透射电镜观察到SDF-1α促进软骨细胞自噬(P<0.05),免疫荧光染色实验结果显示,p-Akt在软骨细胞的表达主要集中在细胞核周,SDF-1α作用后软骨细胞内p-Akt表达进一步在细胞核周部位增强。结论SDF-1α通过激活Akt信号通路,抑制软骨细胞凋亡,促进软骨细胞迁移和自噬。Objective To investigate the effects of stromal cell-derived factor 1α(SDF-1α)on the apoptosis and autophagy of chondrocytes and the underlying mechanisms.Methods Chondrocytes were isolated from the knee joints of neonatal mice.The chondrocytes were then stimulated with 0(the control group),50,100,and 200 ng/mL of SDF-1α.CCK-8 assay was performed to determine the effects of SDF-1αstimulation for 24 h,48 h,and 72 h on the viability of the chondrocytes.Wound healing assay was conducted to determine the effects of SDF-1αstimulation for 12 h and 24 h on chondrocyte migration.The changes in the expression of Akt signaling pathway proteins in chondrocytes were determined by Western blot assay.Chondrocytes were stimulated with 0(the control group)and 200 ng/mL of SDF-1α.Flow cytometry was performed to determine the effect of SDF-1αon the apoptosis of chondrocytes.Transmission electron microscope was used to examine the effect of SDF-1αon chondrocyte autophagy.Immunofluorescence staining assays were performed to visualize the differences in p-Akt expression and distribution in chondrocytes treated with SDF-1α.Results Compared with the control group,findings for the experimental groups showed that SDF-1αat the concentrations of 50,100,and 200 ng/mL did not decrease chondrocyte activity at any time point(P<0.01)and it consistently promoted chondrocyte migration at 24 h(P<0.05).Western blot results revealed that,in comparison to the control group,SDF-1αat concentrations of 50,100,and 200 ng/mL significantly up-regulated the protein expression of p-Akt in chondrocytes,while no significant difference in Akt expression was observed.Flow cytometry demonstrated that SDF-1αcould inhibit chondrocyte apoptosis(P<0.05)and transmission electron microscopic observation showed that SDF-1αpromoted chondrocyte autophagy(P<0.05).Immunofluorescence staining showed that the expression of p-Akt in chondrocytes was concentrated in the perinuclear area of the cells and this expression was further enhanced in the perinuclear area o

关 键 词：基质细胞衍生因子1Α 软骨细胞 凋亡 AKT 自噬 

分 类 号：R684.3[医药卫生—骨科学]