载脂蛋白D激活Wnt/β-catenin信号通路参与牙髓细胞增殖分化过程  

Involvement of apolipoprotein D in the proliferation and differentiation of dental pulp cells by activating Wnt/β-catenin signaling pathway

作　　者：林健生 孔令佳 鄂佳 王利娜 姚志文 LIN Jian-sheng;KONG Ling-jia;E Jia;WANG Li-na;YAO Zhi-wen(Department of Stomatology,Shenzhen Bao′an District Central Hospital,Guangdong Province,Shenzhen 518100,China)

机构地区：[1]广东省深圳市宝安区中心医院口腔科,广东深圳518100

出　　处：《河北医科大学学报》2024年第7期850-854,共5页Journal of Hebei Medical University

基　　金：广东省自然科学基金-面上项目(2020A1515011105);深圳市宝安区科技计划-基础研究项目(20210513212310001)。

摘　　要：目的 探究载脂蛋白D(apoliprotein D,APOD)激活果蝇无翅基因蛋白/β-连锁蛋白(Wnt/β-catenin)信号通路参与牙髓细胞(dental pulp cells, DPCs)增殖分化过程。方法 体外培养人牙髓细胞(human dental pulp cells, HDPCs),设置空白组、APOD低剂量组、APOD高剂量组和si-APOD组。空白组于DMEM培养基培养,APOD低剂量组向培养基中加入2 nmol/L APOD,APOD高剂量组向培养基中加入32nmol/LAPOD,si-APOD组采用APOD si-RNA转染HDPCs细胞。采用RT-qPCR检测APOD mRNA表达水平,CCK-8检测细胞增殖能力,碱性磷酸酶(alkaline phosphatase, ALP)活性检测成骨分化能力,Western Blot检测Wnt/β-catenin通路蛋白表达量。结果 与空白组比较,APOD低剂量组、APOD高剂量组APOD mRNA水平及细胞活力升高,7、14 d的ALP活性升高,Wnt5a、β-catenin蛋白上调(P<0.05),si-APOD组APOD mRNA水平,细胞活力降低,7、14 d的ALP活性降低,Wnt5a、β-catenin蛋白下调(P<0.05);与APOD低剂量组比较,APOD高剂量组APOD mRNA水平及细胞活力升高,7、14 d的ALP活性升高,Wnt5a、β-catenin蛋白上调(P<0.05),si-APOD组APOD mRNA水平及细胞活力降低,7、14 d的ALP活性降低,Wnt5a、β-catenin蛋白下调(P<0.05);与APOD高剂量组比较,si-APOD组APOD mRNA水平及细胞活力降低,7、14 d的ALP活性降低,Wnt5a、β-catenin蛋白下调(P<0.05)。结论 APOD能够促进HDPCs细胞的增殖分化,其作用机制可能与激活Wnt/β-catenin信号通路有关。Objective To investigate the involvement of apolipoprotein D(APOD)in the proliferation and differentiation of dental pulp cells(DPCs)by activating Wnt/β-catenin signaling pathway in Drosophila melanogaster.Methods Human dental pulp cells(HDPCs)were cultured in vitro and divided into blank group,low-dose APOD group,high-dose APOD group and si-APOD group.Cells in the blank group were cultured in DMEM medium without any treatment.APOD 2 nmol/L and 32 nmol/L were added to the low-dose APOD group and high-dose APOD group,respectively.HDPCs cells in the si-APOD group were transfected with APOD si-RNA.RT-qPCR was used to detect the expression of APOD mRNA,and CCK-8 was used to detect cell proliferation.Alkaline phosphatase(ALP)activity was used to detect osteogenic differentiation,and Western Blot was used to detect Wnt/β-catenin pathway protein expression.Results Compared with blank group,APOD mRNA level and cell viability were increased,ALP activity was increased at 7 d and 14 d,and Wnt5a andβ-catenin proteins were up-regulated in low-and high-dose APOD groups(P<0.05).In si-APOD group,APOD mRNA level and cell viability were decreased,ALP activity at 7 d and 14 d was decreased,and Wnt5a andβ-catenin proteins were down-regulated(P<0.05).Compared with the low-dose APOD group,the APOD mRNA level and cell viability were increased,the ALP activity was increased at 7 d and 14 d,and the Wnt5a andβ-catenin proteins were up-regulated in the high-dose group(P<0.05).In si-APOD group,APOD mRNA level and cell viability were decreased,ALP activity at 7 d and 14 d was decreased,and Wnt5a andβ-catenin protein were down-regulated(P<0.05).Compared with the high-dose APOD group,the mRNA level and cell viability of APOD were significantly decreased,the activity of ALP at 7 d and 14 d was decreased,and the proteins of Wnt5a andβ-catenin were down-regulated in si-APOD group(P<0.05).Conclusion APOD can promote the proliferation and differentiation of HDPCs cells,and its mechanism may be related to the activation of Wnt/β-catenin sign

关 键 词：牙髓 细胞增殖 载脂蛋白D类 

分 类 号：R322.41[医药卫生—人体解剖和组织胚胎学]