高糖环境中程序性细胞死亡受体1抑制大鼠骨髓间充质干细胞的成骨分化  

Programmed death receptor 1 inhibits osteogenic differentiation of rat bone marrow mesenchymal stem cells in a high glucose environment

作　　者：韩念荣 黄异飞[1] 艾克热木·吾斯曼 刘岩路[1] 胡炜[1] Han Nianrong;Huang Yifei;Akram·Osman;Liu Yanlu;Hu Wei(Hospital of Traditional Chinese Medicine Affiliated to Xinjiang Medical University,Urumqi 830000,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学附属中医医院,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《中国组织工程研究》2025年第19期3961-3967,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金-杰出青年科学基金(2022D01E29),项目负责人:胡炜。

摘　　要：背景:程序性细胞死亡受体1(programmed death receptor-1,PD-1)在高糖环境下影响骨髓间充质干细胞成骨分化的作用机制尚不清楚。目的:探讨高糖环境中PD-1对大鼠骨髓间充质干细胞成骨分化的影响及其调控机制。方法:将大鼠骨髓间充质干细胞随机分为正常糖组(5.6 mmol/L)、高糖组(30 mmol/L)、PD-1过表达组、PD-1过表达空载组、PD-1敲低组、PD-1敲低空载组、PI3K/AKT通路抑制剂组(PD-1敲低+5μmol/L LY294002)。通过在高糖培养基中培养大鼠骨髓间充质干细胞来模拟体外糖尿病环境,采用qRT-PCR检测大鼠骨髓间充质干细胞中PD-1及其配体PD-L1和成骨标志物Runx2、OSX的mRNA表达,采用碱性磷酸酶染色和茜素红S染色观察成骨分化能力,采用CCK-8检测细胞增殖情况,采用Western blot检测PD-1、PD-L1、p-PI3K、p-AKT的蛋白表达。结果与结论:①高糖组PD-1及PD-L1表达显著高于正常糖组,高糖组骨髓间充质干细胞的成骨分化能力较正常糖组显著下降;②敲低PD-1表达可以促进骨髓间充质干细胞的成骨分化、增加细胞增殖活性,同时激活PI3K/AKT通路;③加入PI3K/AKT通路抑制剂LY294002后,骨髓间充质干细胞成骨分化能力显著下降。结果表明:PD-1依赖于PI3K/AKT信号通路抑制高糖环境下大鼠骨髓间充质干细胞的成骨分化。BACKGROUND:The mechanism of programmed death receptor-1(PD-1)effect on osteogenic differentiation of bone marrow mesenchymal stem cells in high glucose environment remains unclear.OBJECTIVE:To explore the effect of PD-1 on osteogenic differentiation of rat bone marrow mesenchymal stem cells in high glucose environment and its regulatory mechanism.METHODS:Rat bone marrow mesenchymal stem cells were randomly divided into normal glucose group(5.6 mmol/L),high glucose group(30 mmol/L),PD-1 overexpression group,PD-1 overexpression no-load group,PD-1 knockdown group,PD-1 knockdown no-load group,and PI3K/AKT pathway inhibitor group(PD-1 knockdown+5μmol/L LY294002).Rat bone marrow mesenchymal stem cells were cultured in high glucose to simulate the diabetic environment in vitro.The mRNA expression of PD-1 and ligand PD-L1 and the mRNA expression of osteogenic markers Runx2 and OSX in rat bone marrow mesenchymal stem cells were detected by qRT-PCR.The osteogenic differentiation ability was observed by alkaline phosphatase staining and alizarin red staining.Cell proliferation was detected by CCK-8 assay.The protein expressions of PD-1,PD-L1,p-PI3K,and p-AKT were detected by western blot assay.RESULTS AND CONCLUSION:(1)The levels of PD-1 and PD-L1 were significantly increased in the high glucose environment in vitro,and the osteogenic differentiation ability of bone marrow mesenchymal stem cells was inhibited in the high glucose environment.(2)Knockdown of PD-1 expression could promote osteogenic differentiation of bone marrow mesenchymal stem cells,increase cell proliferation activity,and activate the PI3K/AKT pathway.(3)After addition of PI3K/AKT pathway inhibitor LY294002,the ability of bone marrow mesenchymal stem cells to differentiate into osteoblasts decreased.The results show that PD-1 is dependent on the PI3K/AKT signaling pathway to inhibit osteogenic differentiation of rat bone marrow mesenchymal stem cells under high glucose environment.RESULTS AND CONCLUSION:(1)The levels of PD-1 and PD-L1 were significantly i

关 键 词：骨髓间充质干细胞 高糖 成骨分化 程序性细胞死亡受体1 PI3K AKT 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R587.1