miR-365通过靶向调控SIRT1对髓核细胞增殖、凋亡的影响  

The influence of miR-365 on proliferation and apoptosis of nucleus pulposus cells by targeted-regulating silencing information regulator factor 1

作　　者：艾则孜·艾海提[1] 许建[1] 郭瑞[1] Aizezi Aihaiti;XU Jian;GUO Rui(Dept of Orthopaedics,the People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院骨科,新疆乌鲁木齐830001

出　　处：《临床骨科杂志》2024年第4期590-595,共6页Journal of Clinical Orthopaedics

基　　金：新疆医科大学自治区研究生科研创新项目(编号:XJ2022G159)。

摘　　要：目的探讨miR-365对髓核细胞增殖、凋亡的影响及其与细胞沉默信息调节因子1(SIRT1)的靶向关系。方法选取人髓核细胞,正常培养的纳入对照组,用脂多糖(LPS)刺激细胞模拟椎间盘变性的纳入LPS组。将质粒转染细胞,然后用LPS处理,记为miR-365组、si-miR-365组、miR-NC组、NC组、SIRT1组、Vector组、si-SIRT1组、Scramble组、si-miR-365+si-SIRT1组、si-miR-365+Scramble组、miR-365+WT-SIRT1组、miR-NC+WT-SIRT1组、miR-365+MUT-SIRT1组及miR-NC+MUT-SIRT1组。检测LPS、miR-365、SIRT1对髓核细胞增殖、凋亡的影响,TargetScan在线网站预测miR-365与SIRT1结合位点,双荧光素酶报告实验验证miR-365与SIRT1的靶向关系。结果细胞上清液白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α含量及细胞miR-365水平LPS组均高于对照组(P<0.01),SIRT1蛋白、基因mRNA水平低于对照组(P<0.01)。LPS处理的髓核细胞增殖能力降低、凋亡增加。上调miR-365能够抑制LPS处理的髓核细胞的增殖活性、促进凋亡、降低SIRT1表达,下调miR-365可得到相反的结果。上调SIRT1能够促进LPS处理的髓核细胞的增殖活性、抑制凋亡,下调SIRT1可得到相反的结果。敲低SIRT1可逆转miR-365对髓核细胞增殖、凋亡的影响。miR-365负性调控SIRT1表达。结论miR-365通过靶向调控SIRT1而影响髓核细胞增殖、凋亡,为椎间盘退变的防治提供了新靶点。Objective To investigate the influence of miR-365 on proliferation and apoptosis of nucleus pulposus cells and its targeting relationship with cell silencing information regulator factor 1(SIRT1).Methods Human nucleus pulposus cells(HNPCs)were selected,and normal cultured cells were included in the control group.Cells stimulated with lipopolysaccharide(LPS)to simulate disc degeneration were included in the LPS group.The plasmid was transfected into cells and then treated with LPS,it was denoted as miR-365 group,si-miR-365 group,miR-NC group,NC group,SIRT1 group,Vector group,si-SIRT1 group,Scramble group,si-miR-365+si-SIRT1 group,si-miR-365+Scramble group,miR-365+WT-SIRT1 group,miR-NC+WT-SIRT1 group,miR-365+MUT-SIRT1 group and miR-NC+MUT-SIRT1 group.The effects of LPS,miR-365 and SIRT1 on proliferation and apoptosis of nucleus pulposus cells were detected.The TargetScan online website predicted the binding sites of miR-365 and SIRT1,and dual luciferase reporting experiments verified the targeting relationship between miR-365 and SIRT1.Results The contents of interleukin(IL)-6,IL-10,tumor necrosis factor(TNF)-αand miR-365 in LPS group were higher than those in control group(P<0.01),and the levels of SIRT1 protein and gene mRNA were lower than those in control group(P<0.01).LPS treated HNPC cell proliferation was decreased,and apoptosis was increased.Up-regulation of miR-365 inhibited proliferation,promoted apoptosis and decreased SIRT1 expression of LPS-treated nucleus pulposus cells,while down-regulation of miR-365 showed the opposite results.Up-regulation of SIRT1 could promote proliferation activity and inhibit apoptosis of PS-treated nucleus pulposus cells,while down-regulation of SIRT1 could obtain opposite results.Knockdown of SIRT1 could reverse the effects of miR-365 on proliferation and apoptosis of nucleus pulposus cells.MiR-365 negatively regulated SIRT1 expression.Conclusions MiR-365 regulates proliferation and apoptosis of nucleus pulposus cells through targeted regulation of SIRT1,providing a new tar

关 键 词：miR-365 细胞沉默信息调节因子1 髓核细胞 凋亡 增殖 

分 类 号：R681.5[医药卫生—骨科学] R392[医药卫生—外科学]