[1]何　至,颜端国,严　林.人退变椎间盘髓核组织中 miR-146a的表达及对 NP细胞增殖、凋亡和炎性细胞因子的影响作用机制[J].现代检验医学杂志,2022,37(01):72-76.[doi:10.3969/j.issn.1671-7414.2022.01.015]
　HE Zhi,YAN Duan-guo,YAN Lin.Expression of miR-146a in the Nucleus Pulposus Tissue of Human Degenerative Intervertebral Disc and Its Effect Mechanism onthe Proliferation, Apoptosis and Inflammatory Cytokines of NP Cells[J].Journal of Modern Laboratory Medicine,2022,37(01):72-76.[doi:10.3969/j.issn.1671-7414.2022.01.015]
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人退变椎间盘髓核组织中 miR-146a的表达及对 NP细胞增殖、凋亡和炎性细胞因子的影响作用机制()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年01期
页码:
72-76
栏目:
论　著
出版日期:
2022-01-15

文章信息/Info

Title:
Expression of miR-146a in the Nucleus Pulposus Tissue of Human Degenerative Intervertebral Disc and Its Effect Mechanism onthe Proliferation, Apoptosis and Inflammatory Cytokines of NP Cells
文章编号:
1671-7414（2022）01-072-06
作者:
何　至1; 颜端国1; 严　林2
（1. 湖北省监利市人民医院骨外科，湖北监利433300；2. 华中科技大学同济医学院附属荆州医院骨外科，湖北荆州434020）
Author(s):
HE Zhi1;  YAN Duan-guo1;  YAN Lin2
（1.Department of Orthopeadic Surgery, Jianli People’s Hospital of HubeiProvince, Hubei Jianli 433300, China; 2.Department of Orthopeadic Surgery, Jingzhou Hospital Affiliated to TongjiMedical College of Huazhong University of Science and Technology, Hubei Jingzhou 434020, China）
关键词:
椎间盘退变; 微小 RNA146a; Toll样受体 4; 炎性反应; 增殖; 凋亡
分类号:
R681.53；R392.11
DOI:
10.3969/j.issn.1671-7414.2022.01.015
文献标志码:
A
摘要:
目的　检测人退变椎间盘髓核组织中 miR-146a的表达变化，并探讨其可能的作用机制。方法　实时荧光定量 PCR (qRT-PCR) 检测人退变腰椎间盘髓核组织中 miR-146a表达。用脂多糖（LPS）刺激人髓核（nucleus pulposus，NP）细胞模拟椎间盘变性，qRT-PCR检测 NP细胞中 miR-146a的表达变化。将 miR-146a mimics或 miR-146a inhibitor转染至 NP细胞，然后 LPS刺激 NP细胞，观察 miR-146a对 NP细胞增殖活性、凋亡、 TLR4蛋白表达及 IL-1β,TNF-α和 IL-6等炎性因子水平的影响。结果　miR-146a在椎间盘退变（intervertebral disc degeneration, IVDD组）椎间盘髓核组织中表达（0.65±0.12）明显低于对照组（ 1.01±0.10），miR-146a在 LPS组 NP细胞中表达（ 0.29±0.11）明显低于阴性对照组（1.06±0.07），差异均有统计学意义（ t=11.856，10.229，均 P＜ 0.01）。与阴性对照组比较， LPS组上清液中的 IL-1β, TNF-α和 IL-6含量明显升高，差异均有统计学意义（ t=15.572~23.354，均 P＜ 0.001）。上调 NP细胞 miR-146a表达，能够提高 LPS刺激的 NP细胞的增殖活性（ t=4.436~7.995, 均 P＜ 0.05），降低其凋亡率（ t=9.255，P=0.001），降低 TLR4蛋白（ t=5.881，P=0.004）和 IL-1β，TNF-α和 IL-6等炎性细胞因子水平（ t=8.854~10.772，均 P＜ 0.01）；而下调 NP细胞 miR-146a表达则出现相反的结果（ t=2.977~6.777，均 P<0.05）。结论　miR-146a在退变椎间盘髓核组织中表达降低， miR-146a可能能够通过靶向 TLR4调控 NP细胞的增殖、凋亡和炎症反应，为椎间盘退变的生物治疗提供了新的方向。
Abstract:
Objective　To detect the expression of miR-146a in nucleus pulposus of human degenerative intervertebral disc andexplore its possible mechanism. Methods　qRT-PCR was used to detect the expression of miR-146a in human degenerativelumbar disc nucleus pulposus.Human nucleus pulposus (NP) cells were stimulated with lipopolysaccharide (LPS) to simulateintervertebral disc degeneration. The expression of miR-146a in NP cells was detected by qRT-PCR.miR-146a mimics or miR-146a inhibitor were transfected into NP cells, and then LPS stimulated NP cells. The effects of miR-146a on proliferation,apoptosis, TLR4 protein expression and expression of IL-1β, TNF-α and IL-6 were observed. Results　 The expression ofmiR-146a in the intervertebral disc nucleus pulposus tissue of the (intervertebral disc degeneration IVDD) group (0.65±0.12)was significantly lower than that of the control group (1.01±0.10), and the expression of miR-146a in the NP cells of the LPSgroup (0.29±0.11) was significantly lower than that of the negative control group (1.06±0.07), the difference was statisticallysignificant (t=11.856, 10.229, all P<0.01). Compared with the negative control group, the levels of IL-1β, TNF-α and IL-6 inthe supernatant of the LPS group were significantly increased, and the differences were statistically significant (t=15.572~23.354,all P<0.001) .Up regulation of miR-146a expression in NP cells could increase the proliferation activity (t=4.436~7.995, allP<0.05), reduce the apoptosis rate (t=9.255，P=0.001), and decreased the levels of TLR4 protein (t=5.881，P=0.004) andinflammatory cytokines such as IL-1β, TNF-α and IL-6 (t=8.854~10.772，all P ＜ 0.01), while down regulating the expressionof miR-146a in NP cells had the opposite effect (t=2.977~6.777, all P<0.05). Conclusion　The expression of miR-146a innucleus pulposus of degenerative intervertebral disc was decreased. MiR-146a may be able to regulate the proliferation, apoptosisand inflammatory reaction of NP cells by targeting TLR4, which provides a new direction for the biological treatment ofintervertebral disc degeneration.

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备注/Memo

备注/Memo:
作者简介：何至（1982-），男，本科，主治医师，研究方向：脊柱外科疾病，E-mail：hezhigwk@126.com。
通讯作者：严林（1983-），男，硕士，副主任医师，研究方向：脊柱外科基础及临床研究，E-mail: sg4206226@126.com。
更新日期/Last Update: 1900-01-01