[1]李晓华,王 丹,晁雅静,等.类风湿关节炎滑膜组织中miR-150-5p表达对成纤维样滑膜细胞凋亡及炎症影响的实验研究[J].现代检验医学杂志,2026,41(02):88-94+99.[doi:10.3969/j.issn.1671-7414.2026.02.015]
　LI Xiaohua,WANG Dan,CHAO Yajing,et al.Experimental Study on the Impact of miR-150-5p Expression on Apoptosis and Inflammatory Responses in Fibroblast-Like Synoviocytes within Rheumatoid Arthritis Synovium[J].Journal of Modern Laboratory Medicine,2026,41(02):88-94+99.[doi:10.3969/j.issn.1671-7414.2026.02.015]
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类风湿关节炎滑膜组织中miR-150-5p表达对成纤维样滑膜细胞凋亡及炎症影响的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
88-94+99
栏目:
出版日期:
2026-03-15

文章信息/Info

Title:
Experimental Study on the Impact of miR-150-5p Expression on Apoptosis and Inflammatory Responses in Fibroblast-Like Synoviocytes within Rheumatoid Arthritis Synovium
文章编号:
1671-7414（2026）02-088-08
作者:
李晓华1a，王 丹2，晁雅静3，符亚璐1b
1.西安市人民医院（西安市第四医院） a.风湿病诊治中心；b.风湿免疫科，西安710004；2.西安市第五医院风湿免疫科，西安 710082；3.西安大兴医院风湿免疫科，西安 710003
Author(s):
LI Xiaohua1a, WANG Dan2, CHAO Yajing3, FU Yalu1b
1a. Rheumatology and Immunology Center; 1b. Department of Rheumatology and Immunology, Xi’an People’s Hospital (Xi’an Fourth Hospital), Xi’an 710004, China; 2. Department of Rheumatology and Immunology, Xi’an Fifth Hospital, Xi’an 710082, China; 3. Department of Rheumatology and Immunology, Xi’an Daxing Hospital, Xi’an 710003, China
关键词:
微小RNA-150-5p; 类风湿关节炎; 成纤维样滑膜细胞; 细胞凋亡; 炎症
分类号:
R593.22；R392.12
DOI:
10.3969/j.issn.1671-7414.2026.02.015
文献标志码:
A
摘要:
目的 探究类风湿关节炎(RA)滑膜组织中微小RNA(miR)-150-5p对RA-成纤维样滑膜细胞(FLS)凋亡与炎症的作用和机制。方法分别收集2021年1月~2023年1月在西安市人民医院(西安市第四医院)接受人工关节置换术的RA患者(RA组)和正常关节创伤患者(对照组)的滑膜组织各10例,采用实时荧光定量PCR(qRT-PCR)检测miR-150-5p表达水平。将RA-FLS细胞分为:①对照组、模型组、mimics-NC组和mimics组;②对照组、模型组、inhibitor-NC组和inhibitor组。其中模型组、mimics-NC组和mimics组、inhibitor-NC组和inhibitor组细胞培养液中均加入10ng/ml的肿瘤坏死因子-α(TNF-α)处理。转染完成后培养48h,采用qRT-PCR验证miR-150-5p转染效率,细胞计数试剂盒(CCK-8)检测RA-FLS细胞活力,5-Ethynyl-2’-脱氧尿苷(EdU)检测RA-FLS细胞增殖,流式细胞术检测RA-FLS细胞凋亡,酶联免疫吸附试验(ELISA)法检测RA-FLS细胞中白细胞介素(IL)-1、IL-6和TNF-α含量,蛋白质印迹(Westernblotting)检测Bcl-2相关X(Bax)蛋白、B细胞淋巴瘤/白血病-2(Bcl-2)基因、半胱天冬酶-3(CleavedCaspase-3)、Toll样受体5(TLR-5)、磷酸核因子κB(p-NF-κB)p65和核因子κB(NF-κB)p65蛋白表达水平。采用双荧光素酶报告基因实验和TargetScanHuman8.0数据库分析miR-150-5p与TLR-5的靶向结合关系。结果与对照组比较,RA组患者的滑膜组织中miR-150-5p(0.13±0.02vs1.00±0.02)的表达水平降低,差异具有统计学意义(t=52.829,P<0.05)。miR-150-5P过表达试验中,与对照组比较,模型组miR-150-5p、IL-10、Bax和Cleavedcasepase-3的水平以及细胞凋亡率均降低(t=16.520~47.522),IL-1β、IL-6、Bcl-2、TLR-5和p-NF-κBp65的水平及EdU阳性细胞率和细胞增殖活力均升高(t=14.038~33.839),差异具有统计学意义(均P<0.05)。与mimics-NC组比较,miR-150-5pmimics组miR-150-5p、IL-10、Bax和Cleavedcasepase-3的水平以及细胞凋亡率均升高(t=13.461~28.787),IL-1β、IL-6、Bcl-2、TLR-5、p-NF-κBp65水平及EdU阳性细胞率和细胞增殖活力均降低(t=11.670~28.146),差异具有统计学意义(均P<0.05)。miR-150-5P抑制表达实验中,与对照组比较,模型组miR-150-5p、IL-10、Bax和Cleavedcasepase-3的水平以及细胞凋亡率均降低(t=18.298~59.489),EdU阳性细胞率和细胞增殖活力及IL-1β、IL-6、Bcl-2、TLR-5和p-NF-κBp65水平均升高(t=15.623~28.497);与inhibitor-NC组比较,miR-150-5pinhibitor组miR-150-5p、IL-10、Bax和Cleavedcasepase-3的水平以及细胞凋亡率均降低(t=7.237~23.738),IL-1β、IL-6、Bcl-2、TLR-5和p-NF-κBp65水平及EdU阳性细胞率和细胞增殖活力均升高(t=3.071~6.252),差异具有统计学意义(均P<0.05)。miR-150-5p与TLR-5存在靶向结合位点。结论miR-150-5p与RA-FLS细胞增殖和凋亡调控密切相关,过表达miR-150-5p可以抑制RA-FLS细胞增殖和促炎因子的表达,并促进细胞凋亡,该作用机制可能与miR-150-5p调控TLR-5/NF-κBp65炎症信号通路有关。
Abstract:
Objective To investigate the effect and mechanism of microRNA (miR) -150-5p on apoptosis and inflammation of fibro-blast-like synoviocytes in rheumatoid arthritis synovium (RA-FLS). Methods Synovial tissues were collected from 10 patients with rheumatoid arthritis (RA group) and 10 patients with normal joint trauma (normal group) undergoing artificial joint replacement at Xi’an People’s Hospital (Xi’an Fourth Hospital) between January 2021 and January 2023. The expression level of miR-150-5p was detected by RT-qPCR method. RA-FLS cells were divided into control group, model group, mimics-NC group and mimics group; or control group, model group, inhibitor-NC group and inhibitor group. The model group, mimics-NC group, mimics group, inhibitor-NC group and inhibitor group were all treated with 10 ng/ml TNF-α. After 48h of transfection, the transfection ef-ficiency of miR-150-5p was validated by RT-qPCR. RA-FLS cell viability was detected by CCK-8 assay. RA-FLS cell proliferation was detected by EdU assay, and RA-FLS cell apoptosis was detected by flow cytometry. The levels of IL-1, IL-6 and TNF-α in RA-FLS cells were detected by ELISA, and the protein expression levels of Bax, Bcl-2, cleaved casepase-3, TLR-5, phospho-NF-κB p65 and NF-κB p65 were detected by Western blotting. The targeting relationship between miR-150-5p and TLR-5 was ana-lyzed by TargetScanHuman 8.0 database and dual luciferase reporter gene assay. Results Compared with the contorl group, the ex-pression level of miR-150-5p (0.13±0.02 vs 1.00±0.02) in synovial tissue of RA group was decreased, and the difference was statistically significant (t=52.829, P<0.05). In the miR-150-5p overepression exporiment, compared with the control group, the lev-els of miR-150-5p, IL-10, Bax and cleaved casepase-3 protein, as well as apoptosis rate in model group were decreased (t= 16.520~47.522). EdU positive cell rate and cell proliferation activity were increased, and the levels of IL-1β, IL-6 and Bcl-2, TLR-5, p-NF-κB p65 protein were increased (t=14.038~33.839), with statistically significant differences (all P<0.05). Compared with the mimics-NC group, the levels of miR-150-5p, IL-10, Bax and cleaved casepase-3 protein, along with apoptosis rate in miR-150-5p mimics group were increased (t=13.461~28.787), while EdU positive cell rate and cell proliferation activity along with the levels of IL-1β, IL-6 and Bcl-2, TLR-5, p-NF-κB p65 protein were decreased (t=11.670~28.146), with statistically significant differenc-es(all P<0.05). In the miR-150-5p inhibition exprossion experiment, compared with the control group, the model group showed de-creased levels of miR-150-5p, IL-10, Bax, and cleaved caspase-3, as well-as a reduced cell apoptosis rate (t=18.298~28.497 ), while the EdU-positive cell rate, cell proliferation activity, and the levels of IL-1β, IL-6, Bcl-2, ILR-5, and P-NF-κB p65 were increased (t=15.623~28.497); compared with inhibitor-NC group, the levels of miR-150-5p, IL-10, Bax, cleaved casepase-3 protein and apop-tosis rate in the inhibitor group were significantly decreased (t=7.237~23.738). Conversely, EdU positive cell rate, cell proliferation activity along with the levels of IL-1β, IL-6, Bcl-2, TLR-5, and p-NF-κB p65 protein levels were significantly increased (t=3.071~6.252), and the differences were statistically significant (all P<0.05). There are targeted binding sites between miR-150-5p and TLR-5. Conclusions miR-150-5p is closely related to the regulation of proliferation and apoptosis of RA-FLS cells. The over-expression of miR-150-5p enhances apoptosis in RA-FLS cells while inhibiting cell proliferation and pro-inflammatory factor ex-pression, potentially through its regulation of the TLR-5/NF-κB p65 inflammatory signaling pathway.

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备注/Memo

备注/Memo:
基金项目：西安市卫生健康委员会科研项目（2024yb11）。
作者简介：李晓华（1989-），女，硕士研究生，主治医师，研究方向：风湿免疫性疾病，E-mail：XiaoHuaL4@126.com。
通讯作者：符亚璐（1988-），女，硕士研究生，主治医师，研究方向：弥漫性结缔组织病的诊断与治疗，E-mail：fu_yalu@126.com。
更新日期/Last Update: 2026-03-15