中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (12): 2484-2491.doi: 10.12307/2025.388

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

刺芒柄花素对白细胞介素1β诱导软骨细胞损伤影响的机制

单继新，叶锐彬，巨少华，王  强   

  1. 成都体育学院附属体育医院正骨科，四川省成都市  610041
  • 收稿日期:2024-03-22 接受日期:2024-06-28 出版日期:2025-04-28 发布日期:2024-09-10
  • 通讯作者: 叶锐彬，主任医师，成都体育学院附属体育医院正骨科，四川省成都市 610041
  • 作者简介:单继新，男，1986年生，山东省泗水市人，汉族，硕士，主治医师，主要从事中医骨伤研究。
  • 基金资助:
    四川省中医药管理局科学技术研究专项课题项目(2023MS271)，项目负责人：王强；四川省自然科学基金项目(2023NSFC1803)，项目负责人：巨少华

Mechanism underlying the effect of formononetin on interleukin-1beta-induced chondrocyte injury #br#
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Shan Jixin, Ye Ruibin, Ju Shaohua, Wang Qiang   

  1. Department of Bonesetting, Affiliated Sport Hospital of Chengdu Sport University, Chengdou 610041, Sichuan Province, China
  • Received:2024-03-22 Accepted:2024-06-28 Online:2025-04-28 Published:2024-09-10
  • Contact: Ye Ruibin, Chief physician, Department of Bonesetting, Affiliated Sport Hospital of Chengdu Sport University, Chengdou 610041, Sichuan Province, China
  • About author:Shan Jixin, Master, Attending physician, Department of Bonesetting, Affiliated Sport Hospital of Chengdu Sport University, Chengdou 610041, Sichuan Province, China
  • Supported by:
    Sichuan Provincial Administration of Traditional Chinese Medicine Scientific and Technological Research Special Project, No. 2023MS271 (to WQ); Sichuan Provincial Natural Science Foundation, No. 2023NSFC1803 (to JSH)

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摘要：




文题释义：
刺芒柄花素：为异黄酮类化合物，别称刺芒柄花素、芒柄花黄素，主要存在于甘草、黄芪、红车轴草、葛根等豆科植物中，也可化学合成，可保护植物免受环境胁迫或病害，具有抗氧化、抗炎、抗细胞凋亡等生物学作用，在预防前列腺癌、乳腺癌、结肠癌等肿瘤以及改善骨质疏松方面有一定作用。
微小RNA：为内源性非编码小RNA，可通过切割、抑制翻译调控靶基因的表达水平，进而影响细胞的生物学过程，具有抗炎、抗氧化、调控细胞增殖、凋亡等多种生物学功能，与骨关节炎、癌症、神经系统疾病等的发生发展密切相关。

背景：刺芒柄花素是一种异黄酮类化合物，广泛存在于红车轴草、黄芪、鸡血藤中，具有抑制氧化应激、炎症因子释放及细胞凋亡的作用。
目的：探讨刺芒柄花素对白细胞介素1β诱导软骨细胞损伤的影响及机制。
方法：①收集骨关节炎患者及单纯半月板损伤患者的软骨组织，采用实时定量PCR检测miR-135b-5p表达。②体外培养人软骨细胞，分9组培养：正常对照组不进行任何处理，培养48 h；白细胞介素1β组加入白细胞介素1β处理48 h；白细胞介素1β+刺芒柄花素低浓度组加入25 μmol/L刺芒柄花素处理24 h后加入白细胞介素1β处理48 h；白细胞介素1β+刺芒柄花素中浓度组加入50 μmol/L刺芒柄花素处理24 h后加入白细胞介素1β处理48 h；白细胞介素1β+刺芒柄花素高浓度组加入100 μmol/L刺芒柄花素处理24 h后加入白细胞介素1β处理48 h；白细胞介素1β+miR NC组转染miR NC 6 h后加入白细胞介素1β处理48 h；白细胞介素1β+miR-135b-5p组转染miR-135b-5p模拟物6 h后加入白细胞介素1β处理48 h；白细胞介素1β+刺芒柄花素高浓度+anti-miR-NC组转染anti-miR-NC 6 h后加入100 μmol/L刺芒柄花素处理24 h，再加入白细胞介素1β处理48 h；白细胞介素1β+刺芒柄花素高浓度+anti-miR-135b-5p组转染anti-miR-135b-5p 6 h后加入100 μmol/L刺芒柄花素处理24 h，再加入白细胞介素1β处理48 h。处理结束后进行相关检测。
结果与结论：①骨关节炎患者软骨组织中miR-135b-5p表达低于单纯半月板损伤患者(P < 0.05)。②与正常对照组比较，白细胞介素1β组软骨细胞miR-135b-5p表达、增殖活力、超氧化物歧化酶活性、Ⅱ型胶原蛋白、Bcl-2蛋白表达均降低(P < 0.05)，细胞凋亡率、乳酸脱氢酶活性、丙二醛水平、促炎因子水平、基质金属蛋白酶13蛋白、Bax蛋白表达均升高(P < 0.05)；刺芒柄花素可抑制白细胞介素1β对软骨细胞造成的损伤，并且呈现浓度依赖性。转染miR-135b-5p模拟物可提升白细胞介素1β组miR-135b-5p表达，抑制白细胞介素1β对软骨细胞造成的损伤；转染anti-miR-135b-5p可降低白细胞介素1β+刺芒柄花素高浓度组miR-135b-5p表达，抑制刺芒柄花素发挥软骨细胞作用。③结果表明，刺芒柄花素对白细胞介素1β诱导软骨细胞损伤的保护作用可能与调控miR-135b-5p表达有关。
https://orcid.org/0009-0009-7922-1319(单继新)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 刺芒柄花素, miR-135b-5p, 软骨细胞, 氧化应激, 炎症, 细胞增殖, 细胞凋亡

Abstract: BACKGROUND: Formononetin is an isoflavonoid compound widely found in red clover, astragalus, and chickweed, which has the ability to inhibit oxidative stress, inflammatory factor release, and apoptosis. 
OBJECTIVE: To investigate the effect of formononetin on interleukin-1β-induced chondrocyte injury and its mechanism. 
METHODS: (1) Cartilage tissues from patients with osteoarthritis and patients with simple meniscus injury were collected, and real-time quantitative PCR was used to detect miR-135b-5p expression. (2) Human chondrocytes were cultured in vitro, and then divided them into nine groups: cells in normal control group were cultured for 48 hours with no treatment; cells in interleukin-1β group were treated with interleukin-1β for 48 hours; cells in interleukin-1β+low-dose formononetin group were treated with 25 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+middle-dose formononetin group were treated with 50 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+high-dose formononetin group were treated with 100 μmol/L formononetin for 24 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+miR NC group were treated with miR NC for 6 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+miR-135b-5p group were treated with miR-135b-5p mimics for 6 hours followed by treatment with interleukin-1β for 48 hours; cells in interleukin-1β+high-dose formononetin+anti-miR-NC group were treated with anti-miR-NC for 6 hours, then treated with 100 μmol/L formononetin for 24 hours, and finally treated with interleukin-1β for 48 hours; cells in interleukin-1β+high-dose formononetin+anti-miR-135b-5p group were treated with anti-miR-135b-5p for 6 hours, then treated with 100 μmol/L formononetin for 24 hours, and finally treated with interleukin-1β for 48 hours. Relevant tests are performed after treatment.
RESULTS AND CONCLUSION: The expression level of miR-135b-5p in cartilage tissue of patients with osteoarthritis was significantly lower than that of patients with simple meniscus injury (P < 0.05). Compared with the normal control group, the expression level of miR-135b-5p, proliferative ability, activity of superoxide dismutase, and expression levels of collagenase II protein and Bcl-2 protein in chondrocytes were lower in the interleukin-1β group (P < 0.05), while apoptotic rate, lactate dehydrogenase activity, malondialdehyde level, levels of proinflammatory factors, and expression levels of matrix metalloproteinase-13 protein and Bax protein were higher in the interleukin-1β group (P < 0.05). Formononetin inhibited chondrocyte damage caused by interleukin-1β in a concentration-dependent manner. Transfection of miR-135b-5p mimics elevated miR-135b-5p expression in the interleukin-1β group and inhibited chondrocyte damage induced by interleukin-1β; transfection of anti-miR-135b-5p decreased miR-135b-5p expression in the interleukin-1β+high-dose formononetin group and inhibited the effect of formononetin on chondrocytes. To conclude, the protective effect of formononetin on chondrocyte injury induced by interleukin-1β may be related to the regulation of miR-135b-5p expression.

Key words: formononetin, miR-135b-5p, chondrocyte, oxidative stress, inflammation, proliferation, apoptosis

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