中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (28): 4565-4571.doi: 10.12307/2024.463

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

长基因间非蛋白编码RNA 00707对骨关节炎软骨细胞损伤及炎症因子分泌的影响

褚  凯，孙建华   

  1. 石河子大学医学院第一附属医院骨科中心，新疆维吾尔自治区石河子市  832000
  • 收稿日期:2022-11-10 接受日期:2023-09-02 出版日期:2024-10-08 发布日期:2023-11-27
  • 通讯作者: 孙建华，博士，主任医师，石河子大学医学院第一附属医院骨科中心，新疆维吾尔自治区石河子市 832000
  • 作者简介:褚凯，男，1992年生，安徽省亳州市人，硕士，主要从事骨外科疾病研究。
  • 基金资助:


Effects of long intergenic non-protein coding RNA 00707 on chondrocyte injury and inflammatory factor secretion in osteoarthritis

Chu Kai, Sun Jianhua   

  1. Orthopedic Center, The First Affiliated Hospital of Shihezi University School of Medicine, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
  • Received:2022-11-10 Accepted:2023-09-02 Online:2024-10-08 Published:2023-11-27
  • Contact: Sun Jianhua, MD, Chief physician, Orthopedic Center, The First Affiliated Hospital of Shihezi University School of Medicine, Shihezi 832000, Xinjiang Uygur Autonomous Region, China
  • About author:Chu Kai, Master, Orthopedic Center, The First Affiliated Hospital of Shihezi University School of Medicine, Shihezi 832000, Xinjiang Uygur Autonomous Region, China

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摘要：


文题释义：

骨关节炎：为一种退行性关节疾病，是导致老年人残疾和预期寿命缩短的主要原因之一，临床治疗效果欠佳。软骨细胞的异常凋亡和炎症反应对骨关节炎的发生发展至关重要，调控软骨细胞凋亡和/或抑制炎症反应可能是骨关节炎新的治疗方向。  
软骨细胞损伤模型：白细胞介素1β是一种促炎因子，在骨关节炎发生发展过程中起到关键作用。白细胞介素1β不仅可诱导骨关节炎软骨细胞产生炎症递质，同时还可诱导软骨细胞降解代谢，促进大量含有透明质酸的小片段生成，使得炎症级联扩大，因此通常采用白细胞介素1β处理软骨细胞来模拟骨关节炎的炎症状态。


背景：长基因间非蛋白编码RNA(long intergenic non-protein coding RNA，LINC RNA) 00707和miR-423-5p均参与了骨关节炎的发生发展过程，Starbase预测LINC00707和miR-423-5p存在互补序列，但二者是否存在相互作用仍需要进一步研究。

目的：考察LINC0070是否通过靶向miR-423-5p影响骨关节炎的软骨细胞损伤及炎症因子分泌。
方法：将关节软骨细胞分8组培养：①空白对照组不进行任何处理；②IL-1β组用含10 ng/mL白细胞介素1β的培养基处理48 h；③IL-1β+si-NC组将si-NC转染至细胞中，然后用白细胞介素1β处理48 h；④IL-1β+si-LINC00707组将si-LINC00707染至细胞中，然后用白细胞介素1β处理48 h；⑤IL-1β+miR-NC组将miR-NC转染至细胞中，然后用白细胞介素1β处理48 h；⑥IL-1β+miR-423-5p组将miR-423-5p模拟物转染至细胞中，然后用白细胞介素1β处理48 h；⑦IL-1β+si-LINC00707+anti-miR-NC组将si-LINC00707与anti-miR-NC共转染至细胞中，然后用白细胞介素1β处理48 h；⑧IL-1β+si-LINC00707+anti-miR-423-5p组将si-LINC00707与anti-miR-423-5p共转染至细胞中，然后用白细胞介素1β处理48 h。干预结束后进行相关检测。

结果与结论：①与空白对照组相比，IL-1β组软骨细胞中LINC00707基因表达量增加、miR-423-5p基因表达量减少，细胞凋亡率及裂解caspase3、裂解caspase9蛋白表达量增加，培养液内肿瘤坏死因子α、白细胞介素6水平升高，白细胞介素10水平降低(P均< 0.05)；与IL-1β组相比，IL-1β+si-LINC00707组软骨细胞中LINC00707基因表达量减少、miR-423-5p基因表达量增加，细胞凋亡率及裂解caspase3、裂解caspase9蛋白表达量减少，培养液内肿瘤坏死因子α、白细胞介素6水平降低，白细胞介素10水平升高(P均< 0.05)。②与IL-1β+miR-NC组相比，IL-1β+miR-423-5p组软骨细胞中miR-423-5p基因表达量增加，细胞凋亡率及裂解caspase3、裂解caspase9蛋白表达量减少，培养液内肿瘤坏死因子α、白细胞介素6水平降低，白细胞介素10水平升高(P均< 0.05)。③与IL-1β+si-LINC00707+anti-miR-NC组相比，IL-1β+si-LINC00707+anti-miR-423-5p组软骨细胞中miR-423-5p基因表达量减少，细胞凋亡率及裂解caspase3、裂解caspase9蛋白表达量增加，培养液内肿瘤坏死因子α、白细胞介素6水平升高，白细胞介素10水平降低(P均< 0.05)。④结果表明，抑制LINC00707可靶向miR-423-5p减轻白细胞介素1β诱导的关节软骨细胞凋亡和炎症反应。

https://orcid.org/0009-0008-5179-8290(褚凯)

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: 骨关节炎, LINC00707, miR-423-5p, 白细胞介素1β, 软骨细胞, 细胞凋亡, 炎性因子

Abstract: BACKGROUND: Long intergenic non-protein coding RNA 00707 (LINC00707) and microRNA-423-5p (miR-423-5p) are both involved in the occurrence and development of osteoarthritis. Starbase predicts that LINC00707 and miR-423-5p have complementary sequences, but whether LINC00707 and miR-423-5p interact to regulate the progress of osteoarthritis still needs further research.
OBJECTIVE: To investigate whether LINC00707 targets miR-423-5p to affect chondrocyte injury and inflammatory factor secretion in osteoarthritis. 
METHODS: Articular chondrocytes were divided into eight groups: (1) blank control group was given no treatment; (2) interleukin (IL)-1β group was cultured with 10 ng/mL IL-1β for 48 hours; (3) IL-1β+si-NC group was transfected with si-NC and then treated with 10 ng/mL IL-1β for 48 hours; (4) IL-1β+si-LINC00707 group was transfected with si-LINC00707 and then treated with 10 ng/mL IL-1β for 48 hours; (5) IL-1β+miR-NC group was transfected with miR-NC and then treated with 10 ng/mL IL-1β for 48 hours; (6) IL-1β+miR-423-5p group was transfected with miR-423-5p mimic and then treated with 10 ng/mL IL-1β for 48 hours; (7) IL-1β+si-LINC00707+anti-miR-NC group was co-transfected with si-LINC00707 and anti-miR-NC and then treated with 10 ng/mL IL-1β for 48 hours; (8) IL-1β+si-LINC00707+anti-miR-423-5p group was co-transfected with si-LINC00707 and anti-miR-423-5p and then treated with 10 ng/mL IL-1β for 48 hours. Relevant tests were performed at the end of the intervention.
RESULTS AND CONCLUSION: Compared with the blank control group, the mRNA expression of LINC00707, apoptosis rate, protein expression of C-caspase3 and C-caspase9, and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were increased in the IL-1β group, while there was a decrease in miR-423-5p expression and IL-10 level (P < 0.05). Compared with the IL-1β group, the mRNA expression of LINC00707, apoptosis rate, protein expression of C-caspase3 and C-caspase9, and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were decreased in the IL-1β+si-LINC00707 group, while miR-423-5p expression and IL-10 level increased (P < 0.05). Compared with the IL-1β+miR-NC group, the protein expression of C-caspase3 and C-caspase9 and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were decreased in the IL-1β+miR-423-5p group, while miR-423-5p expression and IL-10 level increased (P < 0.05). Compared with the IL-1β+si-LINC00707+anti-miR-NC group, apoptosis rate, protein expression of C-caspase3 and C-caspase9, and levels of tumor necrosis factor-α and IL-6 in articular chondrocytes were increased in the IL-1β+si-LINC00707+anti-miR-423-5p group, while miR-423-5p expression and IL-10 level decreased (P < 0.05). To conclude, inhibiting LINC00707 by targeting miR-423-5p can reduce IL-1β-induced apoptosis and inflammation in articular chondrocytes.

Key words: osteoarthritis, LINC00707, miR-423-5p, interleukin-1β, chondrocyte, apoptosis, inflammatory factor

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