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| LncRNA FGD5-AS1靶向miR-103a-3p对IL-1β诱导的关节软骨细胞凋亡的机制研究 |
| Mechanism of LncRNA FGD5-AS1 on IL-1β-induced articular chondrocyte apoptosis through targeting regulation of miR-103a-3p expression |
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| DOI：10.3969/j.issn.1006-7108.2020.06.009 |
| 中文关键词: LncRNA FGD5-AS1 miR-103a-3p IL-1β 关节软骨细胞 凋亡 |
| 英文关键词:LncRNA FGD5-AS1 miR-103a-3p IL-1β articular chondrocytes apoptosis |
| 基金项目:内蒙古自治区高等学校科学研究项目（NJZZ20130） |
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| 中文摘要: |
| 目的 探讨LncRNA FGD5-AS1靶向miR-103a-3p对IL-1β诱导的关节软骨细胞损伤及细胞凋亡的影响与分子机制。方法 体外培养大鼠关节软骨细胞，分为NC组、IL-1β组（IL-1β浓度为 10 ng/mL）。采用qRT-PCR检测细胞中FGD5-AS1、miR-103a-3p的表达水平。分别将pcDNA-FGD5-AS1、miR-103a-3p mimics转染至软骨细胞，随后使用IL-1β处理48 h。采用ELISA检测IL-6、TNF-α、IL-8水平；流式细胞术检测细胞凋亡率；双荧光素酶报告实验验证FGD5-AS1与miR-103a-3p的靶向关系；Western blot检测Bax、Cyt C、Cleaved Caspase-3、p-NF-κB p65、p-IκBα蛋白表达量。结果 与NC组相比，IL-1β组软骨细胞中FGD5-AS1的表达水平显著降低（P＜0.05），miR-103a-3p、Bax、Cyt C、Cleaved Caspase-3的表达水平显著升高（P＜0.05），IL-6、TNF-α、IL-8水平显著升高（P＜0.05），细胞凋亡率显著升高（P＜0.05）；FGD5-AS1过表达后可明显降低IL-6、TNF-α、IL-8、p-NF-κB p65、p-IκBα水平（P＜0.05），降低细胞凋亡率（P＜0.05），抑制Bax、Cyt C、Cleaved Caspase-3表达（P＜0.05）；双荧光素酶报告实验证实FGD5-AS1靶向结合miR-103a-3p并可负向调控miR-103a-3p的表达（P＜0.05）；miR-103a-3p过表达可明显逆转FGD5-AS1过表达对细胞凋亡及炎症反应的抑制作用（P＜0.05）。结论 LncRNA FGD5-AS1可通过靶向负调控miR-103a-3p的表达从而抑制IL-1β诱导的关节软骨细胞炎症反应及细胞凋亡，其作用机制可能通过抑制NF-κB信号通路激活有关。 |
| 英文摘要: |
| ObjectiveTo investigate the effect of LncRNA FGD5-AS1 targeting miR-103a-3p on IL-1β-induced articular chondrocyte injury and apoptosis and its molecular mechanism. Methods Rat articular chondrocytes were cultured in vitro and divided into NC group and IL-1β group (IL-1β 10 ng / mL). The expression levels of FGD5-AS1 and miR-103a-3p in cells were detected by qRT-PCR. PcDNA-FGD5-AS1, miR-103a-3p mimics were transfected into chondrocytes respectively, and then treated with IL-1β for 48 h. ELISA was used to detect the levels of IL-6, TNF-α and IL-8. Flow cytometry was used to detect the apoptotic rate. The dual luciferase report experiment verified the targeting relationship between FGD5-AS1 and miR-103a-3p. Western blot was used to detect the expression levels of Bax, Cyt C, Cleaved Caspase-3, p-NF-κB p65, and p-IκBα protein . Results Compared with the NC group, the expression level of FGD5-AS1 in chondrocytes in the IL-1β group was significantly reduced (P<0.05), and the expression levels of miR-103a-3p, Bax, Cyt C, and Cleaved Caspase-3 were significantly increased (P <0.05), the levels of IL-6, TNF-α and IL-8 were significantly increased (P <0.05), and the apoptosis rate was significantly increased (P<0.05). FGD5-AS1 overexpression could significantly reduce the levels of IL-6, TNF-α, IL-8, p-NF-κB p65, and p-IκBα (P <0.05), reduced the rate of apoptosis (P<0.05), and inhibited Bax, Cyt C, and Cleaved Caspase-3 expression (P<0.05). The double luciferase reporting experiment confirmed that FGD5-AS1 targets miR-103a-3p and could negatively regulate the expression of miR-103a-3p (P<0.05). Overexpression of miR-103a-3p could significantly reverse the inhibitory effect of FGD5-AS1 overexpression on apoptosis and inflammatory response (P<0.05). Conclusion LncRNA FGD5-AS1 could negatively regulate the expression of miR-103a-3p, thereby inhibiting IL-1β-induced articular chondrocyte inflammation and apoptosis, and its mechanism may be related to the inhibition of NF-κB signaling pathway activation. |
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