橙皮素通过circ_0039411/miR-296-5p影响IL-1β诱导的软骨细胞凋亡  

Hesperetin affects IL-1β-induced chondrocyte apoptosis through circ_0039411/miR-296-5p

作　　者：郭玲[1] 王兆领[1] 黄振[1] GUO Ling;WANG Zhaoling;HUANG Zhen(Department of Pharmacy,Tengzhou Central People’s Hospital,Tengzhou 277599,China)

机构地区：[1]滕州市中心人民医院药学部,277599

出　　处：《免疫学杂志》2022年第2期151-156,163,共7页Immunological Journal

摘　　要：目的探讨橙皮素对IL-1β诱导的软骨细胞损伤的影响及分子机制。方法关节软骨细胞分为对照(control)组、IL-1β组、IL-1β+橙皮素低、中、高剂量组、IL-1β+si-circ_0039411组、IL-1β+si-NC组、IL-1β+HES+pcDNA-circ_0039411组、IL-1β+HES+pcDNA组。酶联免疫吸附试验(ELISA)检测IL-6、TNF-α、IFN-γ水平;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达;实时荧光定量PCR(RT-qPCR)检测circ_0039411和miR-296-5p的表达水平;双荧光素酶报告实验检测circ_0039411和miR-296-5p的靶向关系。结果橙皮素低、中、高浓度组软骨细胞中IL-6、TNF-α、IFN-γ水平降低,cleaved-caspase3和cleaved-caspase9表达水平降低,软骨细胞凋亡率降低,circ_0039411表达水平降低,miR-296-5p表达水平升高,且呈浓度依赖性(P<0.05)。circ_0039411靶向调控miR-296-5p;抑制circ_0039411表达后,IL-6、TNF-α、IFN-γ水平降低,cleaved-caspase3和cleaved-caspase9表达水平降低,软骨细胞凋亡率降低(P<0.05)。circ_0039411过表达逆转了橙皮素对IL-1β诱导的软骨细胞损伤的作用。结论橙皮素可能通过调控circ_0039411/miR-296-5p抑制IL-1β诱导的软骨细胞损伤。This study was performed to investigate the effect of hesperetin on IL-1β-induced chondrocyte damage and its molecular mechanism.Articular chondrocytes were divided into control group,IL-1βgroup,IL-1β+hesperetin low,medium and high dose groups,IL-1β+si-circ_0039411 group,IL-1β+si-NC group,IL-1β+HES+pcDNA-circ_0039411 group,IL-1β+HES+pcDNA group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of IL-6,TNF-α,IFN-γ;flow cytometry was applied to detect cell apoptosis;Western blot was used to detect protein expression;real-time fluorescent quantitative PCR(RT-qPCR)was utilized to detect the expression levels of circ_0039411 and miR-296-5p;dual luciferase reporter experiment was performed to detect the targeting relationship between circ_0039411 and miR-296-5p.Data showed that the levels of IL-6,TNF-αand IFN-γin chondrocytes of the hesperetin low,medium and high groups were decreased,the expression levels of cleaved-caspase3 and cleaved-caspase9 were decreased,the apoptosis rate of chondrocytes was decreased,the expression level of circ_0039411 was decreased,and the expression level of miR-296-5p was increased in concentration-dependent manner(P<0.05).circ_0039411 targeted and regulated miR-296-5p.After inhibiting the expression of circ_0039411,the levels of IL-6,TNF-α,IFN-γwere decreased,the expression levels of cleavedcaspase3 and cleaved-caspase9 were decreased,and the apoptosis rate of chondrocytes was decreased(P<0.05).The overexpression of circ_0039411 could reversed the effect of hesperetin on the chondrocyte damage induced by IL-1β.In conclusion,hesperetin may alleviate IL-1β-induced chondrocyte damage by regulating circ_0039411/miR-296-5p.

关 键 词：橙皮素 circ_0039411 miR-296-5p 软骨细胞 损伤 

分 类 号：R684.3[医药卫生—骨科学]