积雪草苷调控miR-342-5p/AQP3轴保护IL-1β诱导的软骨细胞损伤  

Asiaticoside Modulates MIR-342-5p/AQP3 Axis Protection Against IL-1-Induced Chondrocyte Injury

作　　者：付涟桥[1] 郭焱雄[1] 李坚[1] 周新林[1] FU Lianqiao;GUO Yanxiong;LI Jian;ZHOU Xinlin(Department of Emergency,South Hospital of Chenzhou First People’s Hospital,Chenzhou 423000,China)

机构地区：[1]郴州市第一人民医院南院急诊科,郴州423000

出　　处：《实验动物科学》2020年第3期10-18,共9页Laboratory Animal Science

摘　　要：目的探讨积雪草苷(Ass)对白细胞介素1β(IL-1β)诱导的软骨细胞损伤的影响及作用机制。方法不同浓度的Ass作用IL-1β诱导软骨细胞24 h,流式细胞仪检测细胞凋亡,酶联免疫吸附法检测细胞培养上清液中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平,实时荧光定量PCR检测细胞中miR-342-5p和水通道蛋白3(AQP3)mRNA表达水平,Western Blot法检测B淋巴细胞瘤-2(Bcl-2)、B淋巴细胞瘤-2相关蛋白(Bax)和AQP3蛋白表达水平。双荧光素酶报告基因实验验证miR-342-5p与AQP3靶向关系。转染miR-342-5p抑制剂或AQP3过表达载体至软骨细胞,上述相同方法检测抑制miR-342-5p表达或AQP3过表达对IL-1β诱导的软骨细胞凋亡及IL-6和TNF-α表达的影响。结果Ass可抑制IL-1β诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达及miR-342-5p表达(P<0.05),促进Bcl-2蛋白、AQP3 mRNA和蛋白表达(P<0.05)。miR-342-5p在软骨细胞中负调控AQP3表达。抑制miR-342-5p或AQP3过表达后,IL-1β诱导的软骨细胞凋亡率、Bax蛋白、IL-6和TNF-α表达降低(P<0.05),Bcl-2蛋白表达升高(P<0.05)。抑制AQP3表达逆转了抑制miR-342-5p对IL-1β诱导的软骨细胞凋亡、Bax和Bcl-2蛋白及IL-6和TNF-α表达的影响。结论Ass可抑制IL-1β诱导软骨细胞凋亡和炎症反应,其可能通过调控miR-342-5p/AQP3保护软骨细胞损伤。Objective To investigate the effect and mechanism of asiaticoside(Ass)on chondrocyte injury induced by interleukin 1β(IL-1β).Method After different concentrations of Ass were applied to(IL-1)Ass on chondrocyte injury induced by interleukin 1β(IL-1β).echaniccellsapoptosis,enzyme-linked immunosorbent assay was used to detect the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cell culture supernatants,real-time quantitative PCR was used to detect the expression levels of miR-342-5 p and aquaporin 3(AQP3)mRNA,and Western Blot method was used to detect the expression levels of B lymphoma-2(Bcl-2),B lymphoma-2 related protein(Bax)and AQP3 protein.The double luciferase reporter experiment verified the targeting relationship between miR-342-5 p and AQP3.After miR-342-5 p inhibitor or AQP3 over expression vector was transfected into chondrocytes,the same method described above were used to detect the effect of inhibiting miR-342-5 porover expressing AQP3 on IL-1β-induced chondrocyte apoptosis and the expression of IL-6 and TNF-α.Result Ass could inhibit the apoptosis rate of chondrocytes induced by IL-1β and the expression of Bax protein,IL-6 and TNF-α and miR-342-5 p(P<0.05),and promote the expression of Bcl-2 protein,AQP3 mRNA and protein(P<0.05).miR-342-5 p negatively regulated AQP3 expression in chondrocytes.After inhibiting miR-342-5 p or over expressing AQP3,the apoptosis rate of chondrocytes induced by IL-1β and the expression of Bax protein,IL-6 and TNF-α were decreased(P<0.05),while the expression of Bcl-2 protein was increased(P<0.05).Inhibiting AQP3 reversed the effect of inhibiting miR-342-5 p on IL-1β-induced chondrocyte apoptosis and the expression of Bax,Bcl-2,IL-6 and TNF-α.Conclusion Ass can inhibit the apoptosis and inflammatory response of chondrocytes induced by IL-1 rocyte apoptosis and the expression of Bax,Bcl-2,IL-6 and TNF-α.drocyte.

关 键 词：积雪草苷 miR-342-5p 水通道蛋白3 

分 类 号：R965.2[医药卫生—药理学]