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

目的 探讨积雪草昔(Ass)对白细胞介素1B(IL-1B)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-1B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-1&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。

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

Objective To investigate the effect and mechanism of asiaticoside ( Ass ) on chondrocyte injury induced by interleukin 1p( IL-1p ) . Method After different concentrations of Ass were applied to ( IL-1 ) Ass on chondrocyte injury induced by interleukin 1p( IL-1p ) . echaniccellsapoptosis, enzyme-linked immunosorbent assay was used to detect the levels of interleukin-6 ( IL-6) and tumor necrosis factor-a ( TNF-α ) in cell culture supernatants, real-time quantitative PCR was used to detect the expression levels of miR-342-5p 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-342-5p and AQP3. After miR-342-5p 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-1p-induced chondrocyte apoptosis and the expression of IL-6 and TNF-α. Result Ass could inhibit the apoptosis rate of chondrocytes induced by IL-1p and the expression of Bax protein, IL-6 and TNF-a and miR-342-5p ( P<0. 05 ) , and promote the expression of Bcl-2 protein, AQP3 mRNA and protein ( P<0. 05 ) . miR-342-5p negatively regulated AQP3 expression in chondrocytes. After inhibiting miR- 342-5p or over expressing AQP3, the apoptosis rate of chondrocytes induced by IL-1p 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-342-5p on IL-1p-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-1rocyte apoptosis and the expression of Bax, Bcl-2, IL-6 and TNF-α. drocyte.