连翘酯苷A对IL-1β诱导的软骨细胞损伤的影响  

Effects of forsythoside A on IL-1β-induced chondrocyte injury

作　　者：詹和道 姚江凌 崔红旺 ZHAN Hedao;YAO Jiangling;CUI Hongwang(The First Hospital of Hainan Medical College,Haikou,Hainan 570102,China)

机构地区：[1]海南医学院第一附属医院,海南海口570102

出　　处：《湖南中医药大学学报》2023年第8期1402-1407,共6页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金项目(81760620);海南省重点研发计划项目(ZDYF2021SHFZ084)。

摘　　要：目的探讨连翘酯苷A对白细胞介素-1β(interleukin-1β,IL-1β)诱导的软骨细胞损伤的影响及其可能的作用机制。方法采用50 mg/L IL-1β诱导人关节软骨细胞建立细胞损伤模型,记为模型组;另取未处理细胞为对照组。采用1.25、2.5、5.0μmol/L不同浓度的连翘酯苷A处理软骨细胞,依次记为连翘酯苷A低、中、高剂量组。ELISA法检测炎症因子[IL-1β、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)]水平;采用流式细胞术检测细胞凋亡率,qRT-PCR检测circSERPINE2表达量。pcDNA、pcDNA-circSERPINE2转染至软骨细胞后用IL-1β处理,si-NC、si-circSERPINE2分别转染至软骨细胞后用连翘酯苷A与IL-1β共同处理24 h,采用ELISA法检测IL-1β、IL-6、TNF-α的水平;流式细胞术检测细胞凋亡率;Western blot法检测Bax、Bcl-2蛋白表达量。结果连翘酯苷A作用于IL-1β诱导的软骨细胞后,炎症因子(IL-1β、IL-6、TNF-α)水平、细胞凋亡率、Bax蛋白水平降低(P<0.05),Bcl-2蛋白、circSERPINE2表达水平升高(P<0.05);转染pcDNA-circSERPINE2后炎症因子(IL-1β、IL-6、TNF-α)表达水平及细胞凋亡率、Bax蛋白水平降低(P<0.05),Bcl-2蛋白水平升高(P<0.05);转染si-circSERPINE2后,炎症因子(IL-1β、IL-6、TNF-α)水平、细胞凋亡率、Bax蛋白水平升高(P<0.05),Bcl-2蛋白水平降低(P<0.05)。结论连翘酯苷A可通过上调circSERPINE2表达而抑制细胞炎症因子表达及细胞凋亡,从而减轻IL-1β诱导的关节软骨细胞损伤。Objective To investigate the effects of forsythoside A on IL-1β-induced chondrocyte injury and its possible mechanism of action.Methods Cell injury models of Human articular chondrocytes were established using 50 mg/L interleukin-1β(IL-1β),which were denoted as model group.The untreated chondrocytes were taken as control group.Chondrocytes treated with 1.25,2.5 and 5.0μmol/L forsythoside A were recorded as low-,medium-and high-dose forsythoside A groups,respectively.The levels of inflammatory factors[IL-1β,interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)]were measured by ELISA;the apoptosis rate was determined by flow cytometry;the expression of circSERPINE2 was measured by qRT-PCR.The pcDNA and pcDNA-circSERPINE2 were separately transfected into chondrocytes which were treated with IL-1βfor 24 h afterwards;the si-NC and si-circSERPINE2 were separately transfected into chondrocytes which were co-treated with forsythoside A and IL-1βfor 24 h afterwards.Then,the levels of IL-1β,IL-6,and TNF-αwere determined by ELISA;the apoptosis rate was measured by flow cytometry;the protein expression levels of Bax and Bcl-2 were determined by Western blot.Results After forsythoside A acting on IL-1β-induced chondrocytes,the levels of inflammatory factors(IL-1β,IL-6,and TNF-α),apoptosis rate,and Bax protein level decreased(P<0.05),while the expression levels of Bcl-2 protein and circSERPINE2 increased(P<0.05).After transfection of pcDNA-circSERPINE2,the levels of inflammatory factors(IL-1β,IL-6,and TNF-α),apoptosis rate and Bax protein level decreased(P<0.05),while Bcl-2 protein level increased(P<0.05).After transfection of si-circSERPINE2,the levels of inflammatory factors(IL-1β,IL-6,and TNF-α),apoptosis rate and Bax protein level increased(P<0.05),while Bcl-2 protein level decreased(P<0.05).Conclusion Forsythoside A could reduce IL-1β-induced chondrocyte injury by up-regulating circSERPINE2 expression and thus inhibiting the expressions of cellular inflammatory factors and apoptosis.

关 键 词：连翘酯苷A circSERPINE2 白细胞介素-1Β 人关节软骨细胞 炎症 细胞凋亡 BAX蛋白 BCL-2蛋白 

分 类 号：R285.5[医药卫生—中药学]