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詹和道,姚江凌,崔红旺.连翘酯苷A对IL-1β诱导的软骨细胞损伤的影响[J].湖南中医药大学学报英文版,2023,43(8):1402-1407.[Click to copy
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| 连翘酯苷A对IL-1β诱导的软骨细胞损伤的影响 |
| 詹和道,姚江凌,崔红旺 |
| (海南医学院第一附属医院, 海南 海口 570102) |
| 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、5.0 μmol/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。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2023.08.009 |
| Received:May 29, 2023 |
| 基金项目:国家自然科学基金项目（81760620）；海南省重点研发计划项目（ZDYF2021SHFZ084）。 |
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| Effects of forsythoside A on IL-1β-induced chondrocyte injury |
| ZHAN Hedao,YAO Jiangling,CUI Hongwang |
| (The First Hospital of Hainan Medical College, Haikou, Hainan 570102, China) |
| Abstract: |
| 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 5.0 μmol/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-1β for 24 h afterwards; the si-NC and si-circSERPINE2 were separately transfected into chondrocytes which were co-treated with forsythoside A and IL-1β for 24 h afterwards. Then, the levels of IL-1β, IL-6, and TNF-α were 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. |
| Key words: forsythoside A circSERPINE2 interleukin-1β human articular chondrocytes inflammation apoptosis Bax protein Bcl-2 protein |
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