| 史纪元,姬 乐,霍斌亮,刘时璋,许兵强,傅 蔷.液泡分选蛋白4B对骨关节炎软骨细胞凋亡的调控作用[J].,2020,(14):2623-2628 |
| 液泡分选蛋白4B对骨关节炎软骨细胞凋亡的调控作用 |
| Regulation of Vacuolar Protein Sorting 4B on Chondrocyte Apoptosis in Osteoarthritis |
| 投稿时间：2019-11-12 修订日期：2019-12-08 |
| DOI：10.13241/j.cnki.pmb.2020.14.005 |
| 中文关键词: 骨关节炎 液泡分选蛋白4B 软骨 细胞凋亡 p38 MAPK信号通路 |
| 英文关键词: Osteoarthritis Vacuolar protein sorting 4B Cartilage Apoptosis p38 MAPK signaling pathway |
| 基金项目:陕西省重点研发计划项目（S2017-ZDYF-YBXM-SF-0595）；陕西省重点研发计划项目（2018ZDXM-SF-057） |
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| 中文摘要: |
| 摘要 目的：探讨液泡分选蛋白4B（VPS4B）对骨关节炎软骨细胞凋亡的调控作用。方法：通过内侧半月板部分切除加前交叉韧带切断的方法建立骨关节炎SD大鼠模型，通过RT-PCR和免疫组化检测VPS4B在大鼠关节软骨中的表达。番红O/固绿染色方法检测大鼠膝关节软骨组织形态变化。通过用10 ng/mL的IL-1β诱导人软骨肉瘤细胞SW1353 24 h来模拟骨关节炎样软骨细胞损伤，Western blot检测SW1353细胞中VPS4B、凋亡相关因子（cleaved caspase-3和cleaved PARP）和磷酸化p38的表达。转染si-RNA敲低SW1353细胞中VPS4B表达，并评估其对IL-1β诱导的SW1353细胞凋亡标记和p38 MAPK信号通路的影响。膜联蛋白V (Annexin V)和碘化丙啶(PI)染色用于检测软骨细胞凋亡。结果：VPS4B在模型组大鼠的关节软骨中明显上调（P<0.05）。IL-1β诱导24 h后，SW1353细胞中的VPS4B、cleaved caspase-3、cleaved PARP和p-p38蛋白表达水平均明显增加。而转染VPS4B-siRNA敲低VPS4B的表达后，cleaved caspase-3、cleaved PARP和p-p38蛋白表达水平均被抑制，并且抑制了IL-1β诱导细胞的凋亡率。结论：VPS4B在骨关节炎发病过程中明显上调，VPS4B的上调通过激活p38 MAPK信号通路来促进软骨细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the regulation of vacuolar protein sorting 4B(VPS4B) on chondrocyte apoptosis in osteoarthritis. Methods: The osteoarthritis SD rat model was established by partial medial meniscus resection and anterior cruciate ligament severance. The expression of VPS4B in rat articular cartilage was detected by RT-PCR and immunohistochemistry. The morphological changes of rat knee joint cartilage were detected by Safranin O/ fast green staining method. The osteoarthritis like chondrocyte injury was simulated by using 10 ng/mL IL-1 β to induce human chondrosarcoma cell SW1353 for 24 h. Western blot was used to detect the expression of VPS4B, apoptosis related factors (cleaved caspase-3 and cleaved PARP) and phosphorylated p38 in SW1353 cells. The expression of VPS4B in SW1353 cells was knocked down by transfected si-RNA, and the effect of VPS4B on IL-1 β induced apoptosis markers and p38 MAPK signaling pathway was evaluated. Annexin V and propidium iodide (PI) staining were used to detect chondrocyte apoptosis. Results: VPS4B was significantly up-regulated in the articular cartilage of the model group (P<0.05). After 24 h of IL-1β induction, the expression levels of VPS4B, cleaved caspase-3, cleaved PARP and p-p38 protein in SW1353 cells increased significantly. After transfection of VPS4B-siRNA knockdown VPS4B expression, cleaved caspase-3, cleaved PARP and p-p38 protein expression levels were inhibited, and IL-1β induced apoptosis rate was inhibited. Conclusion: VPS4B is up-regulated during the pathogenesis of osteoarthritis. Up-regulation of VPS4B promotes chondrocyte apoptosis by activating p38 MAPK signaling pathway. |
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