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细胞周期蛋白 D1 基因沉默促进白细胞介素 1β诱导的大鼠软骨细胞凋亡。

Cyclin D1 Gene Silencing Promotes IL-1β-Induced Apoptosis in Rat Chondrocytes.

机构信息

Department of Orthopedic Surgery, Shanghai Tenth People's Hospital Affiliated to Tongji University, Shanghai, 200072, P. R. China.

Medical School, University of Texas Southwestern Medical Center, Dallas, Texas.

出版信息

J Cell Biochem. 2018 Jan;119(1):290-299. doi: 10.1002/jcb.26172. Epub 2017 Jul 4.

Abstract

This study investigated the effects of cyclin D1 gene silencing on cell proliferation and apoptosis of interleukin-1β (IL-1β)-induced osteoarthritis (OA) chondrocytes. Chondrocytes from healthy sprague-dawley rats were divided into blank, OA model (chondrocytes underwent IL-1β inducement), OA trial (chondrocytes underwent IL-1β inducement with cyclin D1-shRNA treatment), and negative control (NC; chondrocytes underwent IL-1β inducement and control-shRNA treatment) groups. Cell proliferation was assessed by CCK-8 assay, and cell cycle and apoptosis by flow cytometry. qRT-PCR and Western blotting were performed to detect cyclin D1 and apoptosis-related factors expression levels. Chondrocyte proliferation increased after 72-96 h after incubation. The OA trial group exhibited reduced cell proliferation at 48, 72, and 96 h after treatment. The OA model, OA trial, and NC groups all contained more cells arrested in G1 phase and had higher apoptosis rates than the blank group. Additionally, the OA trial group contained more cells arrested in G1 phase, with increased apoptosis rates compared to the OA model and NC groups. The OA model group had lowest expression of cyclin D1 whereas the blank group contained the highest among the four groups. qRT-PCR also showed that the OA model, OA trial, and NC groups all had increased expression levels of Bax and reduced expression levels of Bcl-2 and P53 compared to the blank group, whereby by the OA group had the most significant change. The combined evidence in our study shows that cyclin D1 gene silencing suppresses proliferation and induces apoptosis of rat chondrocytes in IL-1β-induced OA. J. Cell. Biochem. 119: 290-299, 2018. © 2017 Wiley Periodicals, Inc.

摘要

本研究探讨了细胞周期蛋白 D1 基因沉默对白细胞介素-1β(IL-1β)诱导的骨关节炎(OA)软骨细胞增殖和凋亡的影响。来自健康 Sprague-Dawley 大鼠的软骨细胞分为空白组、OA 模型组(软骨细胞经 IL-1β 诱导)、OA 试验组(软骨细胞经 IL-1β 诱导和 cyclin D1-shRNA 处理)和阴性对照组(NC;软骨细胞经 IL-1β 诱导和对照-shRNA 处理)。通过 CCK-8 法评估细胞增殖,通过流式细胞术评估细胞周期和凋亡。qRT-PCR 和 Western blot 用于检测 cyclin D1 和凋亡相关因子的表达水平。孵育 72-96 小时后,软骨细胞增殖增加。OA 试验组在处理后 48、72 和 96 小时时细胞增殖减少。OA 模型组、OA 试验组和 NC 组的 G1 期阻滞细胞比例均高于空白组,凋亡率均高于空白组。此外,OA 试验组 G1 期阻滞细胞较多,凋亡率较 OA 模型组和 NC 组高。OA 模型组 cyclin D1 表达最低,空白组最高。qRT-PCR 还显示,OA 模型组、OA 试验组和 NC 组 Bax 表达均升高,Bcl-2 和 P53 表达均降低,OA 组变化最明显。本研究的综合证据表明,cyclin D1 基因沉默抑制 IL-1β 诱导的 OA 大鼠软骨细胞增殖并诱导其凋亡。J. Cell. Biochem. 119: 290-299, 2018. © 2017 Wiley Periodicals, Inc.

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