Jiang Huiyu, Wang Lin, Pan Jihong
Shandong Medicinal Biotechnology Centre, Jinan, Shandong 250022, P.R. China.
Mol Med Rep. 2015 Nov;12(5):7681-6. doi: 10.3892/mmr.2015.4376. Epub 2015 Sep 28.
The aim of the present study was to assess the effects of pro-protein convertase subtilisin/kexin type 6 (PCSK6), a proteinase implicated in the proteolytic activity of various precursor proteins and involved in the regulation of protein maturation, in fibroblast‑like synoviocytes (FLS) of a rat model of collagen‑induced arthritis (CIA). Cultured FLS from CIA models were subjected to small interfering RNA mediated PCSK6 knockdown, followed by assessment of the proliferation, invasive and migratory capacity, the secretion of inflammation factors and the cell cycle. Expression of genes associated with proliferation, invasion, migration and inflammation was detected by reverse transcription polymerase chain reaction. The results showed that PCSK6 knockdown significantly decreased the cell proliferation, invasion and migration of FLS from rats with CIA. ELISA showed an obvious decrease of tumor necrosis factor α and interleukin 1β secretion, and flow cytometric analysis revealed G0/G1 arrest of FLS following PCSK6 knockdown. Furthermore, a decrease in the mRNA levels of inflammation‑associated chemokine CXCL9, angiogenesis‑associated genes MMP‑2, MMP‑9 and NOSTRIN, hypoxia‑associated gene HIF‑1α, adhesion‑associated gene MPZL2, proliferation‑associated gene IGF‑2 and citrullination‑associated gene PADI4 was detected after PCSK6 knockdown. The results of the present study indicated that inhibition of PCSK6 may have a protective role against synovitis in rheumatoid arthritis.
本研究的目的是评估前蛋白转化酶枯草杆菌蛋白酶/kexin 6型(PCSK6)在胶原诱导性关节炎(CIA)大鼠模型的成纤维样滑膜细胞(FLS)中的作用,PCSK6是一种与多种前体蛋白的蛋白水解活性有关并参与蛋白质成熟调节的蛋白酶。对来自CIA模型的培养FLS进行小干扰RNA介导的PCSK6基因敲低,随后评估其增殖、侵袭和迁移能力、炎症因子分泌及细胞周期。通过逆转录聚合酶链反应检测与增殖、侵袭、迁移和炎症相关的基因表达。结果显示,PCSK6基因敲低显著降低了CIA大鼠FLS的细胞增殖、侵袭和迁移能力。酶联免疫吸附测定显示肿瘤坏死因子α和白细胞介素1β分泌明显减少,流式细胞术分析显示PCSK6基因敲低后FLS停滞于G0/G1期。此外,PCSK6基因敲低后,检测到炎症相关趋化因子CXCL9、血管生成相关基因MMP-2、MMP-9和NOSTRIN、缺氧相关基因HIF-1α、黏附相关基因MPZL2、增殖相关基因IGF-2和瓜氨酸化相关基因PADI4的mRNA水平降低。本研究结果表明,抑制PCSK6可能对类风湿关节炎的滑膜炎具有保护作用。