Department of Orthopedics, Shanxi Provincial Cancer Hospital, No. 3, Zhigong New Street, Xinghualing District, Taiyuan, 030001, China.
Shanxi Health Vocational College, No. 100, Wenjin Road, Jinzhong, 030619, China.
J Orthop Sci. 2022 Jan;27(1):242-248. doi: 10.1016/j.jos.2020.12.006. Epub 2021 Feb 18.
The role of fibulin-1 or FBLN1 in chondrocyte proliferation has not been reported so far. In this study, we aimed to verify whether FBLN1 promotes chondrocyte proliferation in elderly patients with knee osteoarthritis by phosphorylating Smad2.
Chondrocytes were isolated from cartilage samples collected from elderly patients with osteoarthritis (n = 6) and young patients (n = 6). The isolated chondrocytes were divided into the following three groups: control (medium only); cells transfected with adenovirus expressing green fluorescent protein (Ad-GFP); and those transfected with adenovirus expressing green fluorescent protein and FBLN1 (Ad-GFP-FBLN1). Furthermore, chondrocytes were divided into the following three groups in the mechanistic analysis: group 1, medium only; group 2, Ad-FBLN1; and group 3, Ad-FBLN1+pSmad2 inhibitor. The cells were analyzed for the relevant indicators after culturing for 48 h.
There were more EdU-positive cells in the Ad-GFP-FBLN1 group than in the other two groups (both P < 0.05). Compared with the other two groups, the level of pSmad2 and Col2 in the Ad-GFP-FBLN1 group was significantly increased (P < 0.05). The gene expression level of each indicator was consistent with the protein expression level. There was no significant difference in the indicators between groups 1 and 3. The percentage of EdU-positive cells in group 2 was higher than that in the other two groups (P < 0.05). The expression of pSmad2 and Col2 in group 2 was higher than that in the other two groups (both P < 0.05).
FBLN1 can promote chondrocyte proliferation in the knee cartilage in elderly patients by phosphorylating Smad2.
迄今为止,纤连蛋白-1(fibulin-1,FBLN1)在软骨细胞增殖中的作用尚未见报道。本研究旨在通过磷酸化 Smad2 验证 FBLN1 是否促进老年膝骨关节炎患者的软骨细胞增殖。
从老年骨关节炎(n=6)和年轻患者(n=6)的软骨样本中分离软骨细胞。将分离的软骨细胞分为以下三组:对照组(仅培养基);转染表达绿色荧光蛋白(GFP)的腺病毒的细胞(Ad-GFP);以及转染表达 GFP 和 FBLN1 的腺病毒的细胞(Ad-GFP-FBLN1)。此外,在机制分析中,将软骨细胞分为以下三组:第 1 组,仅培养基;第 2 组,Ad-FBLN1;第 3 组,Ad-FBLN1+pSmad2 抑制剂。培养 48 h 后分析细胞的相关指标。
Ad-GFP-FBLN1 组的 EdU 阳性细胞数多于其他两组(均 P<0.05)。与其他两组相比,Ad-GFP-FBLN1 组的 pSmad2 和 Col2 水平显著增加(均 P<0.05)。各指标的基因表达水平与蛋白表达水平一致。组 1 和组 3 之间的指标无显著差异。组 2 的 EdU 阳性细胞百分比高于其他两组(P<0.05)。组 2 的 pSmad2 和 Col2 表达均高于其他两组(均 P<0.05)。
FBLN1 通过磷酸化 Smad2 促进老年膝骨关节炎患者膝关节软骨中的软骨细胞增殖。