Department of Orthopaedics, General Hospital of Central Theater Command, Wuhan, Hubei 430070, P.R. China.
College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine, Wuhan, Hubei 430065, P.R. China.
Mol Med Rep. 2022 Sep;26(3). doi: 10.3892/mmr.2022.12805. Epub 2022 Jul 29.
The present study aimed to observe the content difference of macrophage migration inhibitory factor [MIF; novoprotein recombinant human MIF (n‑6his) (ch33)], TGFβ1 and MMP13 in patients with and without ligamentum flavum (LF) hypertrophy and investigate the roles of MIF in LF hypertrophy. The concentration of MIF, TGFβ1 and MMP13 in LF were detected by ELISA in a lumbar spinal stenosis (LSS) group and a lumbar disc herniation (LDH) group. Culture of primary LFs and identification were performed for the subsequent study. Cell treatments and cell proliferation assay by CCK‑8 was performed. Western blot and quantitative PCR analysis were used to detect the expression of TGFβ1, MMP13, type I collagen (COL‑1) and type III collagen (COL‑3) and Src which were promoted by MIF. The concentration of MIF, TGFβ1 and MMP13 were higher in the LSS group compared with the LDH group. Culture of primary LFs and identification were performed. Significant difference in LFs proliferation occurred with treatment by MIF at a concentration of 40 nM for 48 h (P<0.05). The gene and protein expression of TGFβ1, MMP13, COL‑1, COL‑3 and Src were promoted by MIF (P<0.05). Proliferation of LFs was induced by MIF and MIF‑induced proliferation of LFs was inhibited by PP1 (a Src inhibitor). MIF may promote the proliferation of LFs through the Src kinase signaling pathway and can promote extracellular matrix changes by its pro‑inflammatory effect. MIF and its mediated inflammatory reaction are driving factors of LF hypertrophy.
本研究旨在观察有和无黄韧带肥厚（LF）患者中巨噬细胞移动抑制因子 [MIF；新型蛋白重组人 MIF（n-6his）（ch33）]、TGFβ1 和 MMP13 的含量差异，并探讨 MIF 在 LF 肥厚中的作用。通过酶联免疫吸附试验（ELISA）检测腰椎管狭窄症（LSS）组和腰椎间盘突出症（LDH）组 LF 中 MIF、TGFβ1 和 MMP13 的浓度。对原代 LF 进行培养和鉴定，为后续研究做准备。用 CCK-8 进行细胞处理和细胞增殖检测。采用 Western blot 和实时定量 PCR 分析检测由 MIF 促进的 TGFβ1、MMP13、I 型胶原（COL-1）和 III 型胶原（COL-3）以及 Src 的表达。LSS 组中 MIF、TGFβ1 和 MMP13 的浓度高于 LDH 组。培养原代 LF 并进行鉴定。用 40 nM MIF 处理 48 h 后，LF 增殖有显著差异（P<0.05）。MIF 促进 TGFβ1、MMP13、COL-1、COL-3 和 Src 的基因和蛋白表达（P<0.05）。MIF 可诱导 LF 增殖，Src 激酶抑制剂 PP1 可抑制 MIF 诱导的 LF 增殖。MIF 可能通过 Src 激酶信号通路促进 LF 增殖，并通过其促炎作用促进细胞外基质的变化。MIF 及其介导的炎症反应是 LF 肥厚的驱动因素。
