Role and significance of hydrogen peroxide-induced transforming growth factor beta1 expression in ligamentum flavum hypertrophy / 中国组织工程研究

ABSTRACT

BACKGROUND:The pathogenesis of ligamentum flavum hypertrophy remains poorly understood, and the expression of transforming growth factor beta1 (TGF-β1) is increased notably. Reactive oxygen species (ROS) accumulation is associated with tissue degeneration, which may accelerate the progression of ligamentum flavum hypertrophy by upregulating TGF-β1 expression. OBJECTIVE:To clarify the effect and significance of ROS H2O2-mediated up-regulation of TGF-β1 and collagen type Ⅰ in the progress of ligamentum flavum hypertrophy. METHODS:Ligamentum flavum was removed from a case of acquired lumbar disc herniation with normal ligamentum flavum during lumbar posterior decompression surgery, and then separated and cultured in vitro to the 4-6 generations, followed by exposure to H2O2 at various concentrations (0, 50, 100, 150, 200μmol/L) for 72 hours. The mRNA and protein expression levels of TGF-β1 and collagen type Ⅰ were detected by real-time PCR and western blot assay, respectively. RESULTS AND CONCLUSION:Real-time quantitative PCR showed that the mRNA expression level of TGF-β1 was significantly increased in the 150 and 200μmol/L groups (P<0.05). The mRNA expression level of collagen type Ⅰ was significantly higher in the experimental groups than that in the control group, especially in the 200μmol/L group (P<0.05). Western blot assay revealed that the protein expression levels of TGF-β1 and collagen type Ⅰ were significantly increased in a dose-dependent manner (P<0.05). These findings indicate that H2O2 may accelerate the progression of ligamentum flavum hypertrophy by up-regulating the expression levels of TGF-β1 and collagen type Ⅰ.