【Animal Modeling】-Hydrogen peroxide-induced changes in isolated rat uterine ligament fibroblasts

  Objective To investigate the effect and possible mechanism of oxidative stress on the proliferation, apoptosis, collagen synthesis and inflammatory factor expression of uterine ligament fibroblasts.

  Methods The third-generation rat uterine ligament fibroblasts in good growth condition were divided into low oxidative stress group, high oxidative stress group and control group. The uterine ligament fibroblasts were intervened with 0.2 mmol/L and 0.8 mmol/L hydrogen peroxide for 4 h to establish a low-level and high-level oxidative stress model, and the control group did not receive any intervention. MTT method was used to detect cell proliferation ability, Annexin V-FITC/PI method was used to detect cell apoptosis, and Western blot method was used to detect the protein levels of collagen type I, collagen type III, inflammatory factors and signaling pathway-related proteins.

  Results Compared with the control group and the low oxidative stress group, the cell proliferation ability in the high oxidative stress group decreased (P < 0. 05), the apoptosis rate increased (P < 0. 05), and the synthesis of type I collagen and type III collagen increased. significantly decreased (P < 0. 05), and the protein expression levels of interleukin-1β, tumor necrosis factor-α and interleukin-6 increased (P < 0. 05); The expressions of p-ERK1/2 and p-Akt in the oxidative stress group were significantly decreased (P < 0.05), while the expressions of total proteins ERK1/2 and Akt remained basically unchanged. There was no significant difference between the above indicators in the hypooxidative stress group and the control group.

  Conclusion Higher concentrations of hydrogen peroxide in the oxidative stress microenvironment can inhibit the expression of ERK1/2 in the MAPK pathway and the expression of Akt in the PI3K-Akt pathway. The increased expression of inflammatory factors is involved in the occurrence and development of pelvic organ prolapse.