[Animal modeling] - Relevant changes of rat uterine ligament fibroblasts induced by hydrogen peroxide in vitro

  Objective To investigate the effects of oxidative stress on the proliferation, apoptosis, collagen synthesis and the expression of inflammatory factors of uterine ligament fibroblasts.

  Methods The 3rd generation rat uterine ligament fibroblasts were divided into low oxidative stress group, high oxidative stress group and control group. 0.2 mmol/L and 0.8 mmol/L hydrogen peroxide were selected to interfere with uterine ligament fibroblasts for 4 hours to establish low level and high level oxidative stress models. The control group did not carry out any intervention. MTT method was used to detect cell proliferation, Annexin V-FITC/PI method was used to detect cell apoptosis, and Western blot method was used to detect the protein level of type I collagen, type III collagen, inflammatory factors, and signal pathway related proteins.

  Results Compared with the control group and the low oxidative stress group, the proliferation ability of cells in the high oxidative stress group decreased (P<0.05), the apoptosis rate increased (P<0.05), the synthesis of type I collagen and type III collagen decreased significantly (P<0.05), and the interleukin-1 β、 Tumor necrosis factor α The protein expression level of IL-6 and IL-6 increased (P<0.05); Compared with the control group and the low oxidative stress group, the expression of p-ERK1/2 and p-Akt in the high oxidative stress group decreased significantly (P<0.05), while the expression of total protein ERK1/2 and Akt remained unchanged. There was no significant difference between the low oxidative stress group and the control group.

  Conclusion High concentration of hydrogen peroxide in oxidative stress microenvironment can inhibit the expression of ERK1/2 in MAPK pathway and Akt in PI3K Akt pathway, reduce the proliferation and collagen synthesis of uterine ligament fibroblasts, increase the expression of apoptotic cells and inflammatory factors, and participate in the occurrence and development of pelvic organ prolapse.