摘要
目的探讨氧化应激对子宫韧带成纤维细胞增殖、凋亡、胶原合成以及炎症因子表达的影响及可能机制。方法将生长状况良好的第3代大鼠子宫韧带成纤维细胞分为低氧化应激组、高氧化应激组、对照组。选择0.2 mmol/L和0.8 mmol/L过氧化氢对子宫韧带成纤维细胞干预4 h,建立细胞低水平和高水平的氧化应激模型,对照组未进行任何干预。采用MTT法检测细胞增殖能力,Annexin V-FITC/PI法检测细胞凋亡情况,Western blot法检测Ⅰ型胶原、Ⅲ型胶原、炎症因子、信号通路相关蛋白的表达的蛋白水平。结果与对照组和低氧化应激组相比,高氧化应激组细胞增殖能力下降(P <0.05),凋亡率上升(P <0.05),Ⅰ型胶原、Ⅲ型胶原合成明显减少(P <0.05),白细胞介素1β、肿瘤坏死因子α和白细胞介素6的蛋白表达水平增加(P <0.05);与对照组和低氧化应激组相比,高氧化应激组p-ERK1/2、p-Akt的表达明显降低(P <0.05),而总蛋白ERK1/2、Akt的表达基本保持不变。低氧化应激组上述指标与对照组无显著差异。结论氧化应激微环境中较高浓度的过氧化氢通过抑制MAPK通路中ERK1/2表达和PI3K-Akt通路中Akt表达,引起子宫韧带成纤维细胞的增殖、胶原合成能力下降,凋亡细胞和炎症因子表达增多,参与盆腔器官脱垂的发生发展。
Objective To investigate the effects of oxidative stress on human uterine ligament fibroblast proliferation,apoptosis,collagen synthesis and expression of inflammatory factors.Methods Third generation rat uterine ligament fibroblasts were divided into low oxidative stress,high oxidative stress and control groups.Hydrogen peroxide at0.2 mmol/L and 0.8 mmol/L was applied to uterine ligament fibroblasts for 4 h to establish low and high level oxidative stress models,respectively.The methyl thiazolyl tetrazolium(MTT) assay was used to detect cell proliferation,the Annexin V-fluorescein isothiocyanate/propidium iodide(V-FITC/PI) method was applied to detect apoptosis,and Western blot was used to detect the protein levels of type Ⅰ collagen,type Ⅲ collagen,inflammatory factors and signal pathway-related proteins.Results Compared with the control and low oxidative stress groups,the proliferation ability of cells in the high oxidative stress group was decreased(P<0.05),the apoptosis rate was increased(P<0.05),the synthesis of type Ⅰ and type Ⅲ collagen was decreased(P<0.05),and the protein expression levels of interleukin(IL)-1β,tumour necrosis factor alpha(TNF-α) and IL-6 were increased(P<0.05).Compared with the control and low oxidative stress groups,phosphorylated-extracellular signal-regulated kinase 1/2(ERK1/2) and phosphorylated-protein kinase B(Akt) expression in the high oxidative stress group was decreased(P<0.05),whereas the expression of total protein ERK1/2 and Akt remained almost unchanged.There were no significant difference between the low oxidative stress and control groups.Conclusions A high hydrogen peroxide concentration in an oxidative stress microenvironment can inhibit ERK1/2 expression in the MAPK pathway and Akt expression in the phosphoinositide 3-kinase(PI3 K)/Akt pathway,Results ing in decreased uterine ligament fibroblast proliferation and collagen synthesis and increased expression of apoptotic cells and inflammatory factors,which may participate in the occurrence and development of
作者
卓然然
聂明朝
李丽红
李常虹
张芳芳
ZHUO Ranran;NIE Mingchao;LI Lihong;LI Changhong;ZHANG Fangfang(Hainan Women and Children’s Medical Center(Hainan Maternal and Child Health Hospital),Haikou 571100,China)
出处
《中国比较医学杂志》
CAS
北大核心
2021年第7期93-99,共7页
Chinese Journal of Comparative Medicine
