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中华妇幼临床医学杂志(电子版) ›› 2015, Vol. 11 ›› Issue (4) : 462 -466. doi: 10.3877/cma.j.issn.1673-5250.2015.04.006

所属专题: 文献

论著

不同程度机械力作用下人子宫旁韧带成纤维细胞8-羟基脱氧鸟苷及丙二醛水平变化的研究
赵杨, 洪莉*(), 李秉枢, 洪莎莎, 胡鸣, 杨青, 汤剑明, 李洋, 张琦钒   
  1. 430060 武汉大学人民医院妇二科
  • 收稿日期:2014-10-28 修回日期:2015-06-29 出版日期:2015-08-01
  • 通信作者: 洪莉

Research on the levels of 8- hydroxydeoxyguanosine and malondialdehyde induced by different mechanical stress in human parametrial ligament fibroblasts

Yang Zhao, Li Hong*(), Bingshu Li, Shasha Hong, Ming Hu, Qing Yang, Jianming Tang, Yang Li, Qifan Zhang   

  1. Department of Gynaecology and Obstetrics, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
  • Received:2014-10-28 Revised:2015-06-29 Published:2015-08-01
  • Corresponding author: Li Hong
引用本文:

赵杨, 洪莉, 李秉枢, 洪莎莎, 胡鸣, 杨青, 汤剑明, 李洋, 张琦钒. 不同程度机械力作用下人子宫旁韧带成纤维细胞8-羟基脱氧鸟苷及丙二醛水平变化的研究[J]. 中华妇幼临床医学杂志(电子版), 2015, 11(4): 462-466.

Yang Zhao, Li Hong, Bingshu Li, Shasha Hong, Ming Hu, Qing Yang, Jianming Tang, Yang Li, Qifan Zhang. Research on the levels of 8- hydroxydeoxyguanosine and malondialdehyde induced by different mechanical stress in human parametrial ligament fibroblasts[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2015, 11(4): 462-466.

目的

探究不同程度机械力作用下人子宫旁韧带成纤维细胞DNA氧化损伤产物8-羟基脱氧鸟苷(8-OhdG)及脂质过氧化产物丙二醛水平变化。

方法

选取2014年3月至2014年9月武汉大学人民医院妇产科收治的因非盆腔脏器脱垂(POP)的子宫良性疾病行阴式全子宫切除术患者的宫旁韧带(主韧带、骶韧带)组织标本制备的传代细胞为研究对象。根据对细胞是否加力,将其分为研究组(n=6,对黏附在爬片上的细胞进行加力)与对照组(n=3,对黏附在爬片上的细胞未加力),研究组再按照不同的加力大小,进一步分为研究A亚组(n=3,加力大小为1 333 μ)和研究B亚组(n=3,加力大小为5 333 μ)使用四点弯曲细胞力学加载系统对细胞进行力学加载,加力大小分别为0,1 333,5 333 μ,加力频率为0.1 Hz,加力时间为4 h。采用细胞免疫荧光法检测3组细胞内8-OhdG水平,硫代巴比妥酸(TBA)法检测细胞脂质过氧化产物丙二醛水平。

结果

①研究B亚组细胞内8-OhdG相对荧光强度(0.087±0.006)较研究A亚组(0.074±0.005)增强,两组分别与对照组(0.010±0.004)比较,差异均有统计学意义(t=11.286,19.633;P<0.05)。②研究A亚组细胞内丙二醛水平[(11.95± 2.07) nmoL/mgprot]较对照组[(3.18±0.50) nmoL/mgprot]无明显升高,两组比较,差异无统计学意义(t=1.509,P>0.05);而研究B亚组细胞内丙二醛水平[(65.98±11.43) nmoL/mgprot]较对照组显著升高,两组比较,差异有统计学意义(t=10.800,P<0.05);研究A组丙二醛水平与研究B亚组比较,差异有统计学意义(t=9.290,P<0.05)。

结论

一定范围内的机械力可诱导人子宫旁韧带成纤维细胞内DNA氧化损伤产物8-OhdG及脂质过氧化物丙二醛水平增加。

Objective

To explore the variation of 8-hydroxydeoxyguanosine (OhdG) and malondialdehyde (MDA) in human parametrial ligament fibroblasts induced by different mechanical stress.

Methods

Uuterosacral ligament and cardinal ligament of 10 patients with uterine benign disease but not pelvic organ prolapse (POP) who received total vaginal hysterectomy, and to derive for fibroblasts. The fibroblasts were divided into study group (n=6) and control group (n=3) according to different machenical stress, and study group were further sub-divided into two groups, study sub-group A (n=3) and study sub-group B (n=3). The fibroblasts were stretched by four-point bending system for 0, 1 333, 5 333 μ for the frequency of 0.1 Hz and 4 hours. Intracellular 8-OhdG were investigated by immunocytofluorescent and malondialdehyde were tested by the measurement of thiobarbituric acid (TBA).

Results

①The levels of 8-OhdG in study sub-group B (0.087±0.006) and study sub-group A (0.074±0.005) were significantly higher than that of control group (0.010±0.004), and those differences were significant, respectively (t=11.286, 19.633; P<0.05). ②The levels of malondialdehyde in study sub-group A was higher than that of control group [(11.95± 2.07) nmol/mgprot vs (3.18± 0.50) nmol/mgprot], but that difference was no significant (t=1.509, P>0.05), meanwhile, the level of malondialdehyde in study sub-group B was higher than that of control group [(65.98±11.43) nmol/mgprot vs (3.18± 0.50) nmol/mgprot], and that difference was significant (t=10.800, P<0.05). Furthermore, there was sgnificant difference between study sub-group A and study sub-group B (t=9.290, P<0.05).

Conclusions

Fibroblasts with increased mechanical strain loading had enhanced biomarker oxidative damage such as 8-OhdG and malondialdehyde.

表1 两组8-OhdG相对荧光强度与丙二醛水平比较(±s)
图1 盆底成纤维细胞未加力及加力后8-OhdG的细胞免疫荧光检测结果(×100)(图1A:研究A亚组;图1B:研究B亚组;图1C:对照组)注:FITC-羊抗鼠IgG抗体呈绿色,DAPI核染色呈蓝色,在倒置荧光显微镜下观察
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