CAJ | 학술논문

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1333μ)和研究 B 亚组(n=3,加力大小为5333μ)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0,1333,5333μ,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。
목적：탐구불동정도궤계력작용하인자궁방인대성섬유세포 DNA 양화손상산물8-간기탈양조감(8-OhdG)급지질과양화산물병이철수평변화。방법선취2014년3월지2014년9월무한대학인민의원부산과수치적인비분강장기탈수(POP)적자궁량성질병행음식전자궁절제술환자적궁방인대(주인대、저인대)조직표본제비적전대세포위연구대상。근거대세포시부가력,장기분위연구조(n=6,대점부재파편상적세포진행가력)여대조조(n=3,대점부재파편상적세포미가력),연구조재안조불동적가력대소,진일보분위연구 A 아조(n=3,가력대소위1333μ)화연구 B 아조(n=3,가력대소위5333μ)사용사점만곡세포역학가재계통대세포진행역학가재,가력대소분별위0,1333,5333μ,가력빈솔위0.1 Hz,가력시간위4 h。채용세포면역형광법검측3조세포내8-OhdG 수평,류대파비타산(TBA)법검측세포지질과양화산물병이철수평。결과①연구 B 아조세포내8-OhdG 상대형광강도(0.087±0.006)교연구 A 아조(0.074±0.005)증강,량조분별여대조조(0.010±0.004)비교,차이균유통계학의의(t =11.286,19.633；P <0.05)。②연구 A 아조 세 포 내 병 이 철 수 평[(11.95±2.07)nmoL/mgprot]교대조조[(3.18±0.50)nmoL/mgprot]무명현승고,량조비교,차이무통계학의의(t =1.509,P >0.05)；이연구 B 아조세포내병이철수평[(65.98 ± 11.43)nmoL/mgprot]교대조조현저승고,량조비교,차이유통계학의의(t =10.800,P <0.05)；연구 A 조병이철수평여연구 B 아조 비 교,차이유통계학의의(t =9.290,P <0.05)。결론일정범위내적궤계력가유도인자궁방인대성섬유세포내 DNA 양화손상산물8-OhdG 급지질과양화물병이철수평증가。
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 μfor 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 ① The 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=1 1.286,1 9.633;P <0.05).②The levels of malondialdehyde in study sub-group A was higher than that of control group [(1 1.95± 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±1 1.43)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.