氧化应激对盆底器官脱垂患者子宫旁韧带组织细胞细胞外基质代谢的影响

Matrix remodeling in human parametrial ligaments and fibroblasts induced by oxidative stress

目的检测盆腔器官脱垂（pelvic organ prolapse，POP）患者子宫旁韧带组织中基质代谢相关Ⅰ型胶原（collagen type Ⅰ，COL1）、弹性蛋白（elastin）和基质金属蛋白酶9（matrix metalloproteinase，MMP-9）以及氧化损伤相关8-羟基脱氧鸟苷酸（8-hydroxy-2’-deoxyguanosine urine，8-OHdG）含量的变化，检测人子宫旁韧带成纤维细胞在氧化应激作用下基质代谢相关以上各项指标的变化，为研究氧化应激在POP发生发展中的作用机制提供实验基础和理论依据。方法选择2012年10月至2014年10月在武汉大学人民医院因盆腔脏器脱垂Ⅱ-Ⅳ度（POPⅡ～POPⅣ）行子宫全切术的患者50例（POP组）和因其他良性妇科疾病行子宫切除术患者40例（对照组），采用免疫组化方法检测患者的宫旁韧带（子宫骶韧带、子宫主韧带）组织中COL1、elastin、MMP-9、8-OHdG的表达；过氧化氢（hydrogen peroxide，H2O2）处理人子宫旁韧带成纤维细胞，建立氧化应激细胞模型，Western blot检测该模型细胞中COL1、elastin和MMP-9蛋白表达。结果免疫组化结果显示，与对照组相比，POP患者宫旁韧带组织中COL1、elastin表达均降低，MMP-9和8-OHdG表达升高；Western blot与免疫组化具有相一致的结果。结论氧化应激可以引起宫旁韧带成纤维细胞细胞外基质（extracellular matrix，ECM）成分改变，可能参与POP的发生发展。

Objective To investigate the expression of collagen type Ⅰ （ COL1 ）, elastin, matrix metalloproteinase （ MMP - 9 ） and 8- hydroxy- 2＇-deoxygnanosine urine（8 -OHdG） in parametrial ligaments of patients with pelvic organ prolapsed （POP） by immunohistoehemistry. To investigate the expression of COL1, elastin, MMP -9 in human parametrial ligament fibroblast induced by oxidative stress （OS）. In order to provide laboratory basis and theoretical foundation to investigate the mechanism of OS in POP. Methods The study group consisted of 50 women with POP stage 2 or more who hospitalized in the the People＇ s Hospital of Wuhan University from October 2012 to October 2014, another 40 women who received hysterectomy for benign gynecological diseases in the same periods served as the controls. Immunohistochemistry was used to evaluate the expression of COL1 ,elastin,MMP- 9,8 -OHdG in human parametrial ligaments of the two groups. Primary cultured human parametrial ligaments were treated with exogenous hydrogen peroxide（ H2 02 ）to induce an original OS cell model, in which the expression levels of COL1, elastin and MMP- 9 were evaluated by western blot analysis. Results The results of immunohistochemistry showed a significantly lower expression of COL1 and elastin, but higher expressions of MMP - 9 and 8 - OHdG in parametrial ligaments of POP group compared to the controls. It showed the same result by western blot analysis. Conclusion OS might involve in pathophysiology of POP by contributing to extraeellular matrix（ECM） metabolic disorder.