Wnt/β-catenin信号通路及WNK 1基因在盆腔脏器脱垂发病机制中的作用
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摘要
目的研究盆腔脏器脱垂(pelvic organ prolapse,POP)患者中Wnt经典信号通路(Wnt/β-catenin通路)、WNK 1基因的表达变化及两者相关性,初步探讨POP发生发展的可能分子机制。方法选择2017年1月至2018年5月在郑州大学第三附属医院因POP行阴式子宫切除术患者27例(POP组),及同期因妇科良性疾病行腹式或阴式子宫切除术患者21例(对照组)为研究对象。术中取子宫两侧主韧带组织,HE染色观察平滑肌束及成纤维细胞,Masson染色观察胶原纤维;WB及PCR检测通路相关蛋白(Wnt16、β-catenin、FZD 5)、WNK 1及Ⅰ型胶原蛋白(COL 1)的表达,并分析其与POP病理特征的关系,同时分析WNK 1基因与Wnt信号通路的相关性。结果①HE染色显示POP组平滑肌束萎缩断裂,成纤维细胞排列紊乱;②Masson染色可见胶原纤维明显受损、含量减少;③Western-blot及Real-TimePCR显示POP组Wnt 16、β-catenin、WNK 1、COL 1等mRNA及蛋白水平均明显下降,FZD 5 mRNA水平表达上升,蛋白水平表达无差异;WNK 1基因与Wnt通路表达趋势呈正相关;随着绝经时间延长及脱垂级别增加,β-catenin、WNK 1及COL 1表达下降。结论 POP的发生发展可能与WNK 1激酶水平及Wnt经典信号通路表达下降有关,β-catenin、WNK 1及COL 1的表达与绝经时间、脱垂程度有关。
Objective To investigate the expression of Wnt classical signaling pathway(Wnt/β-catenin pathway) and WNK 1 gene in patients with pelvic organ prolapse(POP) and the correlation between them, and to explore the possible molecular mechanism of POP development. Methods 27 patients(POP group) who underwent POP vaginal hysterectomy at The Third Affiliated Hospital of Zhengzhou University from January 2017 to May 2018, and 21 patients who underwent abdominal or vaginal hysterectomy for benign gynecological diseases in the same period(control group) were the subject of the study.The main ligament tissues on both sides of the uterus were taken during operation. Smooth muscle bundles and fibroblasts were observed by HE staining, and collagen fibers were observed by Masson staining. WB and PCR detect pathway-related proteins(Wnt16, β-catenin, FZD 5), WNK 1 and type I collagen(COL 1) expression, and analyze its relationship with pathological features of POP, and analyze the correlation between WNK 1 gene and Wnt signaling pathway.Results ① HE staining showed that the smooth muscle bundle atrophy and rupture in the POP group, and the fibroblasts were disorderly arranged;② Masson staining showed that collagen fibers were significantly damaged and the content was reduced;③ Western-blot and Real-Time PCR showed that the mRNA and protein levels of Wnt 16, β-catenin, WNK 1, COL 1, etc. in POP group decreased significantly, and the expression of FZD 5 mRNA level increased, and there was no difference in protein expression. The expression of WNK 1 gene was positively correlated with the expression of Wnt pathway. The expression of β-catenin, WNK 1 and COL 1 decreased with the prolongation of menopause and the increase of prolapsed. Conclusion The development of POP may be related to the decrease of WNK 1 kinase level and the expression of Wnt classical signaling pathway. The expression of β-catenin, WNK 1 and COL 1 is related to menopausal time and degree of prolapse.
Objective To investigate the expression of Wnt classical signaling pathway(Wnt/β-catenin pathway) and WNK 1 gene in patients with pelvic organ prolapse(POP) and the correlation between them, and to explore the possible molecular mechanism of POP development. Methods 27 patients(POP group) who underwent POP vaginal hysterectomy at The Third Affiliated Hospital of Zhengzhou University from January 2017 to May 2018, and 21 patients who underwent abdominal or vaginal hysterectomy for benign gynecological diseases in the same period(control group) were the subject of the study.The main ligament tissues on both sides of the uterus were taken during operation. Smooth muscle bundles and fibroblasts were observed by HE staining, and collagen fibers were observed by Masson staining. WB and PCR detect pathway-related proteins(Wnt16, β-catenin, FZD 5), WNK 1 and type I collagen(COL 1) expression, and analyze its relationship with pathological features of POP, and analyze the correlation between WNK 1 gene and Wnt signaling pathway.Results ① HE staining showed that the smooth muscle bundle atrophy and rupture in the POP group, and the fibroblasts were disorderly arranged;② Masson staining showed that collagen fibers were significantly damaged and the content was reduced;③ Western-blot and Real-Time PCR showed that the mRNA and protein levels of Wnt 16, β-catenin, WNK 1, COL 1, etc. in POP group decreased significantly, and the expression of FZD 5 mRNA level increased, and there was no difference in protein expression. The expression of WNK 1 gene was positively correlated with the expression of Wnt pathway. The expression of β-catenin, WNK 1 and COL 1 decreased with the prolongation of menopause and the increase of prolapsed. Conclusion The development of POP may be related to the decrease of WNK 1 kinase level and the expression of Wnt classical signaling pathway. The expression of β-catenin, WNK 1 and COL 1 is related to menopausal time and degree of prolapse.
引文
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[19] 朱军义.盆腔器官脱垂患者Wnt 16和FZD 5蛋白的异常变化 [J].中国现代医学杂志,2014,24(20):101-103.
[2] WU J-m,MATTHEWS C A,CONOVER M M,et al.Lifetime risk of stress urinary incontinence or pelvic organ prolapse surgery [J].Obstetrics & Gynecology,2014,123(6):1201-1206.
[3] WARD R M,VELEZ EDWARDS D R,EDWARDS T,et al.Genetic epidemiology of pelvic organ prolapse:a systematic review [J].American Journal of Obstetrics and Gynecology,2014,211(4):326-335.
[4] Majidinia M,Aghazadeh J,Jahanban-Esfahlani R,et al.The roles of Wnt/β-catenin pathway in tissue development and regenerative medicine [J].Journal of Cellular Physiology,2018,233(8):5598-5612.
[5] 戴毓欣,郎景和,朱兰,等.基因表达谱芯片在盆腔器官脱垂发病相关基因检测中的应用 [J].中华妇产科杂志,2010,45(5):342-347.
[6] MCCORMICK J A,ELLISON D H.The WNKs:atypical protein kinases with pleiotropic actions [J].Physiological Reviews,2011,91(1):177-219.
[7] ZHANG Y-j,ZHENG H-q,CHEN B-y,et al.WNK1 is required for proliferation induced by hypotonic challenge in rat vascular smooth muscle cells [J].Acta Pharmacologica Sinica,2017,39(1):35-47.
[8] MANDAI S,MORI T,NOMURA N,et al.WNK1 regulates skeletal muscle cell hypertrophy by modulating the nuclear localization and transcriptional activity of FOXO4 [J].Scientific Reports,2018,8(1):9101.
[9] SERYSHEVA E,MLODZIK M,JENNY A.WNKs in Wnt/β-catenin signaling [J].Cell Cycle (Georgetown,Tex.),2014,13(2):173-174.
[10] OZSVAR J,MITHIEUX S M,WANG R,et al.Elastin-based biomaterials and mesenchymal stem cells [J].Biomaterials Science,2015,3(6):800-809.
[11] RAO Shuquan,LANG Jinghe,ZHU Lan,et al.Exome sequencing identifies a novel gene,WNK1,for susceptibility to pelvic organ prolapse (POP) [J].PLOS One,2015,10(3):e0119482.
[12] CHEN H-y,LU Y,QI Y,et al.Relationship between the expressions of mitofusin-2 and procollagen in uterosacral ligament fibroblasts of postmenopausal patients with pelvic organ prolapsed [J].European Journal of Obstetrics & Gynecology,2014,174(1):141-145.
[13] 王真,史宏晖,陈娟,等.经典Wnt信号传导通路在盆腔器官脱垂发病中的作用 [J].中华医学杂志,2012,92(24):1669-1672.
[14] 杨威.机械力在盆腔器官脱垂发病机制中的研究 [D].石家庄:河北医科大学,2017.
[15] ZHANG Y,TU C,ZHANG D,et al.Genetic epidemiology of pelvic organ prolapse:a systematic review [J].American Journal of Obstetrics&Gynecology,2014,211(4):326-335.
[16] SWARUP S,PRADHANSUNDD T,VERHEYEN E M.Genome-wide identification of phospho-regulators of Wnt signaling in Drosophila [J].Development,2015,142(8):1502-1515.
[17] SERYSHEVA E,BERHANE H,GRUMOLATO L,et al.Wnk kinases are positive regulators of canonical Wnt/β-catenin signaling [J].EMBO Reports,2013,14(8):718-725.
[18] Rodan A R,Jenny A .WNK Kinases in Development and Disease [J].Current Topics in Developmental Biology,2016,123:1-47.
[19] 朱军义.盆腔器官脱垂患者Wnt 16和FZD 5蛋白的异常变化 [J].中国现代医学杂志,2014,24(20):101-103.