GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism

Hong GU; Jia WANG; Wenwen ZHANG; Xiao YANG; Xiyue HUANG; Yingfeng ZHANG; Yanhua MAO; Changjiang LI; Qiuhong CHEN; Jinglin HUANG; Qi LIU; Ling WEI; Congcong SUN

doi:10.16016/j.2097-0927.202403072

您当前的位置：

首页 >

文章列表页 >

GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism

Basic Medicine | 更新时间：2026-01-09

- GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism
- Journal of Army Medical University Vol. 46, Issue 18, Pages: 2110-2120(2024)
- 作者机构：
  
  401331 重庆，重庆医科大学附属大学城医院妇产科
- 作者简介：
- 基金信息：
  
  the Scientific Research Project of Chongqing Education Commission(KJQN202100460);the Scientific Research Project of Chongqing Education Commission(KJQN202200462)
- DOI：10.16016/j.2097-0927.202403072
  CLC： R345;R394.6;R711.32
- Received：24 March 2024，
  
  Revised：2024-05-19，
  
  Published：30 September 2024
- 稿件说明：
移动端阅览
Hong GU, Jia WANG, Wenwen ZHANG, et al. GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism[J]. Journal of Army Medical University, 2024, 46(18): 2110-2120.
DOI：

Hong GU, Jia WANG, Wenwen ZHANG, et al. GNMT inhibits intrauterine adhesion fibrosis through TGF-β1/Smad3 signaling pathway and its mechanism[J]. Journal of Army Medical University, 2024, 46(18): 2110-2120. DOI： 10.16016/j.2097-0927.202403072.

摘要

目的

探究甘氨酸N-甲基转移酶(glycine N-methyl transferase

GNMT)对宫腔粘连(intrauterine adhesion，IUA)纤维化的影响及其相关机制。

方法

体内实验：采用36只SPF级健康雌性SD大鼠(周龄为6~8周，体质量180~220 g)。分别构建SD大鼠IUA模型及GNMT过表达大鼠IUA模型，利用RT-qPCR、免疫荧光检测正常子宫与模型组子宫GNMT表达水平；进一步利用RT-qPCR及Western blot检测各组子宫纤维化相关分子mRNA和蛋白水平以及TGF-β1/Smad3信号通路激活情况；HE染色观察各组子宫内膜腺体数量；Masson染色分析各组子宫内膜纤维化程度。体外实验：利用TGF-β1构建人子宫内膜间质细胞(transformed human endometrial stromal cells，THESCs)纤维化表型模型，使用TGF-β1处理稳定转染过表达GNMT慢病毒的THESCs，RT-qPCR及Western blot检测纤维化表型相关分子mRNA和蛋白的表达情况；Western blot检测TGF-β1/Smad3信号通路相关蛋白表达；通过TGF-β1/Smad信号通路激活剂(SRI-011381)激活TGF-β1/Smad3信号通路，Western blot检测TGF-β1/Smad3信号通路及纤维化表型关键分子蛋白表达情况。

结果

体内实验：与对照组比较，IUA大鼠GNMT mRNA和蛋白表达水平下降(

0.05)。过表达GNMT可降低IUA大鼠子宫组织纤维化相关分子Collagen Ⅰ、Collagen Ⅲ、FN mRNA及蛋白水平(

0.05)，并降低TGF-β1及其下游的Smad3蛋白磷酸化水平(

0.05)；HE和Masson染色显示过表达GNMT可增加IUA大鼠子宫内膜腺体数量并减轻纤维化程度(

0.05)。体外实验：过表达GNMT可降低THESCs纤维化表型相关分子Collagen Ⅰ、Collagen Ⅲ和FN的mRNA和蛋白水平(

0.05)，并降低TGF-β1下游的Smad3蛋白磷酸化水平(

0.05)；TGF-β1/Smad3信号通路激活后，LV-GNMT+SRI-011381组中TGF-β1/Smad3信号通路及下游的纤维化表型相关分子Collagen Ⅲ、FN的蛋白水平均明显降低。

结论

过表达GNMT通过调控TGF-β1/Smad3信号通路抑制子宫内膜纤维化，进而达到治疗IUA的作用。

Abstract

Objective

To investigate the effect of glycine N-methyl transferase (GNMT) on intrauterine adhesion (IUA) fibrosis and its related mechanism.

Methods

In vivo

experiment: A total of 36 healthy female SD rats (SPF grade

6~8 weeks old and weighing from 180~220 g) were subjected in this study. IUA model of SD rats and IUA model of GNMT overexpressed rats were established. RT-qPCR and immunofluorescence assay were applied to detect GNMT expression level in normal uterus and model group. RT-qPCR and Western blotting were used to detect the mRNA and protein levels of fibrosis-related molecules and the activation of TGF-β1/Smad3 signaling pathway in each group.

The number of endometrial glands in each group was observed by HE staining. Masson staining was used to analyze the severity of endometrial fibrosis in each group.

In vitro

experiment: transformed human endometrial stromal cells (THESCs) fibrotic phenotype model was constructed using TGF-β1

and THESCs stably transfected with GNMT overexpression lentvirus were treated with TGF-β1. RT-qPCR and Western blotting were used to detect the mRNA and protein expression of fibrosis-related molecules. The expression of TGF-β1/Smad3 signaling pathway was detected by Western blotting. TGF-β1/Smad3 signaling pathway was activated by TGF-β1/Smad signaling pathway activator (SRI-011381)

and the expression of TGF-β1/Smad3 signaling pathway and key molecular proteins of fibrosis phenotype was measured with Western blotting.

Results

In vivo

experiment

the mRNA and protein expression levels of GNMT were significantly decreased in the IUA rats than the control rats (

0.05). Overexpression of GNMT decreased the mRNA and protein levels of fibrosis related molecules

Collagen Ⅰ

Collagen Ⅲ and FN in the IUA rats (

0.05)

and decreased the phosphorylation levels of TGF-β1 and its downstream Smad3 protein (

0.05). HE and Masson staining showed that overexpression of GNMT could increase the number of endometrial glands and reduce the severity of fibrosis in the IUA rats (

0.05).

In vitro

experiments: overexpression of GNMT decreased the mRNA and protein levels of Collagen Ⅰ

Collagen Ⅲ and FN associated with fibrotic phenotype of THESCs (

0.05)

and reduced the phosphorylation level of Smad3 protein

downstream of TGF-β1 (

0.05). After activation of TGF-β1/Smad3 signaling pathway

the protein levels of TGF-β1/Smad3 signaling pathway and downstream fibrosis phenotype molecules

Collagen Ⅲ and FN

were significantly decreased in the LV

-GNMT+SRI-011381 group.

Conclusion

Overexpression of GNMT can inhibit endometrial fibrosis by regulating TGF-β1/Smad3 signaling pathway

thus achieving therapeutic effect on IUA.

关键词

Keywords

references

WU F L, LEI N J, YANG S Y, et al . Treatment strategies for intrauterine adhesion: focus on the exosomes and hydrogels[J ] . Front Bioeng Biotechnol, 2023, 11: 1264006. DOI: 10.3389/fbioe.2023.1264006.

段华, 甘露. 子宫内膜损伤与修复存在的问题和规范化管理[J]. 中国实用妇科与产科杂志, 2022, 38(9): 865-868. DOI: 10.19538/j.fk2022090101.

DUAN H, GAN L. Problems in endometrial injury and repair and the standardized management[J]. Chin J Pract Gynecol Obstet, 2022, 38(9): 865-868. DOI: 10.19538/j.fk2022090101.

桑宇超, 李淑仙, 傅松涛. 干细胞修复宫腔粘连的研究进展[J]. 现代妇产科进展, 2023, 32(6): 476-478. DOI: 10.13283/j.cnki.xdfckjz.2023.06.032.

SANG Y C, LI S X, FU S T. Research progress of stem cells in repairing intrauterine adhesions[J]. Prog Obstet Gynecol, 2023, 32(6): 476-478. DOI: 10.13283/j.cnki.xdfckjz.2023.06.032.

操媛, 王俊杰. 宫腔粘连临床治疗研究进展[J]. 实用妇科内分泌电子杂志, 2023, 10(20): 35-37.

CAO Y, WANG J J. Progress in clinical treatment of intrauterine adhesions[J]. Electron J Pract Gynecol Endocrinol, 2023, 10(20): 35-37.

俞曼殊, 史俊, 盛梅笑. DNA甲基化在纤维化疾病中的研究进展[J]. 中华中医药学刊, 2018, 36(9): 2177-2181. DOI: 10.13193/j.issn.1673-7717.2018.09.031.

YU M S, SHI J, SHENG M X. Research progress of DNA methylation in fibrotic diseases[J]. J Chin Med, 2018, 36(9): 2177-2181. DOI: 10.13193/j.issn.1673-7717.2018.09.031.

LUKA Z, PAKHOMOVA S, LOUKACHEVITCH L V, et al . Differences in folate-protein interactions result in differing inhibition of native rat liver and recombinant glycine N-methyltransferase by 5-methyltetrahydrofolate[J ] . Biochim Biophys Acta, 2012, 1824(2): 286-291. DOI: 10.1016/j.bbapap.2011.10.008.

CHANG M M, LIN C N, FANG C C, et al . Glycine N-methyltransferase inhibits aristolochic acid nephropathy by increasing CYP3A44 and decreasing NQO1 expression in female mouse hepatocytes[J ] . Sci Rep, 2018, 8(1): 6960. DOI: 10.1038/s41598-018-22298-6..

WANG Y C, LIN W L, LIN Y J, et al . A novel role of the tumor suppressor GNMT in cellular defense against DNA damage[J ] . Int J Cancer, 2014, 134(4): 799-810. DOI: 10.1002/ijc.28420.

LIU X Q, JIANG L, LEI L, et al . Carnosine alleviates diabetic nephropathy by targeting GNMT, a key enzyme mediating renal inflammation and fibrosis[J ] . Clin Sci, 2020, 134(23): 3175-3193. DOI: 10.1042/CS20201207.

FANG C C, WU C F, LIAO Y J, et al . AAV serotype 8-mediated liver specific GNMT expression delays progression of hepatocellular carcinoma and prevents carbon tetrachloride-induced liver damage[J ] . Sci Rep, 2018, 8(1): 13802. DOI: 10.1038/s41598-018-30800-3.

郭罗培, 隋龙. TGF-β在宫腔粘连纤维化中的研究进展[J]. 现代妇产科进展, 2018, 27(5): 394-396. DOI: 10.13283/j.cnki.xdfckjz.2018.05.021.

GUO L P, SUI L. Research progress of TGF-β in intrauterine adhesion fibrosis[J]. Prog Obstet Gynecol, 2018, 27(5): 394-396. DOI: 10.13283/j.cnki.xdfckjz.2018.05.021.

VANDER ARK A, CAO J C, LI X H. TGF-β receptors: in and beyond TGF-β signaling[J]. Cell Signal, 2018, 52: 112-120. DOI: 10.1016/j.cellsig.2018.09.002.

MACIAS M J, MARTIN-MALPARTIDA P, MASSAGUÉ J. Structural determinants of Smad function in TGF-β signaling[J]. Trends Biochem Sci, 2015, 40(6): 296-308. DOI: 10.1016/j.tibs.2015.03.012.

陈希琦, 张晓双, 周永坤, 等. TGF-β1/Smads信号通路在纤维化疾病中的研究进展[J]. 中国中西医结合外科杂志, 2021, 27(2): 351-354. DOI: 10.3969/j.issn.1007-6948.2021.02.037.

CHEN X Q, ZHANG X S, ZHOU Y K, et al . Research progress of TGF-β1/Smads signaling pathway in fibrotic diseases[J ] . Chin J Surg Integr Tradit West Med, 2021, 27(2): 351-354. DOI: 10.3969/j.issn.1007-6948.2021.02.037.

龚滢欣, 隋龙. 上皮间质转化在宫腔粘连中的研究进展[J]. 中国实用妇科与产科杂志, 2022, 38(5): 574-576. DOI: 10.19538/j.fk2022050122.

GONG Y X, SUI L. Research progress of epithelial-mesenchymal transition in intrauterine adhesion[J]. Chin J Pract Gynecol Obstet, 2022, 38(5): 574-576. DOI: 10.19538/j.fk2022050122.

李满超, 彭金涛, 史亚男, 等. 骨桥蛋白调控骨髓间充质干细胞抑制宫腔粘连内膜纤维化的作用研究[J]. 重庆医科大学学报, 2021, 46(8): 897-902. DOI: 10.13406/j.cnki.cyxb.002785.

LI M C, PENG J T, SHI Y N, et al . Effect of osteopontin regulating bone marrow mesenchymal stem cells on inhibiting endometrial fibrosis of intrauterine adhesions[J ] . J Chongqing Med Univ, 2021, 46(8): 897-902. DOI: 10.13406/j.cnki.cyxb.002785.

勾亚婷, 张文文, 李长江, 等. NF-κB信号通路在人羊膜间充质干细胞治疗宫腔粘连中的作用[J]. 第三军医大学学报, 2020, 42(11): 1101-1108. DOI: 10.16016/j.1000-5404.202003065.

GOU Y T, ZHANG W W, LI C J, et al . Role of NF-κB signaling pathway in treatment of intraute rine adhesions by human amniotic mesenchymal stem cells[J ] . J Third Mil Med Univ, 2020, 42(11): 1101-1108. DOI: 10.16016/j.1000-5404.202003065.

WANG K F, FANG S H, LIU Q, et al . TGF-β1/p65/MAT2A pathway regulates liver fibrogenesis via intracellular SAM[J ] . EBioMedicine, 2019, 42: 458-469. DOI: 10.1016/j.ebiom.2019.03.058.

NIETO N, CEDERBAUM A I. S-adenosylmethionine blocks collagen I production by preventing transforming growth factor-beta induction of the COL1A2 promoter[J]. J Biol Chem, 2005, 280(35): 30963-30974. DOI: 10.1074/jbc.M503569200.

SIMILE M M, BANNI S, ANGIONI E, et al . 5'-Methylthioadenosine administration prevents lipid peroxidation and fibrogenesis induced in rat liver by carbon-tetrachloride intoxication[J ] . J Hepatol, 2001, 34(3): 386-394. DOI: 10.1016/s0168-8278(00)00078-7.

杨慧楠, 吕达, 王乐, 等. TGF-β/Smad信号通路中关键基因在肺纤维化进程中的功能[J]. 环境与职业医学, 2022, 39(7): 745-751. DOI: 10.11836/JEOM21599.

YANG H N, LYU D, WANG L, et al . Functions of key genes involved in TGF-β/Smad signaling pathway in progression of pulmonary fibrosis[J ] . J Environ Occup Med, 2022, 39(7): 745-751. DOI: 10.11836/JEOM21599.

SALMA U, XUE M, ALI SHEIKH M S, et al . Role of transforming growth factor-β1 and smads signaling pathway in intrauterine adhesion[J ] . Mediators Inflamm, 2016, 2016: 4158287. DOI: 10.1155/2016/4158287.

Views

276

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Verapamil protects against hyperuricemia nephropathy through modulating TXNIP/NLRP3 inflammasome signaling pathway

Clinical effect of 2 kinds of barriers for intrauterine adhesion by hysteroscopic surgery

Effect of lncRNA-Gm4419 on proliferation and fibrosis in high glucose-induced glomerulus mesangial cells

Down-regulation of NF-κB signaling pathway promotes epithelialization of human amniotic mesenchymal stem cells

Effect of thermosensitive hydrogel containing stromal cell-derived factor-1α on intrauterine adhesion and endometrium regeneration in rabbits

Related Author

LI Li

ZHU Xin

LIU Li

ZENG Jin

YIN Zhujun

ZHAO Junning

WANG Ting

Dongmei YAO

Related Institution

School of Pharmacy， Southwest Medical University

Sichuan Academy of Chinese Medicine Sciences， Biological Assay Key Laboratory of State Administration of Traditional Chinese Medicine for Traditional Chinese Medicine Quality

Sichuan Institute for Translational Chinese Medicine

Department of Gynecology, Hubei Provincial Maternal and Children Hospital

⁰