双酚AF对人子宫内膜基质细胞衰老的影响及雷帕霉素的抑制作用

doi:10.13481/j.1671-587X.20260205

Abstract

Abstract:

Objective To investigate the effect of bisphenol AF （BPAF） on the senescence in the human endometrial stromal cells （hESCs）， and to elucidate the inhibitory effect of rapamycin （Rapa） on the BPAF-induced senescent hESCs. Methods The hESCs were cultured in vitro and treated with 0， 7.5， 15.0， 30.0， 60.0， and 120.0 μmol·L^-1 BPAF. MTT assay was used to detect the cell survival rates of the hESCs after treated with different concentrations of BPAF. The cells were divided into control group （normal medium）， BPAF group （medium containing 30.0 μmol·L^-1 BPAF）， and Rapa group （medium containing both 30.0 μmol·L^-1 BPAF and 100.0 nmol·L^-1 Rapa）. Senescence-associated β-galactosidase （SA-β-gal） staining was used to assess the percentages of SA-β-gal positive cells in various groups； flow cytometry was used to analyze the percentages of cells at different cell cycles in various groups； real-time fluorescence quantitative PCR （RT-qPCR） method was used to assess the expression levels of p16， p21 and p53 mRNA in the cells in various groups； Western blotting method was used to detect the expression levels of senescence-related proteins （p16， p21 and p53）， autophagy-related proteins p62， microtubule-associated protein 1-light chain 3 （LC3）-Ⅱ and LC3-Ⅰ in the cells in various groups. Results The MTT assay results showed that the cell survival rate was 75% when the BPAF concentration was 30.0 μmol·L^-1； therefore， this concentration was selected for subsequent senescence induction experiments. Compared with control group， the percentage of SA-β-gal positive cells in BPAF group was significantly increased（P<0.01）， the percentage of the cells at G₁ phase was significantly increased（P<0.01）， and the expression levels of p16， p21， and p53 mRNA and proteins were significantly increased （P<0.01）. Compared with BPAF group， the percentage of SA-β-gal positive cells in Rapa group was significantly decreased （P<0.01）， the percentage of the cells at G₁ phase was significantly decreased （P<0.01）， the expression levels of p16， p21， and p53 mRNA and proteins in the cells were significantly decreased （P<0.01）， and the expression level of the autophagy-related protein p62 in the cells was significantly decreased （P<0.01） while the LC3-Ⅱ/LC3-Ⅰ ratio was significantly increased （P<0.01）. Conclusion High concentrations of BPAF can inhibit the proliferation of the hESCs and induce cellular senescence. Rapa can suppress BPAF-induced cellular senescence， and its mechanism may be related with the activation of autophagy.

Key words: Endometrial stromal cell, Bisphenol AF, Cellular senescence, Rapamycin, Autophagy, Senescence-associated β-galactosidase

CLC Number:

R994.6

Ling DENG,Shouqing LI,Chunxue NIU,Xiuying LIN,Xuguang MI,Jianhua FU. Effect of bisphenol AF on senescence of human endometrial stromal cells and inhibitory role of rapamycin[J].Journal of Jilin University(Medicine Edition), 2026, 52(2): 340-347.

Figures/Tables 7

Tab.1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

References 29

[1]	GRIUKOVA A， DERYABIN P， SHATROVA A， et al. Molecular basis of senescence transmitting in the population of human endometrial stromal cells［J］. Aging， 2019， 11（21）： 9912-9931.
[2]	MARUYAMA T， YOSHIMURA Y. Molecular and cellular mechanisms for differentiation and regeneration of the uterine endometrium［J］. Endocr J， 2008， 55（5）： 795-810.
[3]	DERYABIN P I， BORODKINA A V. Stromal cell senescence contributes to impaired endometrial decidualization and defective interaction with trophoblast cells［J］. Hum Reprod， 2022， 37（7）： 1505-1524.
[4]	GORE A C， CHAPPELL V A， FENTON S E， et al. EDC-2： the endocrine society’s second scientific statement on endocrine-disrupting chemicals［J］. Endocr Rev， 2015， 36（6）： E1-E150.
[5]	WEN X， XIONG Y， GENG T， et al. BPA modulates the WDR5/TET2 complex to regulate ERβ expression in eutopic endometrium and drives the development of endometriosis［J］. Environ Pollut， 2021， 268（Pt B）： 115748.
[6]	OLSON M R， SU R W， FLAWS J A， et al. Bisphenol A impairs decidualization of human uterine stromal fibroblasts［J］. Reprod Toxicol， 2017， 73： 339-344.
[7]	HUANG M Q， LIU S， FU L， et al. Bisphenol A and its analogues bisphenol S， bisphenol F and bisphenol AF induce oxidative stress and biomacromolecular damage in human granulosa KGN cells［J］. Chemosphere， 2020， 253： 126707.
[8]	WANG B S， HAN J， ELISSEEFF J H， et al. The senescence-associated secretory phenotype and its physiological and pathological implications［J］. Nat Rev Mol Cell Biol， 2024， 25（12）： 958-978.
[9]	SELVARANI R， MOHAMMED S， RICHARDSON A. Effect of rapamycin on aging and age-related diseases-past and future［J］. Geroscience， 2021， 43（3）： 1135-1158.
[10]	LI M Y， SHEN H H， CAO X Y， et al. Targeting a mTOR/autophagy axis： a double-edged sword of rapamycin in spontaneous miscarriage［J］. Biomed Pharmacother， 2024， 177： 116976.
[11]	SHATROVA A， BUROVA E， PUGOVKINA N， et al. Monovalent ions and stress-induced senescence in human mesenchymal endometrial stem/stromal cells［J］. Sci Rep， 2022， 12（1）： 11194.
[12]	LEONE ROBERTI MAGGIORE U， CHIAPPA V， CECCARONI M， et al. Epidemiology of infertility in women with endometriosis［J］. Best Pract Res Clin Obstet Gynaecol， 2024， 92： 102454.
[13]	WANG Y D， TANG Z H， TENG X X. New advances in the treatment of thin endometrium［J］. Front Endocrinol， 2024， 15： 1269382.
[14]	WENG Z J， WANG Y G， OUCHI T， et al. Mesenchymal stem/stromal cell senescence： hallmarks， mechanisms， and combating strategies［J］. Stem Cells Transl Med， 2022， 11（4）： 356-371.
[15]	STEIN G H， DRULLINGER L F， SOULARD A， et al. Differential roles for cyclin-dependent kinase inhibitors p21 and p16 in the mechanisms of senescence and differentiation in human fibroblasts［J］. Mol Cell Biol， 1999， 19（3）： 2109-2117.
[16]	BUROVA E， BORODKINA A， SHATROVA A， et al. Sublethal oxidative stress induces the premature senescence of human mesenchymal stem cells derived from endometrium［J］. Oxid Med Cell Longev， 2013， 2013： 474931.
[17]	WU X R， LI S Q， NI Y Y， et al. Maternal BPAF exposure impaired synaptic development and caused behavior abnormality in offspring［J］. Ecotoxicol Environ Saf， 2023， 256： 114859.
[18]	CHEN D， KANNAN K， TAN H L， et al. Bisphenol analogues other than BPA： environmental occurrence， human exposure， and toxicity-a review［J］. Environ Sci Technol， 2016， 50（11）： 5438-5453.
[19]	ZHANG Y Y， LI T J， PAN C Y， et al. Intergenerational toxic effects of parental exposure to bisphenol AF on offspring and epigenetic modulations in zebrafish［J］. Sci Total Environ， 2022， 823： 153714.
[20]	LIU X X， WANG Z X， LIU F J. Chronic exposure of BPA impairs male germ cell proliferation and induces lower sperm quality in male mice［J］. Chemosphere， 2021， 262： 127880.
[21]	FAN H J， JIANG L H， LEE Y L， et al. Bisphenol compounds regulate decidualized stromal cells in modulating trophoblastic spheroid outgrowth and invasion in vitro ［J］. Biol Reprod， 2020， 102（3）： 693-704.
[22]	SIRACUSA J S， YIN L， MEASEL E， et al. Effects of bisphenol A and its analogs on reproductive health： a mini review［J］. Reprod Toxicol， 2018， 79： 96-123.
[23]	FERREIRA R， AMARAL C， CORREIA-DA-SILVA G， et al. Bisphenols A， F， S and AF trigger apoptosis and/or endoplasmic reticulum stress in human endometrial stromal cells［J］. Toxicology， 2022， 478： 153282.
[24]	MA Y， QI M， AN Y， et al. Autophagy controls mesenchymal stem cell properties and senescence during bone aging［J］. Aging Cell， 2018， 17（1）： e12709.
[25]	GARCÍA-PRAT L， MARTÍNEZ-VICENTE M， PERDIGUERO E， et al. Autophagy maintains stemness by preventing senescence［J］. Nature， 2016， 529（7584）： 37-42.
[26]	KANG C， ELLEDGE S J. How autophagy both activates and inhibits cellular senescence［J］. Autophagy， 2016， 12（5）： 898-899.
[27]	QI Z Y， YANG W Q， XUE B B， et al. ROS-mediated lysosomal membrane permeabilization and autophagy inhibition regulate bleomycin-induced cellular senescence［J］. Autophagy， 2024， 20（9）： 2000-2016.
[28]	LYU W P， LI Q H， WANG Y M， et al. Computational design of binder as the LC3-p62 protein-protein interaction［J］. Bioorg Chem， 2021， 115： 105241.
[29]	KABEYA Y， MIZUSHIMA N， UENO T， et al. LC3， a mammalian homologue of yeast Apg8p， is localized in autophagosome membranes after processing［J］. EMBO J， 2000， 19（21）： 5720-5728.

Effect of bisphenol AF on senescence of human endometrial stromal cells and inhibitory role of rapamycin

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 29

Related Articles 15

Metrics

Comments

Recommended 0

[1]	Huiyan ZHU,Min CHEN,Jinxian LI,Chunli LI. Effect of enriched environment on neurofunctional damage in rats with ischemic stroke via transcription factor EB-mediated autophagy [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 116-124.
[2]	Xiaohan YAO,Mingchen YAO,Zhiqing WANG,Heyang LI,Yan YAN,Ningjing LEI. Effect of silencing GPR139 gene on proliferation, apoptosis and autophagy of breast cancer cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 1-9.
[3]	Zhongzheng LIU,Shubo LIAN,Wenxuan FENG,Xin WEN,Hanyu LIU,Wei HE. Prometive effect of knockdown of KIF3B gene on autophagy in mouse embryonic palatal mesenchymal cells by inhibiting Shh signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1445-1451.
[4]	Haidong ZHU,Changkun LYU,Wei SHI. Effect of berberine hydrochloride on autophagy of HeLa cells infected with herpes simplex virus type 1 by regulating PI3K/AKT/mTOR signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1607-1617.
[5]	Wei AN, MAIMAITITUXUN·Tuerdi,Zhitao YAO. Regulatory effect of lutein on PI3K/AKT signaling pathway and chondrocyte autophagy in mandibular joint cartilage tissue of rabbits and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(4): 976-983.
[6]	Gao SUN,Jing HE,Qi ZHAO,Jianhong SHI,Zhiling LIAO,Yuanye TIAN,Guomin WU. Therapeutic effect of resveratrol on osteoarthritis of temporomandibular joint and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2024, 50(6): 1547-1556.
[7]	Guobin HE,Huan WANG. Effect of knockdown of RIP3 on autophagy, pyroptosis， and ferroptosis of hypoxia/reoxygenation-induced human renal tubular epithelial HK2 cells [J]. Journal of Jilin University(Medicine Edition), 2024, 50(6): 1644-1653.
[8]	Yutong WANG,Ruili RAN,Jiang BIAN,Xiaohan JIANG,Junqiu SONG,Dewei WANG,Jing YANG. Protective effect of carnosine against oxygen-glucose deprivation/reoxygenation-induced astrocyte injury through inhibition of autophagy by AMPK/mTOR signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2024, 50(5): 1297-1304.
[9]	Yuxiao SHI,Meilan LIU,Meilin ZHU,Feng WEI. Effects of 5-Aza-CdR on autophagy and apoptosis of papillary thyroid cancer cells in subcutaneous xenograft tumor tissue of nude mice and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2024, 50(5): 1330-1338.
[10]	Guoxing YU,Xin ZHANG,Hengwei DU,Bingjie CUI,Na GAO,Cuilan LIU,Jing DU. Effect of urolithin C on proliferation, apoptosis and autophagy of human acute myeloid leukemia HL-60 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2024, 50(4): 908-916.
[11]	Linru WANG,Jing ZHANG,Dongchan ZHAO,Jinjun WANG,Wenxian HU. Effect of silencing FOXO1 gene on autophagy and apoptosis of human aortic vascular smooth muscle cells [J]. Journal of Jilin University(Medicine Edition), 2024, 50(2): 431-441.
[12]	Yanhong WEI,Chenxue YANG,Guangmin YANG,Shuai SONG,Ming LI,Haijiao YANG,Haifeng WEI. Inhibitory effect of downregulating HMGB2 expression on epithelial-mesenchymal transition of liver cancer LM3 cells and its AKT/mTOR signaling pathway mechanism [J]. Journal of Jilin University(Medicine Edition), 2024, 50(1): 143-149.
[13]	Mengxue WU,Shiling CHEN,Yan LIU,Xuguang MI,Xiuying LIN,Jianhua FU,Yanqiu FANG. Effect of culture supernatant of human umbilical cord mesenchymal stem cells on survival，apoptosis and endometrium receptivity of human endometrial stromal cells after treated with mifepristone [J]. Journal of Jilin University(Medicine Edition), 2024, 50(1): 79-87.
[14]	Xiaodong WEN,Chunling WANG,Yuanjing JIANG,Xinmei ZHOU,Yi ZHANG,Yuan WU. Effect of fructus mume total flavone on injury of SH-SY5Y cells induced by MPP⁺ through regulating miR-145-3p expression and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2023, 49(6): 1415-1423.
[15]	Xing LIU,Jiali LIU,Liangui NIE,Maojun LIU,Junxiong ZHAO,Liuyang WANG,Jun YANG. Improvement effect of SO₂ on myocardial fibrosis after acute myocardial ischemic injury in rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2023, 49(5): 1125-1133.