Bisphenol A (BPA) is the most common endocrine disruptor that has toxicity to the reproductive system and male infertility. However, the underlying mechanisms of BPA’s toxicity to Sertoli cells remain poorly understood. Cellular communication network factor 5 (CCN5) is reported to regulate cell proliferation, apoptosis, and differentiation. Our study demonstrated a significant elevation of CCN5 expression in the testis of nonobstructive azoospermia patients and TM4 Sertoli cells exposed to BPA. Knockdown of CCN5 reduced apoptotic cells after BPA treatment, as determined by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling assays. Cells exposed to BPA showed increased expressions of Bax and cleaved poly(ADP-ribose) polymerase, decreased expression of Bcl-2, as well as elevated activities of caspase-3 and caspase-9 in BPA-induced TM4 cells, which were reversed by CCN5 inhibition. Loss of CCN5 declined phosphorylation of protein kinase R-like endoplasmic reticulum kinase and eukaryotic translation initiation factor 2A and decreased activating transcription factor 4 and C/EBP-homologous protein in BPA-treated cells. Furthermore, silencing CCN5 blocked BPA-induced phosphorylation of p38 and c-Jun N-terminal kinase (JNK). Administration of anisomycin, a mitogen-activated protein kinase (MAPK) activator, reversed the effects of CCN5 knockdown on BPA-induced endoplasmic reticulum (ER) stress and apoptosis. Taken together, CCN5 promotes apoptosis and ER stress in Sertoli cells exposed to BPA by activating the p38/JNK MAPK signaling pathway.
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References
1.
AdegokeEO, RahmanMS, AmjadS, et al.Environmentally relevant doses of endocrine disrupting chemicals affect male fertility by interfering with sertoli cell glucose metabolism in mice. Chemosphere, 2023; 337:139277; doi: 10.1016/j.chemosphere.2023.139277
2.
AitkenRJ. What is driving the global decline of human fertility? Need for a multidisciplinary approach to the underlying mechanisms. Front Reprod Health, 2024; 6:1364352; doi: 10.3389/frph.2024.1364352
3.
BergerT, SidhuP, TangS, et al.Are testicular cortisol and WISP2 involved in estrogen-regulated sertoli cell proliferation? Anim Reprod Sci, 2019; 207:44–51; doi: 10.1016/j.anireprosci.2019.05.014
4.
Borkham-KamphorstE, MeurerSK, WeiskirchenR. Expression and biological function of the cellular communication network factor 5 (CCN5) in primary liver cells. J Cell Commun Signal, 2023; 17(2):307–320; doi: 10.1007/s12079-023-00757-8
5.
De JongeCJ, BarrattCLR, AitkenRJ, et al.Current global status of male reproductive health. Hum Reprod Open, 2024; 2024(2):hoae017; doi: 10.1093/hropen/hoae017
6.
EhrlichS, CalafatAM, HumbletO, et al.Handling of thermal receipts as a source of exposure to bisphenol A. JAMA, 2014; 311(8):859–860; doi: 10.1001/jama.2013.283735
7.
GouldJC, LeonardLS, ManessSC, et al.Bisphenol a interacts with the estrogen receptor alpha in a distinct manner from estradiol. Mol Cell Endocrinol, 1998; 142(1–2):203–214; doi: 10.1016/s0303-7207(98)00084-7
8.
GrayMR, MalmquistJA, SullivanM, et al.CCN5 expression in mammals. II. Adult rodent tissues. J Cell Commun Signal, 2007; 1(2):145–158; doi: 10.1007/s12079-007-0013-z
9.
GrünbergJR, HammarstedtA, HedjazifarS, et al.The novel secreted adipokine WNT1-inducible signaling pathway protein 2 (WISP2) is a mesenchymal cell activator of canonical WNT. J Biol Chem, 2014; 289(10):6899–6907; doi: 10.1074/jbc.M113.511964
10.
GuoQ, JinY, ChenX, et al.NF-κB in biology and targeted therapy: New insights and translational implications. Signal Transduct Target Ther, 2024; 9(1):53; doi: 10.1038/s41392-024-01757-9
11.
HaqueI, BanerjeeS, DeA, et al.CCN5/WISP-2 promotes growth arrest of triple-negative breast cancer cells through accumulation and trafficking of p27(Kip1) via Skp2 and FOXO3a regulation. Oncogene, 2015; 34(24):3152–3163; doi: 10.1038/onc.2014.250
12.
HardingHP, ZhangY, RonD. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature, 1999; 397(6716):271–274; doi: 10.1038/16729
13.
HeH, LiX, ShenJ, et al.Bisphenol a exposure causes testicular toxicity by targeting DPY30-mediated post-translational modification of PI3K/AKT signaling in mice. Ecotoxicol Environ Saf, 2022; 243:113996; doi: 10.1016/j.ecoenv.2022.113996
14.
HerathCB, JinW, WatanabeG, et al.Adverse effects of environmental toxicants, octylphenol and bisphenol A, on male reproductive functions in pubertal rats. Endocrine, 2004; 25(2):163–172; doi: 10.1385/endo:25:2:163
HotamisligilGS, DavisRJ. Cell signaling and stress responses. Cold Spring Harb Perspect Biol, 2016; 8(10); doi: 10.1101/cshperspect.a006072
17.
IidaH, MaeharaK, DoiguchiM, et al.Bisphenol A-induced apoptosis of cultured rat sertoli cells. Reprod Toxicol, 2003; 17(4):457–464; doi: 10.1016/s0890-6238(03)00034-0
18.
InaderaH, HashimotoS, DongHY, et al.WISP-2 as a novel estrogen-responsive gene in human breast cancer cells. Biochem Biophys Res Commun, 2000; 275(1):108–114; doi: 10.1006/bbrc.2000.3276
19.
JeongD, LeeMA, LiY, et al.Matricellular protein CCN5 reverses established cardiac fibrosis. J Am Coll Cardiol, 2016; 67(13):1556–1568; doi: 10.1016/j.jacc.2016.01.030
20.
JiX, LiuT, ZhaoS, et al.WISP-2, an upregulated gene in hip cartilage from the DDH model rats, induces chondrocyte apoptosis through PPARγ in vitro. Faseb J, 2020; 34(4):4904–4917; doi: 10.1096/fj.201901915R
21.
JiangX, ChenHQ, CuiZH, et al.Low-dose and combined effects of oral exposure to bisphenol a and diethylstilbestrol on the male reproductive system in adult Sprague-Dawley rats. Environ Toxicol Pharmacol, 2016; 43:94–102; doi: 10.1016/j.etap.2016.02.014
22.
KarmakarPC, AhnJS, KimYH, et al.Paternal exposure to bisphenol-A transgenerationally impairs testis morphology, germ cell associations, and stemness properties of mouse spermatogonial stem cells. Int J Mol Sci, 2020; 21(15); doi: 10.3390/ijms21155408
23.
KarnaKK, ShinYS, ChoiBR, et al.The role of endoplasmic reticulum stress response in male reproductive physiology and pathology: A review. World J Mens Health, 2020; 38(4):484–494; doi: 10.5534/wjmh.190038
24.
KawakiH, KubotaS, SuzukiA, et al.Differential roles of CCN family proteins during osteoblast differentiation: Involvement of smad and MAPK signaling pathways. Bone, 2011; 49(5):975–989; doi: 10.1016/j.bone.2011.06.033
25.
KnapkeET, MagalhaesDP, DalvieMA, et al.Environmental and occupational pesticide exposure and human sperm parameters: A navigation guide review. Toxicology, 2022; 465:153017; doi: 10.1016/j.tox.2021.153017
26.
KoppersAJ, MitchellLA, WangP, et al.Phosphoinositide 3-kinase signalling pathway involvement in a truncated apoptotic Cascade associated with motility loss and oxidative DNA damage in human spermatozoa. Biochem J, 2011; 436(3):687–698; doi: 10.1042/bj20110114
27.
KumarR. Medical management of non-obstructive azoospermia. Clinics (Sao Paulo, Brazil), 2013; 68(Suppl 1):75–79; doi: 10.6061/clinics/2013(sup01)08
28.
LakeAC, BialikA, WalshK, et al.CCN5 is a growth arrest-specific gene that regulates smooth muscle cell proliferation and motility. Am J Pathol, 2003; 162(1):219–231; doi: 10.1016/s0002-9440(10)63813-8
29.
LakeAC, CastellotJJ.Jr., CCN5 modulates the antiproliferative effect of heparin and regulates cell motility in vascular smooth muscle cells. Cell Commun Signal, 2003; 1(1):5; doi: 10.1186/1478-811x-1-5
30.
LeeHJ, ChattopadhyayS, GongEY, et al.Antiandrogenic effects of bisphenol a and nonylphenol on the function of androgen receptor. Toxicol Sci, 2003; 75(1):40–46; doi: 10.1093/toxsci/kfg150
31.
LiL, LinW, WangZ, et al.Hormone regulation in testicular development and function. Int J Mol Sci, 2024; 25(11); doi: 10.3390/ijms25115805
32.
MeekerJD, CalafatAM, HauserR. Urinary bisphenol a concentrations in relation to serum thyroid and reproductive hormone levels in men from an infertility clinic. Environ Sci Technol, 2010a;44(4):1458–1463; doi: 10.1021/es9028292
33.
MeekerJD, EhrlichS, TothTL, et al.Semen quality and sperm DNA damage in relation to urinary bisphenol a among men from an infertility clinic. Reprod Toxicol, 2010b;30(4):532–539; doi: 10.1016/j.reprotox.2010.07.005
34.
MeiY, ThompsonMD, CohenRA, et al.Endoplasmic reticulum stress and related pathological processes. J Pharmacol Biomed Anal, 2013; 1(2):1000107.
35.
MinamiyamaY, IchikawaH, TakemuraS, et al.Generation of reactive oxygen species in sperms of rats as an earlier marker for evaluating the toxicity of endocrine-disrupting chemicals. Free Radic Res, 2010; 44(12):1398–1406; doi: 10.3109/10715762.2010.510523
36.
MolangiriA, VarmaS, MS, et al.Prenatal exposure to bisphenol S and bisphenol a differentially affects male reproductive system in the adult offspring. Food Chem Toxicol, 2022; 167:113292; doi: 10.1016/j.fct.2022.113292
37.
MrukDD, ChengCY. The mammalian Blood-Testis barrier: Its biology and regulation. Endocr Rev, 2015; 36(5):564–591; doi: 10.1210/er.2014-1101
38.
NetoFT, BachPV, NajariBB, et al.Spermatogenesis in humans and its affecting factors. Semin Cell Dev Biol, 2016; 59:10–26; doi: 10.1016/j.semcdb.2016.04.009
39.
NguyenMT, LeeMA, KimYK, et al.The matricellular protein CCN5 induces apoptosis in myofibroblasts through SMAD7-mediated inhibition of NFκB. PLoS One, 2022; 17(8):e0269735; doi: 10.1371/journal.pone.0269735
40.
NicopoullosJD, RamsayJW, AlmeidaPA, et al.Assisted reproduction in the azoospermic couple. BJOG, 2004; 111(11):1190–1203; doi: 10.1111/j.1471-0528.2004.00202.x
41.
OnoK, HanJ. The p38 signal transduction pathway: Activation and function. Cell Signal, 2000; 12(1):1–13; doi: 10.1016/s0898-6568(99)00071-6
42.
ParkB, KwonJE, ChoSM, et al.Protective effect of lespedeza cuneata ethanol extract on bisphenol A-induced testicular dysfunction in vivo and in vitro. Biomed Pharmacother, 2018; 102:76–85; doi: 10.1016/j.biopha.2018.03.045
43.
PeretzJ, VroomanL, RickeWA, et al.Bisphenol a and reproductive health: Update of experimental and human evidence, 2007-2013. Environ Health Perspect, 2014; 122(8):775–786; doi: 10.1289/ehp.1307728
44.
Pflieger-BrussS, SchuppeHC, SchillWB. The male reproductive system and its susceptibility to endocrine disrupting chemicals. Andrologia, 2004; 36(6):337–345; doi: 10.1111/j.1439-0272.2004.00641.x
45.
QiS, FuW, WangC, et al.BPA-induced apoptosis of rat sertoli cells through fas/FasL and JNKs/p38 MAPK pathways. Reprod Toxicol, 2014; 50:108–116; doi: 10.1016/j.reprotox.2014.10.013
46.
QianW, ZhuJ, MaoC, et al.Involvement of CaM-CaMKII-ERK in bisphenol A-induced sertoli cell apoptosis. Toxicology, 2014; 324:27–34; doi: 10.1016/j.tox.2014.06.001
47.
QinD, YanY, HuB, et al.Wisp2 disruption represses Cxcr4 expression and inhibits BMSCs homing to injured liver. Oncotarget, 2017; 8(58):98823–98836; doi: 10.18632/oncotarget.22006
48.
TabuchiY, TakasakiI, KondoT. Identification of genetic networks involved in the cell injury accompanying endoplasmic reticulum stress induced by bisphenol a in testicular sertoli cells. Biochem Biophys Res Commun, 2006; 345(3):1044–1050; doi: 10.1016/j.bbrc.2006.04.177
49.
TiwariD, VanageG. Mutagenic effect of bisphenol a on adult rat male germ cells and their fertility. Reprod Toxicol, 2013; 40:60–68; doi: 10.1016/j.reprotox.2013.05.013
50.
VitkuJ, HeracekJ, SosvorovaL, et al.Associations of bisphenol a and polychlorinated biphenyls with spermatogenesis and steroidogenesis in two biological fluids from men attending an infertility clinic. Environ Int, 2016; 89–90:166–173; doi: 10.1016/j.envint.2016.01.021
51.
WangC, FuW, QuanC, et al.The role of pten/akt signaling pathway involved in BPA-induced apoptosis of rat sertoli cells. Environ Toxicol, 2015; 30(7):793–802; doi: 10.1002/tox.21958
52.
WangC, QiS, LiuC, et al.Mitochondrial dysfunction and Ca(2+) overload in injured sertoli cells exposed to bisphenol A. Environ Toxicol, 2017; 32(3):823–831; doi: 10.1002/tox.22282
53.
WangL, DengL, LinN, et al.Berberine inhibits proliferation and apoptosis of vascular smooth muscle cells induced by mechanical stretch via the PDI/ERS and MAPK pathways. Life Sci, 2020; 259:118253; doi: 10.1016/j.lfs.2020.118253
54.
XiW, LeeCK, YeungWS, et al.Effect of perinatal and postnatal bisphenol a exposure to the regulatory circuits at the hypothalamus-pituitary-gonadal axis of CD-1 mice. Reprod Toxicol, 2011; 31(4):409–417; doi: 10.1016/j.reprotox.2010.12.002
55.
ZhangL, LiY, LiangC, et al.CCN5 overexpression inhibits profibrotic phenotypes via the PI3K/akt signaling pathway in lung fibroblasts isolated from patients with idiopathic pulmonary fibrosis and in an in vivo model of lung fibrosis. Int J Mol Med, 2014; 33(2):478–486; doi: 10.3892/ijmm.2013.1565
