Molecular Neurobiology

, Volume 54, Issue 10, pp 7620–7638 | Cite as

Ovariectomy-Induced Mitochondrial Oxidative Stress, Apoptosis, and Calcium Ion Influx Through TRPA1, TRPM2, and TRPV1 Are Prevented by 17β-Estradiol, Tamoxifen, and Raloxifene in the Hippocampus and Dorsal Root Ganglion of Rats

  • Yener Yazğan
  • Mustafa Nazıroğlu


Relative 17β-estradiol (E2) deprivation and excessive production of mitochondrial oxygen free radicals (OFRs) with a high amount of Ca2+ influx TRPA1, TRPM2, and TRPV1 activity is one of the main causes of neurodegenerative disease in postmenopausal women. In addition to the roles of tamoxifen (TMX) and raloxifene (RLX) in cancer and bone loss treatments, regulator roles in Ca2+ influx and mitochondrial oxidative stress in neurons have not been reported. The aim of this study was to evaluate whether TMX and RLX interactions with TRPA1, TRPM2, and TRPV1 in primary hippocampal (HPC) and dorsal root ganglion (DRG) neuron cultures of ovariectomized (OVX) rats. Forty female rats were divided into five groups: a control group, an OVX group, an OVX+E2 group, an OVX+TMX group, and an OVX+RLX group. The OVX+E2, OVX+TMX, and OVX+RLX groups received E2, TMX, and RLX, respectively, for 14 days after the ovariectomy. E2, ovariectomy-induced TRPA1, TRPM2, and TRPV1 current densities, as well as accumulation of cytosolic free Ca2+ in the neurons, were returned to the control levels by E2, TMX, and RLX treatments. In addition, E2, TMX, and RLX via modulation of TRPM2 and TRPV1 activity reduced ovariectomy-induced mitochondrial membrane depolarization, apoptosis, and cytosolic OFR production. TRPM2, TRPV1, PARP, and caspase-3 and caspase-9 expressions were also decreased in the neurons by the E2, TMX, and RLX treatments. In conclusion, we first reported the molecular effects of E2, TMX, and RLX on TRPA1, TRPM2, and TRPV1 channel activation in the OVX rats. In addition, we observed neuroprotective effects of E2, RLX, and TMX on oxidative and apoptotic injuries of the hippocampus and peripheral pain sensory neurons (DRGs) in the OVX rats.

Graphical Abstract

Possible molecular pathways of involvement of DEX in cerebral ischemia-induced apoptosis, oxidative stress, and calcium accumulation through TRPA1, TRPM2 and TRPV1 in the hippocampus and DRG neurons of rats. The N domain of the TRPM2 contains ADP-ribose (ADPR) pyrophosphate enzyme, which is separately activated by ADPR and oxidative stress, although the channel is reversibly inhibited by N-(p-amylcinnamoyl) anthranilic acid (ACA). The TRPV1 is also activated by mitochondrial oxidative stress and capsaicin, and it is blocked by capsazepine (CPZ). TRPA1 is also activated by oxidative stress it is inhibited by AP18. Increased cytosolic Ca2+ concentration through TRPA1, TRPM2 and TRPV1 in ovariectomized (OVX) rats may lead to neuronal toxicity, reactive oxygen species (ROS) processes, and eventual cell death. 17β-Estradiol (E2), tamoxifen (TMX), and raloxifene (RLX) reduced oxidative stress, apoptosis (including caspase-3 and caspase-9), mitochondrial membrane depolarization, and Ca2+ influx through the inhibition of TRPA1, TRPM2 and TRPV1 activation.


Hippocampus Ovariectomy Raloxifene Tamoxifen TRPM2 TRPV1 



Cytosolic free calcium ion


N-(p-Amylcinnamoyl) anthranilic acid






Cumene hydroperoxide




Dimethyl sulfoxide


Dorsal root ganglion




Ethylene glycol-bis[2-aminoethyl-ether]-N,N,N,N-tetraacetic acid




Permeability transition


Oxygen free radials


Poly(ADP-ribose) polymerase






Transient receptor potential


Transient receptor potential ankyrin 1


Transient receptor potential melastatin


Transient receptor potential vanilloid 1


Voltage-gated calcium channels


Whole cell



The abstracts of the study were presented at the 6th World Congress of Oxidative Stress, Calcium Signaling and TRP Channels, held on 24 and 27 May 2016 in Isparta, Turkey ( The authors wish to thank the researcher technicians İshak Suat Övey and Muhammet Şahin (Neuroscience Research Center, SDU, Isparta, Turkey) for helping with the Western blot and plate reader analyses.

Authorship Contributions

MN formulated the hypothesis and was responsible for writing the report. Ovariectomy of rats was performed by MN. The analyses were performed by YY.

Compliance with Ethical Standards


Unit of Scientific Research Project (BAP) of Suleyman Demirel University.

Financial Disclosure

This study was partially supported by the Unit of Scientific Research Project (BAP) of Suleyman Demirel University in Isparta, Turkey (Project Number BAP: 4135-YL2-14). There is no financial disclosure for the current study.

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Biophysics, Institute of Health ScienceSuleyman Demirel UniversityIspartaTurkey
  2. 2.Neuroscience Research CenterUniversity of Suleyman DemirelIspartaTurkey
  3. 3.Nörolojik Bilimler Uygulama ve Araştırma Merkezi MüdürüSüleyman Demirel ÜniversitesiIspartaTurkey

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