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24 April 2016 : Animal Research  

17β-Estradiol Protects Rat Annulus Fibrosus Cells Against Apoptosis via α1 Integrin-Mediated Adhesion to Type I Collagen: An In-vitro Study

Chun-Ming ZhaoABDE, Qian ChenACDE, Wen-Jie ZhangCF, Ai-Bing HuangBD, Wei ZhangD, Hui-Lin YangC, Zhi-Ming ZhangF

DOI: 10.12659/MSM.897906

Med Sci Monit 2016; 22:1375-1383


BACKGROUND: 17β-Estradiol (E2) has been reported to protect annulus fibrosus (AF) cells in vitro against interleukin-1β (IL-1β)-induced apoptosis in a concentration-dependent manner. However, its time-response effect remains unexplored. In addition, integrin α2/collagen II interaction has been reported to influence the apoptosis of nucleus pulposus cells in vitro. Thus, we hypothesized that integrin α1/collagen II might play a role in exerting the anti-apoptosis effect by E2. The aim of the current study was to further investigate the anti-apoptotic effect of E2 and determine the role of integrin a1/collagen II interaction.

MATERIAL AND METHODS: Rat AF cells were primary cultured and used for the following experiments. AF cells were identified by immunocytochemistry of type I collagen. Cell apoptosis was detected by fluorescence-activated cell sorter (FACS) analysis. The activity of active caspase-3 was determined by use of a caspase-3 detection kit. AF cell adhesion to type I collagen was determined by cell adhesion assay. Protein level of integrin subunit α1 was quantified by Western blot and mRNA expression was determined by real-time qPCR.

RESULTS: The immunocytochemistry of type I collagen revealed that cell purity was eligible for the following experiments with 98% of purity. FACS analysis indicated time-dependent anti-apoptosis effect of E2 at time points of 6 h, 12 h, and 24 h, which was confirmed by Caspase-3 activity. Furthermore, cell adhesion assay showed that E2 significantly increased cell binding to 95% of control, and qPCR and Western blot analysis showed that E2 effectively upregulated integrin α1. However, estrogen receptor antagonist ICI182780 prohibited the effect of E2.

CONCLUSIONS: This study shows that E2 protects against apoptosis in a time-dependent manner, and α1 integrin-mediated adhesion to collagen II is essential for estrogen-dependent anti-apoptosis in rat annulus fibrosus cells in vitro.

Keywords: Annulus Fibrosus - cytology, Blotting, Western, Cell Adhesion - drug effects, Cell Shape - drug effects, Collagen Type I - metabolism, Cytoprotection - drug effects, Estradiol - pharmacology, Integrin alpha1 - metabolism, Rats, Wistar, Real-Time Polymerase Chain Reaction



01 January 2023 : Editorial  

Editorial: Current Status of Two Adjuvanted Malaria Vaccines and the World Health Organization (WHO) Strategy to Eradicate Malaria by 2030

Dinah V. Parums
Science Editor, Medical Science Monitor, International Scientific Information, Inc., Melville, NY, USA

DOI: 10.12659/MSM.939357

Med Sci Monit 2023; 29:e939357


26 January 2023 : Editorial  

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Med Sci Monit In Press; DOI: 10.12659/MSM.939580  

19 January 2023 : Clinical Research  

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Med Sci Monit In Press; DOI: 10.12659/MSM.938892  

27 December 2022 : Clinical Research  

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Med Sci Monit 2022; 28:e938926

In Press

26 Jan 2023 : Editorial  

Editorial: The XBB.1.5 (‘Kraken’) Subvariant of Omicron SARS-CoV-2 and its Rapid Global Spread

Med Sci Monit In Press; DOI: 10.12659/MSM.939580  

20 Jan 2023 : Clinical Research  

A Study of 42 Partially Edentulous Patients with Single-Crown Restorations and Implants to Compare Bone Los...

Med Sci Monit In Press; DOI: 10.12659/MSM.939225  

20 Jan 2023 : Clinical Research  

Perfusate Neutrophil Gelatinase-Associated Lipocalin, Kidney Injury Molecular-1, Liver-Type Fatty Acid Bind...

Med Sci Monit In Press; DOI: 10.12659/MSM.938758  

19 Jan 2023 : Clinical Research  

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Med Sci Monit In Press; DOI: 10.12659/MSM.938823  

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750