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Original Research

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ERO1α KNOCKDOWN ATTENUATES PALMITIC ACID-MEDIATED INHIBITION OF TESTOSTERONE SECRETION BY INHIBITING ENDOPLASMIC RETICULUM STRESS IN TESTICULAR LEYDIG CELLS

  • Qi-Zhuang Lv1,2
  • Ting Qin1
  • Xin-Yun Qin1
  • Xiao-Mei Liang1
  • Ke-Yi Nong1
  • Zi-Feng Gong1
  • Li-Ying Liang1
  • Lei Yang3
  • Yu-lin Zhu1

1College of Biology & Pharmacy, Yulin Normal University, Yulin, PR China

2Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology, Yulin, PR China

3Key Laboratory of System Bio-medicine of Jiangxi Province, Jiujiang University, Jiujiang, Jiangxi, China

DOI: 10.31083/jomh.v16i3.216 Vol.16,Issue 3,July 2020 pp.75-86

Published: 16 July 2020

*Corresponding Author(s): Lei Yang E-mail: yangleigeili@163.com
*Corresponding Author(s): Yu-lin Zhu E-mail: gxzyl@163.com

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Abstract

Background and objective

Palmitic acid (PA), the most common saturated free fatty acid (FFA) in food, is related to obesity-related male infertility. The possible mechanism is PA-mediated inhibition of testosterone secretion. Endoplasmic reticulum (ER) oxidoreductin-1alpha (ERO1α), an oxidase that is localized in the ER, plays an essential role in maintaining ER homeostasis and is related to hormone secretion. However, the role and underlying mechanisms of ERO1α in PA-mediated inhibition of testosterone secretion have not been reported. 

Material and methods

Murine Leydig tumor cell line 1 (MLTC-1) cells were treated with different doses of PA. Cell viability, testosterone secretion, and ERO1α expression were measured by the Cell Counting Kit 8 (CCK-8) assay, enzyme-linked immunesorbent assay (ELISA), and Western blotting, respectively. Moreover, the expres-sion of ER stress marker proteins (glucose-regulated protein 78 [GRP78] and CCAAT/enhancer-binding protein homologous protein [CHOP]) was also measured after treatment. Subsequently, the expression ofERO1α was knocked down, and cell viability, testosterone secretion, and ER stress were measured after treatment with the PA or the ER stress agonist thapsigargin (TG, an ER stress inducer). Also, testosterone secretion was measured by ELISA when ER stress was inhibited by 4-phenylbutyric acid (4-PBA, an ER stress inhibitor).

Results

PA treatment reduced cell viability, induced ERO1α expression, and enhanced the expression of the ER stress marker GRP78 and CHOP, while ERO1α knockdown inhibits ER stress marker expression, promotes testosterone secretion, and enhances cell viability in PA-treated MLTC-1 cells. In addition, ERO1α knock-down rescued the TG-induced the decrease in testosterone secretion and cell viability.

Conclusions

These findings suggest that PA inhibits testosterone secretion via ER stress and that ERO1α knockdown ameliorates PA-induced decreases in testosterone via ER stress in testicular Leydig cells. Our results indi-cate the necessity of exploring the potential applications of ERO1α as a target gene for restoring fertility in obese men.

Keywords

endoplasmic reticulum oxidoreduclin-1 alpha; endoplasmic reticulum stress; palmitic acid; testicular Leydig cells; testosterone

Cite and Share

Qi-Zhuang Lv,Ting Qin,Xin-Yun Qin,Xiao-Mei Liang,Ke-Yi Nong,Zi-Feng Gong,Li-Ying Liang,Lei Yang,Yu-lin Zhu. ERO1α KNOCKDOWN ATTENUATES PALMITIC ACID-MEDIATED INHIBITION OF TESTOSTERONE SECRETION BY INHIBITING ENDOPLASMIC RETICULUM STRESS IN TESTICULAR LEYDIG CELLS. Journal of Men's Health. 2020. 16(3);75-86.

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