Gallic acid caused cultured mice TM4 Sertoli cells apoptosis and necrosis

  • Li, Wanhong (State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University) ;
  • Yue, Xiangpeng (State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University) ;
  • Li, Fadi (State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and Technology, Lanzhou University)
  • Received : 2018.04.20
  • Accepted : 2018.09.28
  • Published : 2019.05.01


Objective: The study was designed to determine the cytotoxic effect of gallic acid (GA), obtained by the hydrolysis of tannins, on mice TM4 Sertoli cells apoptosis. Methods: In the present study, non-tumorigenic mice TM4 Sertoli cells were treated with different concentrations of GA for 24 h. After treatment, cell viability was evaluated using WST-1, mitochondrial dysfunction, cells apoptosis and necrosis was detected using JC-1, Hoechst 33342 and propidium iodide staining. The expression levels of Cyclin B1, proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (BAX), and Caspase-3 were also detected by quantitative real-time polymerase chain reaction and Western-blotting. Results: The results showed that 20 to $400{\mu}M$ GA inhibited viability of TM4 Sertoli cells in a dose-dependent manner. Treatment with $400{\mu}M$ GA significantly inhibited PCNA and Cyclin B1 expression, however up-regulated BAX and Caspase-3 expression, caused mitochondrial membrane depolarization, activated Caspase-3, and induced DNA damage, thus, markedly increased the numbers of dead cells. Conclusion: Our findings showed that GA could disrupt mitochondrial function and caused TM4 cells to undergo apoptosis and necrosis.


Apoptosis;Gallic Acid;TM4 Sertoli Cell


Supported by : National Natural Science Foundation of China


  1. Fan L, Huo QG, Zhou ZM, Zhu ZG, Cai FY. Studies on the determination of Tannin content in sorghum. J Chinese Cereals Oil Assoc 2001;16:13-7.
  2. Yang JC, Du GF, Peng JZ. Comparison of cold resistance and nutritional quality of six tropical and subtropical leguminous forages during overwintering period. Partacultural Sci 2017; 34:794-801.
  3. Wang Z, Ding Y, Han XM, Guo YY. Detecting on polyphenol and tannin contents in different wine grapes form Penglai region. Sino Overseas Grapevine Wine 2015:35-7.
  4. Halvorson JJ, Kronberg SL, Hagerman AE. Effects of dietary tannins on total and extractable nutrients from manure. J Anim Sci 2017;95:3654-65.
  5. Vissers AM, Pellikaan WF, Bouwhuis A, et al. Laminaria digitata phlorotannins decrease protein degradation and methanogenesis during in vitro ruminal fermentation. J Sci Food Agric 2018;98:3644-50.
  6. Kronberg SL, Liebig MA. Condensed tannin in drinking water reduces greenhouse gas precursor urea in sheep and cattle urine. Rangeland Ecol Manag 2011;64:543-7.
  7. Ye YL, Yan JN, Cui JL, et al. Dynamic changes in amino acids, catechins, caffeine and gallic acid in green tea during withering. J Food Compost Anal 2018;66:98-108.
  8. Jiang J, Xiong YL. Natural antioxidants as food and feed additives to promote health benefits and quality of meat products: a review. Meat Sci 2016;120:107-17.
  9. You BR, Park WH. Gallic acid-induced lung cancer cell death is related to glutathione depletion as well as reactive oxygen species increase. Toxicol In Vitro 2010;24:1356-62.
  10. Abarikwu SO, Akiri OF, Durojaiye MA, Alabi AF. Combined administration of curcumin and gallic acid inhibits gallic acidinduced suppression of steroidogenesis, sperm output, antioxidant defenses and inflammatory responsive genes. J Steroid Biochem Mol Biol 2014;143:49-60.
  11. Mather JP. Establishment and characterization of two distinct mouse testicular epithelial cell lines. Biol Reprod 1980;23:243-52.
  12. Park WH, Chang MS, Yang WM, et al. Cytoprotective effect of Panax ginseng on gallic acid-induced toxicity in TM3 mouse Leydig cells. Fitoterapia 2007;78:577-9.
  13. Xie YY, Li ZZ, Lin GJ, et al. DNA interaction, cytotoxicity, apoptotic activity, cell cycle arrest, reactive oxygen species and mitochondrial membrane potential assay induced by ruthenium(II) polypyridyl complexes. Inorganica Chim Acta 2013;405:228-34.
  14. Begum P, Fugetsu B. Induction of cell death by graphene in Arabidopsis thaliana (Columbia ecotype) T87 cell suspensions. J Hazard Mater 2013;260:1032-41.
  15. Kroon PA. Bioavailability and bioefficacy of polyphenols in humans. Ι. Review of 97 bioavailablity studies. Am J Clin Nutra 2006;81(Suppl 1):230S-42S.
  16. You BR, Kim SZ, Kim SH, Park WH. Gallic acid-induced lung cancer cell death is accompanied by ROS increase and glutathione depletion. Mol Cell Biochem 2011;357:295-303.
  17. Coutinho EM. Gossypol: a contraceptive for men. Contraception 2002;65:259-63.
  18. Lamas CA, Gollucke AP, Dolder H. Grape juice concentrate (G8000((R)) ) intake mitigates testicular morphological and ultrastructural damage following cadmium intoxication. Int J Exp Pathol 2015;96:301-10.
  19. Shehab NG, Abu-Gharbieh E. Phenolic Profiling and evaluation of contraceptive effect of the ethanolic extract of salsola imbricata forssk. in male albino rats. Evid-Based Complement Alternat Med 2014;2014:Article ID 695291.
  20. Hajhosseini L, Khaki A, Merat E, Ainehchi N. Effect of rosmarinic acid on Sertoli cells apoptosis and serum antioxidant levels in rats after exposure to electromagnetic fields. Afr J Tradit Complement Altern Med 2013;10:477-80.
  21. Huang JF, Shui KJ, Li HY, Hu MY, Zhong GH. Antiproliferative effect of azadirachtin A on Spodoptera litura Sl-1 cell line through cell cycle arrest and apoptosis induced by up-regulation of p53. Pestic Biochem Physiol 2011;99:16-24.
  22. Chimento A, Sirianni R, Casaburi I, et al. 17${\beta}$-Estradiol activates GPER-and ESR1-dependent pathways inducing apoptosis in GC-2 cells, a mouse spermatocyte-derived cell line. Mol Cell Endocrinol 2012;355:49-59.
  23. Zhang G, Zeng X, Zhang R, et al. Dioscin suppresses hepatocellular carcinoma tumor growth by inducing apoptosis and regulation of TP53, BAX, BCL2 and cleaved CASP3. Phytomedicine 2016;23:1329-36.
  24. Hsu JD, Kao SH, Ou TT, et al. Gallic acid induces G2/M phase arrest of breast cancer cell MCF-7 through stabilization of p27 (Kip1) attributed to disruption of p27(Kip1)/Skp2 complex. J Agric Food Chem 2011;59:1996-2003.
  25. Ji BC, Hsu WH, Yang JS, et al. Gallic acid induces apoptosis via caspase-3 and mitochondrion-dependent pathways in vitro and suppresses lung xenograft tumor growth in vivo. J Agric Food Chem 2009;57:7596-604.
  26. Tan S, Guan X, Grun C, et al. Gallic acid induces mitotic catastrophe and inhibits centrosomal clustering in HeLa cells. Toxicol In Vitro 2015;30:506-13.
  27. Czabotar PE, Lessene G, Strasser A, Adams JM. Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy. Nat Rev Mol Cell Biol 2014;15:49-63.
  28. Owens TW, Foster FM, Valentijn A, Gilmore AP, Streuli CH. Role for X-linked Inhibitor of apoptosis protein upstream of mitochondrial permeabilization. J Biol Chem 2010;285:1081-8.
  29. Cepero E, King AM, Coffey LM, Perez RG, Boise LH. Caspase-9 and effector caspases have sequential and distinct effects on mitochondria. Oncogene 2005;24:6354-66.
  30. Park W, Chang MS, Kim H, et al. Cytotoxic effect of gallic acid on testicular cell lines with increasing $H_2O_2$ level in GC-1 spg cells. Toxicol In Vitro 2008;22:159-63.