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Journal of Integrative Agriculture  2020, Vol. 19 Issue (7): 1857-1866    DOI: 10.1016/S2095-3119(19)62856-X
Special Issue: 动物医学合辑Veterninary Medicine
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
A protective role of resveratrol against the effects of immobilization stress in corpora lutea of mice in early pregnancy
Saif ULLAH*, Sheeraz MUSTAFA*, Wael ENNAB, Muhammad JAN, Muhammad SHAFIQ, Ngekure M. X. KAVITA, LÜ Zeng-peng, MAO Da-gan, SHI Fang-xiong
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China
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In the present study, we aimed to investigate a protective role for resveratrol against the effects of immobilization stress on corpora lutea (CL) of mice in early pregnancy.  A total of 45 early-pregnant mice were divided into no immobilization stress (NIS) group, immobilization stress (IS) group, and immobilization and resveratrol treatment (IS+RES) group (n=15).  Mice were immobilized in plastic tubes (50 mL) for 3 h per day during day 1 to 7 of pregnancy.  In the IS+RES group, 5 mg kg–1 d–1 of resveratrol was administered just prior to application of stress.  We analyzed apoptotic activity in CL by Western blotting analysis (WB), transmission electron microscopy (TEM), and immunohistochemistry (IHC).  Serum progesterone levels were examined with radioimmunoassay (RIA).  IHC results showed that the intensity of positive staining for Bax was increased, and for Bcl-2 was decreased in CL after IS, while resveratrol treatment reversed the positive staining for Bax and Bcl-2.  WB revealed that immobilization stress up-regulated the expression of Bax and caspase-9, and down-regulated Bcl-2 expression, while resveratrol treatment attenuated the effects of immobilization stress on the expression of Bax, Bcl-2 and caspase-9.  According to our TEM results, apoptosis as defined by chromatin condensation was found in CL after immobilization stress, while resveratrol inhibited the apoptosis.  We also demonstrated that immobilization stress decreased progesterone concentrations and ovarian expression of StAR, while resveratrol restored the concentrations of progesterone and expression of StAR back to normal.  These results indicated that immobilization stress induced luteal regression while resveratrol inhibited luteal regression, suggesting that resveratrol plays a protective role on corpora lutea of mice during early pregnancy.
Keywords:  immobilization stress        apoptosis        corpora lutea        resveratrol        pregnant mice  
Received: 11 June 2019   Accepted:
Fund: This work was supported by the National Natural Science Foundation of China (31501956 and 31572403).
Corresponding Authors:  Correspondence MAO Da-gan, E-mail:; SHI Fang-xiong, Tel: +86-25-84399112, E-mail:   
About author:  Saif ULLAH, E-mail:; * These authors contributed equally to this study.

Cite this article: 

Saif ULLAH, Sheeraz MUSTAFA, Wael ENNAB, Muhammad JAN, Muhammad SHAFIQ, Ngekure M. X. KAVITA, Lü Zeng-peng, MAO Da-gan, SHI Fang-xiong. 2020. A protective role of resveratrol against the effects of immobilization stress in corpora lutea of mice in early pregnancy. Journal of Integrative Agriculture, 19(7): 1857-1866.

Abd-Elaziz M, Moriya T, Akahira J I, Suzuki T, Sasano H. 2005. StAR and progesterone producing enzymes (3β-hydroxysteroid dehydrogenase and cholesterol side-chain cleavage cytochromes P450) in human epithelial ovarian carcinoma: Immunohistochemical and real-time PCR studies. Cancer Science, 96, 232–239.
Bai W J, Jin P J, Kuang M Q, Wei Q W, Shi F X, Davis J S, Mao D G. 2017. Temporal regulation of extracellular signal-regulated kinase 1/2 phosphorylation, heat shock protein 70 and activating transcription factor 3 during prostaglandin F-induced luteal regression in pseudopregnant rats following heat stress. Reproduction, Fertility and Development, 29, 1184–1193.
Bitgul G, Tekmen I, Keles D, Oktay G. 2013. Protective effects of resveratrol against chronic immobilization stress on testis. ISRN Urology, 2013, doi: 10.1155/2013/278720
Cabello E, Garrido P, Morán J, del Rey C G, Llaneza P, Llaneza-Suárez D, Alonso A, González C. 2015. Effects of resveratrol on ovarian response to controlled ovarian hyperstimulation in ob/ob mice. Fertility and Sterility, 103, 570–579.
Darnaudery M, Maccari S. 2008. Epigenetic programming of the stress response in male and female rats by prenatal restraint stress. Brain Research Reviews, 57, 571–585.
Demura R, Suzuki T, Nakamura S, Komatu H, Odagiri E, Demura H. 1989. Effect of immobilization stress on testosterone and inhibin in male rats. Journal of Andrology, 10, 210–213.
Druskovic M, Suput D, Milisav I. 2006. Overexpression of caspase-9 triggers its activation and apoptosis in vitro. Croatian Medical Journal, 47, 832–830.
Dudley J I, Lekli I, Mukherjee S, Das M, Bertelli A A, Das D K. 2008. Does white wine qualify for French paradox? Comparison of the cardioprotective effects of red and white wines and their constituents: resveratrol, tyrosol, and hydroxytyrosol. Journal of Agricultural and Food Chemistry, 20, 9362–9373.
EL Mohsin M M A, Fahim A T, Motawi T M, Ismail N A. 1997. Nicotine and stress: Effect on sex hormones and lipid profile in female rats. Pharmacological Research, 35, 181–187.
Elmore S. 2007. Apoptosis: A review of programmed cell death. Toxicologic Pathology, 35, 495–516.
Ennab W, Mustafa S, Wei Q, Lv Z, Kavita N M, Ullah S, Shi F. 2019. Resveratrol protects against restraint stress effects on stomach and spleen in adult male mice. Animals, 9, 736.
Ferry-Dumazet H, Garnier O, Mamani-Matsuda M, Vercauteren J, Belloc F, Billiard C, Dupouy M, Thiolat D, Kolb J P, Marit G. 2002. Resveratrol inhibits the growth and induces the apoptosis of both normal and leukemic hematopoietic cells. Carcinogenesis, 23, 1327–1333.
Hollenstein K, Janett F, Bleul U, Hassig M, Kahn W, Thun R. 2006. Influence of estradiol on adrenal activity in ovariectomized cows during acute stress. Animal Reproduction Science, 93, 292–302.
Huang R P, Adamson E D. 1993. Characterization of the DNA-binding properties of the early growth response-1 (Egr-1) transcription factor: evidence for modulation by a redox mechanism. DNA and Cell Biology, 12, 265–273.
Hung L M, Chen J K, Huang S S, Lee R S, Su M J. 2000. Cardioprotective effect of resveratrol, a natural antioxidant derived from grapes. Cardiovascular Research, 47, 549–555.
Jerome A, Thirumaran S, Kala S. 2017. Repertoire of noncoding RNAs in corpus luteum of early pregnancy in buffalo (Bubalus bubalis). Veterinary World, 10, 1129.
Jiang Y G, Tao P, Yong L, Li M C, Lin, Y H. 2008. Resveratrol reestablishes spermatogenesis after testicular injury in rats caused by 2,5-hexanedione. Chinese Medical Journal, 121, 1204–1209.
Johnson J, Bagley J, Skaznik-Wikiel M, Lee H J, Adams G B, Niikura Y, Tschudy K S, Tilly J C, Cortes M L, Forkert R. 2005. Oocyte generation in adult mammalian ovaries by putative germ cells in bone marrow and peripheral blood. Cell, 122, 303–315.
Li P, Zhou L, Zhao T, Liu X, Zhang P, Liu Y, Zheng X, Li Q. 2017. Caspase-9: Structure, mechanisms and clinical application. Oncotarget, 8, 23996.
Liu M, Yin Y, Ye  X, Zeng M, Zhao Q, Keefe D L, Liu L. 2013. Resveratrol protects against age-associated infertility in mice. Human Reproduction, 3, 707–717.
Maeyama H, Hirasawa T, Tahara Y, Obata C, Kasai H, Moriishi K, Mochizuki K, Kubota T. 2015. Maternal restraint stress during pregnancy in mice induces 11β-HSD1-associated metabolic changes in the livers of the offspring. Journal of Developmental Origins of Health and Disease, 6, 105–114.
Mairesse J, Lesage J, Breton C, Bréant B, Hahn T, Darnaudery M, Dickson S L, Seckl J, Blondeau B, Vieau D. 2007. Maternal stress alters endocrine function of the feto-placental unit in rats. American Journal of Physiology-Endocrinology and Metabolism, 292, E1526–E1533.
Marzo I, Brenner C, Zamzami N, Jürgensmeier J M, Susin S A, Vieira H L, Prévost M C, Xie Z, Matsuyama S, Reed J C. 1998. Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. Science, 281, 2027–2031.
Matikainen T, Perez G I, Jurisicova A, Pru J K, Schlezinger J J, Ryu H Y, Laine J, Sakai T, Korsmeyer S J, Casper R F. 2001. Aromatic hydrocarbon receptor-driven Bax gene expression is required for premature ovarian failure caused by biohazardous environmental chemicals. Nature Genetics, 28, 355.
Mehfooz A, Wei Q, Zheng K, Fadlalla M B, Maltasic G, Shi F. 2018. Protective roles of Rutin against restraint stress on spermatogenesis in testes of adult mice. Tissue and Cell, 50, 133–143.
Mustafa S, Wei Q, Ennab W, Lv Z, Nazar K, Siyal F A, Rodeni S, Kavita N M, Shi F. 2019. Resveratrol ameliorates testicular histopathology of mice exposed to restraint stress. Animals, 9, 743.
Oltval Z N, Milliman C L, Korsmeyer S J. 1993. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death. Cell, 74, 609–619.
Ortega I, Wong D H, Villanueva J A, Cress A B, Sokalska A, Stanley S D, Duleba A J. 2012. Effects of resveratrol on growth and function of rat ovarian granulosa cells. Fertility and Sterility, 98, 1563–1573.
Ozcan P, F?ç?c?oglu C, Y?ld?r?m OK, Ozkan F, Akkaya H, Aslan I. 2015. Protective effect of resveratrol against oxidative damage to ovarian reserve in female Sprague-Dawley rats. Reproductive Biomedicine Online, 31, 404–410.
Pacak K, Palkovits M. 2001. Stressor specificity of central neuroendocrine responses: Implications for stress-related disorders. Endocrine Reviews, 22, 502–548.
Penumathsa S V, Maulik N. 2009. Resveratrol: A promising agent in promoting cardioprotection against coronary heart disease. Canadian Journal of Physiology and Pharmacology, 87, 275–286.
Perez G I, Tilly J L. 1997. Cumulus cells are required for the increased apoptotic potential in oocytes of aged mice. Human Reproduction (Oxford, England), 12, 2781–2783.
Putcha G V, Deshmukh M, Johnson E M. 1999. BAX translocation is a critical event in neuronal apoptosis: Regulation by neuroprotectants, BCL-2, and caspases. Journal of Neuroscience, 19, 7476–7485.
Rencber S F, Ozbek S K, Eraldem?r C, Sezer Z, Kum T, Ceylan S, Guzel E. 2018. Effect of resveratrol and metformin on ovarian reserve and ultrastructure in PCOS: an experimental study. Journal of Ovarian Research, 11, 55.
Rocha C D, Soares M M, de Cássia Antonino D, Júnior J M, Mohallem R F F, Rodrigues A P R, Figueiredo J R, Beletti M E, Jacomini J O, Alves B G. 2018. Positive effect of resveratrol against preantral follicles degeneration after ovarian tissue vitrification. Theriogenology, 114, 244–251.
Stocco C, Telleria C, Gibori G. 2007. The molecular control of corpus luteum formation, function, and regression. Endocrine Reviews, 28, 117–149.
Sugino N, Nakamura Y, Okuno N, Shimamura K, Teyama T, Ishimatsu M, Kato H. 1994. Effects of restraint stress on luteal function in rats during mid-pregnancy. Reproduction, 101, 23–26.
Takiguchi S, Sugino N, Esato K, Karube-Harada A, Sakata A, Nakamura Y, Ishikawa H, Kato H. 2004. Differential regulation of apoptosis in the corpus luteum of pregnancy and newly formed corpus luteum after parturition in rats. Biology of Reproduction, 70, 313–318.
Thun R, Kaufmann C, Janett F. 1998. The influence of restraint stress on reproductive hormones in the cow. Reproduction in Domestic Animals, 33, 255–260.
Traslavina G A A, Franci C R. 2011. The CRH-R1 receptor mediates luteinizing hormone, prolactin, corticosterone and progesterone secretion induced by restraint stress in estrogen-primed rats. Brain Research, 1421, 11–19.
Tvrdá E, Kovácik A, Tusimová E, Lukác N, Lukácová J, Massányi P. 2015. Antioxidant efficiency of resveratrol on oxidative stress-induced damage in bovine spermatozoa. The Journal of Microbiology, Biotechnology and Food Sciences, 5, 64. (in Slovak)
Uguralp S, Usta U, Mizrak B. 2005. Resveratrol may reduce apoptosis of rat testicular germ cells after experimental testicular torsion. European Journal of Pediatric Surgery, 15, 333–336.
Ullah S, Zhang M, Yu H, Mustafa S, Shafiq M, Wei Q, Wang W, Jan M, Mao D. 2019. Heat exposure affected the reproductive performance of pregnant mice: Enhancement of autophagy and alteration of subcellular structure in the corpus luteum. Reproductive Biology, 19, 261–269.
Wang Y Y, Sun Y C, Sun X F, Cheng S F, Li B, Zhang X F, De Felici M, Shen W. 2017. Starvation at birth impairs germ cell cyst breakdown and increases autophagy and apoptosis in mouse oocytes. Cell Death & Disease, 8, e2613.
Wei Q, Li J, Li X, Zhang L, Shi F. 2014. Reproductive toxicity in acrylamide-treated female mice. Reproductive Toxicology, 46, 121–128.
Wu L M, Liu Y S, Tong X H, Shen N, Jin R T, Han H, Hu M H, Wang W, Zhou G X. 2012. Inhibition of follicular development induced by chronic unpredictable stress is associated with growth and differentiation factor 9 and gonadotropin in mice. Biology of Reproduction, 86, 121, 121–127.
Yazawa H, Sasagawa I, Ishigooka M, Nakada T. 1999. Effect of immobilization stress on testicular germ cell apoptosis in rats. Human Reproduction, 14, 1806–1810.
Yousuf S, Atif F, Ahmad M, Hoda N, Ishrat T, Khan B, Islam F. 2009. Resveratrol exerts its neuroprotective effect by modulating mitochondrial dysfunctions and associated cell death during cerebral ischemia. Brain Research, 1250, 242–253.
Zhang S Y, Wang J Z, Li J J, Wei D L, Sui H S, Zhang Z H, Zhou P, Tan J H. 2011. Maternal restraint stress diminishes the developmental potential of oocytes. Biology of Reproduction, 84, 672–681.
Zhou P, Lian H Y, Cui W, Wei D L, Li Q, Liu Y X, Liu X Y, Tan J H. 2012. Maternal-restraint stress increases oocyte aneuploidy by impairing metaphase I spindle assembly and reducing spindle assembly checkpoint proteins in mice. Biology of Reproduction, 86, 1–14.
Zuena A R, Mairesse J, Casolini P, Cinque C, Alema G S, Morley-Fletcher S, Chiodi V, Spagnoli L G, Gradini R, Catalani A. 2008. Prenatal restraint stress generates two distinct behavioral and neurochemical profiles in male and female rats. PLoS ONE, 3, e2170.
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