Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter October 17, 2018

Dynamics of inflammatory reaction and oxidative stress across maternal serum, placenta and amniotic fluid in laboratory rats and the role played by genistein aglycone

Funmileyi O. Awobajo, Ayodele O. Morakinyo, Titilola A. Samuel, Oluwakemi T. Oyelowo, Abimbola O. Ogunsola, Perpetual U. Onyekwele, Mosunmola E. Okedina and Oluwadamilola O. Ogunbanwo



Genistein was reported to adversely influence fetal development although this is yet to be fully understood as a mechanism.


In this study, pregnant rats were divided into control (Cont.) and genistein force-fed (2-mg/kg and 4-mg/kg) groups. Each group was divided further into five subgroups: GD-0, GD-6, GD-13, GD-18, and GD-20 based on the terminal gestational day (GD). On the respective terminal GD, the rats were sacrificed and blood samples and amniotic fluid were carefully collected and separated and placenta homogenates were prepared. These samples were evaluated for oxidative stress and inflammatory reaction. The weights of embryonic implant and placenta tissue were also recorded. Heat shock protein (Hsp) (60 and 90), corticosterone, and oxidative stress biomarkers were determined in all the samples.


Fetal and placental weights in all genistein-exposed groups were significantly decreased. A fluctuation in the level of the Hsp was recorded with a significant decrease recorded in Hsp90 level in the placenta and amniotic fluid towards GD-20 along with a concomitant increase in the corticosterone level in the amniotic fluid in all genistein groups compared to control. Maternal serum at GD-18 and GD -20 recorded a significant increase in antioxidant level (SOD, GSH, CAT) in all genistein-exposed groups. However, these antioxidants were significantly reduced in the placenta and the amniotic fluid compared to control.


Genistein enhances the placenta function in attenuating the risk of oxidative stress in the amniotic fluid and deferentially suppressed inflammatory activities in the placenta during early gestation and towards late gestation period.


The authors appreciate the technical assistance of Mr. Folorunsho Adekunle and Mr. Dada Kayode in animal handling and sample collection. This research work was carried out at the Department of Physiology, College of Medicine of the University of Lagos, Lagos, Nigeria, with funding support through the University of Lagos CRC grant (CRC-2015/13).

  1. Author contributions: The research work was conceptualized by Dr. F.O. Awobajo. It was jointly designed by Dr. F.O. Awobajo (FOA), Dr. A.O. Morakinyo, Dr. T.A. Samuel, and Dr. O.T. Oyelowo. Investigation and acquisition of experimental data were undertaken by all authors. Manuscript was originally drafted by FOA and reviewed, edited, and approved by all authors.

  2. Research funding: University of Lagos, Nigeria (CRC No 201303 Code No 11-004-5120).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Borras C, Gambin J, Lopez-Grueso R, Pallardo FV, Vina J. Direct antioxidant and protective effect of estradiol on isolated mitochondria. BBA-Mol Basis of Dis 2010;1802:205–21.10.1016/j.bbadis.2009.09.007Search in Google Scholar

2. Jauniaux E, Watson AL, Hempstock J, Bao YP, Skepper JN, Burton GJ, et al. Onset of maternal arterial blood flow and placental oxidative stress a possible factor in human early pregnancy failure. Am J Pathol 2000;157:2111–22.10.1016/S0002-9440(10)64849-3Search in Google Scholar PubMed

3. Guerin P, El-Mouatassim S, Menezo Y. Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings. Hum Reprod Update 2001; 7:175–89.10.1093/humupd/7.2.175Search in Google Scholar PubMed

4. Harvey AJ, Kind KL. Thompson JG. REDOX regulation of early embryo development. Reproduction 2002;123:479–86.10.1530/rep.0.1230479Search in Google Scholar PubMed

5. Agarwal A, Gupta S, Sharma RK. Role of oxidative stress in female reproduction. Reproductive Biol Endocrinology 2005;3:28–37.10.1186/1477-7827-3-28Search in Google Scholar PubMed PubMed Central

6. Mor G, Cardenas I, Abrahams V, Guller S. Inflammation and pregnancy; the role of the immune system at the implantation site. Ann NY Acad Sci 2011;1221:80–7.10.1111/j.1749-6632.2010.05938.xSearch in Google Scholar PubMed PubMed Central

7. Eckert JJ, Fleming, TP. The effect of nutrition and environment on the preimplantation embryo. Obstet Gynaecol 2011;13:43–8.10.1576/toag. in Google Scholar

8. Pampfer S, Donnay I. Apoptosis at the time of embryo implantation in mouse and rat. Cell Death Differentiation 1999;6:533–45.10.1038/sj.cdd.4400516Search in Google Scholar PubMed

9. Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Kontgen F, et al. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature 1992;359:76–9.10.1038/359076a0Search in Google Scholar PubMed

10. Simon C, Gutierrez A, Vidal A, de-los-Santos MJ, Tarin JJ, Remohí J, et al. Outcome of patients with endometriosis in assisted reproduction: results from in-vitro fertilization and oocyte donation. Hum Reprod 1994;9:725–9.10.1093/oxfordjournals.humrep.a138578Search in Google Scholar PubMed

11. Awobajo FO, Nandedkar TD, Balasinor NH. Genistein alters oestrous cyclicity, oocyte fertilization and implantation process in rats. Nig Q J Hosp Med 2013;23:188–93.Search in Google Scholar

12. Awobajo FO, Onokpite BO, Ali YM, Babaleye TA, Uzor PO, Tijani KO. Genistein precipitated hypothyroidism, altered leptin and C-reactive protein synthesis in pregnant rats. Niger J Physiol Sci 2015;20;30:79–85.10.1530/endoabs.38.P144Search in Google Scholar

13. Patravale VB, Pandit NT. Design optimization of a novel method for extraction of genistein. Indian J Pharma Sci 2011;73:184–92.10.4103/0250-474X.91583Search in Google Scholar PubMed PubMed Central

14. Guide MN, Roberts CT, Kalionis B, King RG. Growth and function of the normal human placenta. Thromb Res 2004;114:397–407.10.1016/j.thromres.2004.06.038Search in Google Scholar PubMed

15. Sanin LE, López SR, Olivares ET, Terrazas MC, Silva MA, Carrillo ML. Relationship between birth weight and placenta weight. Neonatology 2001;80:113–17.10.1159/000047129Search in Google Scholar PubMed

16. Jauniaux E, Poston L, Burton GJ. Placental-related diseases of pregnancy: involvement of oxidative stress and implications in human evolution. Hum Reprod Update 2006;12:747–55.10.1093/humupd/dml016Search in Google Scholar PubMed PubMed Central

17. Negi R, Pande D, Karki K, Khanna RS, Khanna HD. Oxidative stress and preeclampsia. Adv Life Sci 2011;1:20–3.10.5923/j.als.20110101.04Search in Google Scholar

18. National Research Council (NRC). Division of earth and Life Studies. Institute for Laboratory Animal Research, Committee: guide for the care and use of laboratory animals, 8th edition. Washington, D.C.: National Academy Press, 1996: 11–124.Search in Google Scholar

19. Dinsdale EC, Ward EE. Early exposure to soy isoflavones and effects on reproductive health: a review of human and animal studies. Nutrients 2010;2:1156–87.10.3390/nu2111156Search in Google Scholar PubMed PubMed Central

20. Yagi K. Simple assay for the level of total lipid peroxides in serum or plasma. In: Armstrong D, editor. Free radical and antioxidant protocols, methods in molecular biology. Humana Press, 1998;108:101–6.10.1385/0-89603-472-0:101Search in Google Scholar

21. Tipple TE, Rogers LK. Methods for the determination of plasma or tissue glutathione levels. Methods Mol Biol 2012;889:315–24.10.1007/978-1-61779-867-2_20Search in Google Scholar PubMed PubMed Central

22. Kumar A, Dutt S, Bagler G, Ahuja PS, Kumar S. Engineering a thermo-stable superoxide dismutase functional at subzero to >50 °C, which also tolerates autoclaving. Sci Rep 2012;2:387.10.1038/srep00387Search in Google Scholar PubMed PubMed Central

23. Aebi H. Catalase in vitro. In Meth Enzymol 1984;8:121–6.10.1016/S0076-6879(84)05016-3Search in Google Scholar

24. Levy J, Faber K, Ayyash L, Hughes C. The effect of prenatal exposure to the phytoestrogen genistein on sexual differentiation in rats. Proc Soc Exp Biol Med 1995;208:60–6.10.3181/00379727-208-43832Search in Google Scholar PubMed

25. Wisniewski AB, Klein SL, Lakshmanan Y, Gearhart JP. Exposure to genistein during gestation and lactation demasculinizes the reproductive system in rats. J Urol 2003;169:1582–6.10.1097/01.ju.0000046780.23389.e0Search in Google Scholar PubMed

26. Jefferson WN, Patisaul HB, Williams CJ. Reproductive consequences of developmental phytoestrogen exposure. Reproduction 2012;143:247–60.10.1530/REP-11-0369Search in Google Scholar PubMed

27. Lumey LH. Compensatory placental growth after restricted maternal nutrition in early pregnancy. Placenta 1998; 19:105–11.10.1016/S0143-4004(98)90105-9Search in Google Scholar PubMed

28. Jansson T, Powell TL. Role of the placenta in fetal programming: underlying mechanisms and potential interventional approaches. Clin Sci (Lond) 2007;113:1–13.10.1042/CS20060339Search in Google Scholar PubMed

29. Witlin AG, Li ZY, Wimalawansa SJ, Grady JJ, Grafe MR, Yallampalli C. Placental and fetal growth and development in late rat gestation is dependent on adrenomedullin1. Biol Reprod 2002;67:1025–31.10.1095/biolreprod.101.002196Search in Google Scholar

30. Wisdom SJ, Wilson R, McKillop JH, Walker JJ. Antioxidant systems in normal pregnancy and in pregnancy-induced hypertension. Am J Obstet Gynecol 1991;165:1701–4.10.1016/0002-9378(91)90018-MSearch in Google Scholar PubMed

31. Leal CA, Schetinger MR, Leal DB, Morsch VM, da Silva AS, Rezer JF, et al. Oxidative stress and antioxidant defences in pregnant women. Redox Rep 2011;16:230–6.10.1179/1351000211Y.0000000013Search in Google Scholar PubMed PubMed Central

32. Kurlak LO, Green A, Loughna P, Pipkin FB. Oxidative stress markers in hypertensive states of pregnancy: preterm and term disease. Front Physiol 2014;5:1–8.10.3389/fphys.2014.00310Search in Google Scholar PubMed PubMed Central

33. Burton GJ, Jauniaux E. Placental oxidative stress: from miscarriage to preeclampsia. J Soc Gynecol Investig 2004;11:342–52.10.1016/j.jsgi.2004.03.003Search in Google Scholar PubMed

34. Many A, Hubel CA, Fisher SJ, Roberts JM, Zhou Y. Invasive cytotrophoblasts manifest evidence of oxidative stress in preeclampsia. Am J Pathol 2000;156:321–31.10.1016/S0002-9440(10)64733-5Search in Google Scholar PubMed

35. Raijmakers MT, Peters WH, Steegers EA, Poston L. Amino thiols, detoxification and oxidative stress in preeclampsia and other disorders of pregnancy. Curr Pharm Des 2005;11:711–34.10.2174/1381612053381837Search in Google Scholar PubMed

36. Lee DY, Kim E, Choi MH. Technical and clinical aspects of cortisol as a biochemical marker of chronic stress. BMB Rep 2015;48:209–16.10.5483/BMBRep.2015.48.4.275Search in Google Scholar PubMed

37. Burton GJ, Yung HW, Cindrova-Davies T, Charnock-Jones DS. Placental endoplasmic reticulum stress and oxidative stress in the pathophysiology of unexplained intrauterine growth restriction and early onset preeclampsia. Placenta 2009;30(A):S43–8.10.1016/j.placenta.2008.11.003Search in Google Scholar PubMed PubMed Central

38. LeWinn KJ, Stroud LR, Molnar BE, Ware JH, Koenen KC, Buka SL. Elevated maternal cortisol levels during pregnancy are associated with reduced childhood IQ. Int J Epidemiol 2009;38:1700–10.10.1093/ije/dyp200Search in Google Scholar PubMed PubMed Central

39. Davis EP, Sandman CA. The timing of prenatal exposure to maternal cortisol and psychosocial stress is associated with human infant cognitive development. Child Dev 2010;81:131–48.10.1111/j.1467-8624.2009.01385.xSearch in Google Scholar PubMed PubMed Central

40. Benediktsson R, Calder AA, Edwards CR, Seckl JR. Placental 11 betahydroxysteroid dehydrogenase: a key regulator of fetal glucocorticoid exposure. Clin Endocrinol (Oxf) 1997;46:161–6.10.1046/j.1365-2265.1997.1230939.xSearch in Google Scholar PubMed

41. Levitt NS, Lindsay RS, Holmes MC, Seckl JR. Dexamethasone in the last week of pregnancy attenuates hippocampal glucocorticoid receptor gene expression and elevates blood pressure in the adult offspring in the rat. Neuroendocrinology 1996;64:412–8.10.1159/000127146Search in Google Scholar PubMed

42. Nyirenda MJ, Seck JR. Intrauterine events and the programming of dulthood disease: the role of fetal glucocorticoid exposure. Int J Mol Med 1998;2:607–14.10.3892/ijmm.2.5.607Search in Google Scholar PubMed

43. Bloom SL, Sheffield JS, McIntire DD, Leveno KJ. Antenatal dexamethasone and decreased birth weight. Obstet Gynecol 2001;97:485–90.10.1097/00006250-200104000-00001Search in Google Scholar PubMed

44. Langdown ML, Sugden MC. Enhanced placental GLUT1 and GLUT3 expression in dexamethasone-induced fetal growth retardation. Mol Cell Endocrinol 2001;185:109–17.10.1016/S0303-7207(01)00629-3Search in Google Scholar PubMed

45. Barker DJ. Fetal nutrition and cardiovascular disease in later life. Br Med Bull 1997;53:96–108.10.1093/oxfordjournals.bmb.a011609Search in Google Scholar PubMed

46. Sugden MC, Langdown ML, Munns MJ, Holness MJ. Maternal glucocorticoid treatment modulates placental leptin and leptin receptor expression and materno-fetal leptin physiology during late pregnancy, and elicits hypertension associated with hyperleptinaemia in the early-growth-retarded adult offspring. Eur J Endocrinol 2001;145:529–39.10.1530/eje.0.1450529Search in Google Scholar PubMed

47. Pijnenborg R, Dixon G, Robertson WB, Brosens I. Trophoblastic invasion of human decidua from 8 to 18 weeks of pregnancy. Placenta 1980;1:3–19.10.1016/S0143-4004(80)80012-9Search in Google Scholar PubMed

48. Belhia F, Gremlich S, Muller-Brochut AC, Damnon F, Hhlfeld P, Witkin SS, et al. Anti-60-kDa heat shock protein antibodies in fetal serum: a biomarker for unexplained small for gestational age foetuses. Gynecol Obstet Invest 2010;70:229–305.10.1159/000314021Search in Google Scholar PubMed

49. Zugel U, Kaufmann SH. Role of heat shock proteins in protection from and pathogenesis of infectious diseases. Clin Microbiol Rev 1999;12:319–91.10.1128/CMR.12.1.19Search in Google Scholar PubMed PubMed Central

50. Ziegert M, Witkin SS, Sziller I, Alexander H, Brylla E, Hartig W. Heat shock proteins and heat shock protein antibody complexes in placenta tissue. Infectious Dis Obstetrics Gynecol 1999;7:180–5.10.1155/S1064744999000307Search in Google Scholar

Received: 2018-04-15
Accepted: 2018-07-11
Published Online: 2018-10-17
Published in Print: 2018-12-19

©2019 Walter de Gruyter GmbH, Berlin/Boston

Scroll Up Arrow