ISSN 2415-3060 (print), ISSN 2522-4972 (online)
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УЖМБС 2019, 4(1): 90–95
https://doi.org/10.26693/jmbs04.01.090
Experimental Medicine and Morphology

Immunohistochemical Study of Heart Morphogenesis Disorders in Rats in Terms of Lead Acetate Exposure at the Prenatal Stage of their Development

Shevchenko I. V.
Abstract

There are several publications dedicated to the heart morphogenesis, the articles describing the influence of various cytotoxic compounds on embryonic development are also of considerable interest. Negative effects of intoxication with lead compounds on the development of the cardiovascular system are well-known. In addition to acute toxic effects, lead accumulates in the heart and, causing pathobiochemical processes, provokes a developmental disorder. The emergence of new and modern methods of research in experimental embryology, such as electronic microscopy and immunohistochemistry, allows us to obtain new data on the cytological and morphofunctional patterns of cardiac morphogenesis. The purpose of the study was to analyze the issues of rat morphogenesis disorders under the influence of lead acetate on prenatal development and under conditions of correction of lycopene and inulin. Material and methods. Experiments were performed on Wistar rats, females (average weight 200-220 g). Animals kept the vivarium in the standard conditions: air temperature 22 ± 2° С, humidity 55 ± 15%, 12-hour light / dark cycle, free access to water and food. Intact pregnant females were given distilled water (group 1). Group 2 pregnant females were administered 2.5 g% solution of lead acetate intragastrically at a rate of 50 mg / kg of body weight per day (via a probe once daily for the entire duration of pregnancy). A separate group consisted of animals administered lead acetate and lycopene (group 3) and inulin (group 4). Aqueous solutions of lycopene (Hubei Pharmaceutical) and inulin were injected in a similar manner at a rate of 500 mg / kg of body weight per day. Results and discussion. The results of the studies showed new data on cardiac morphogenesis disorders on prenatal development with intoxication with lead compounds, a new approach to the study of normal and pathological development of the heart. The results of morphometric studies indicated a reduction in the area of nuclei of cardiomyocytes in groups administered lead acetate. Lead acetate suppresses cardiac morphogenesis in the prenatal period, which at the morphological level delays development and damage to the right and left atrium myocardium and right ventricle. The general manifestation of disturbed cardiac morphogenesis on E14-E18 was the reduction in cardiac myocardial density in the myocardium, decreasing the size of the nuclei of cardiomyocytes, increasing the interstitial space in the cortical (outer) layer and the intervals in the trabecular (internal) layer of the myocardium, decreasing the density and blood filling. Structural changes in embryos of stage E16 were delayed myocardial development, which affected the lower density of cardiomyocytes in the ventricles and atriums, the appearance of focal cells of the empty interstitial space, which may be evidence of edema. For all markers used (α-SMA, MMP-9 and VEGF), there was a finding of cardiac morphogenesis, structural and metabolic changes. Conclusion. The immunohistochemical study revealed suppression of expression of α-SMA, MMP-9 and VEGF in the myocardium within terms of 14, 16, 18 days of prenatal development, which is the manifestation of structural and functional damage to cardiomyocytes. The use of lycopene and inulin during the pregnancy of rats contributed to the restoration of the left atrium and right ventricle.

Keywords: embryogenesis, cardiogenesis, experiment, lead acetate, inulin, lycopene

Full text: PDF (Ukr) 420K

References
  1. Nefodova OO. Vyznachennya vplyvu atsetatu svyntsyu na khid kardiohenezu shchura v eksperymenti. Visnyk problem biolohiyi i medytsyny. 2014; 4(2): 243-6. [Ukrainian]
  2. Shatorna VF, Harets VI, Nefodova OO, Kononova II. Vplyv nyzkykh doz atsetatu svyntsyu na kardiohenez shchura v eksperymenti. Visnyk problem biolohiyi i medytsyny. 2016; 2(2): 375-9. [Ukrainian]
  3. Dovgal GV, Shevchenko IV. Ultrastrukturni osnovi kardiotoksichnoyi diyi acetatu svincyu na morfogenez sercya. Vistnik problem biologiyi i medicine. 2018; 2(144): 306-10. [Ukrainian]
  4. Ghosh D, Firdaus SB, Mitra E, Chattopadhyay A, Pattari SK, Dutta S, et al. Aqueous leaf extract of Murraya koenigii protects against lead-induced cardio toxicity in male wistar rats. International Journal of Phytopharmacology. 2013; 4(2): 119-32.
  5. Kumar GL, Rudbeck L. Immunogistohimicheskie metody: Rukovodstvo: DAKO. Per s angl pod red GA Franko, PG Malkov. 2011. 224 p.
  6. Hernandez-Martinez AR, Molina GA, Jimenez-Hernandez LF, Oskam AH, Fonseca G, Estevez M. Evaluation of Inulin Replacing Chitosan in a Polyurethane/Polysaccharide Material for Pb2+ Removal. Molecules. 2017 Nov 29; 22(12). pii: E2093. https://www.ncbi.nlm.nih.gov/pubmed/29186073. https://www.ncbi.nlm.nih.gov/pmc/articles/6150026. https://doi.org/10.3390/molecules22122093
  7. Kern S, Feng HZ, Wei H, Cala S, Jin JP. Up-regulation of alpha-smooth muscle actin in cardiomyocytes from non-hypertrophic and non-failing transgenic mouse hearts expressing N-terminal truncated cardiac troponin I. FEBS open bio. 2013; 4: 11-7. https://www.ncbi.nlm.nih.gov/pubmed/24319652. https://www.ncbi.nlm.nih.gov/pmc/articles/3851183. https://doi.org/10.1016/j.fob.2013.11.002
  8. Komousani TA, Moselhy SS. Modulation of lead biohazards using a combination of epicatechin and lycopene in rats. Hum Exp Toxicol. 2011 Oct; 30(10): 1674-81. https://www.ncbi.nlm.nih.gov/pubmed/21262865. https://doi.org/10.1177/0960327110396536
  9. Li N, Li X, Li L, Zhan P, Qiao M, Zhao Q, et al. Original Research. The expression of MMP2 and MMP9 in the hippocampus and cerebral cortex of newborn mice under maternal lead exposure. Experimental Biology and Medicine. 2016; 241(16): 1811–8. https://www.ncbi.nlm.nih.gov/pubmed/27190262. https://www.ncbi.nlm.nih.gov/pmc/articles/5027935. https://doi.org/10.1177/1535370216647808
  10. Liu X, Su P, Meng S, Aschner M, Cao Y, Luo W, et al. Role of matrix metalloproteinase-2/9 (MMP2/9) in lead-induced changes in an in vitro blood-brain barrier model. International Journal of Biological Sciences. 2017; 13(11): 1351–1360. https://www.ncbi.nlm.nih.gov/pubmed/29209140. https://www.ncbi.nlm.nih.gov/pmc/articles/5715519. https://doi.org/10.7150/ijbs.20670
  11. Marcela SG, Cristina RM, Angel PG, Manuel AM, Sofía DC, Patricia de LR, et al. Chronological and morphological study of heart development in the rat. Anat Rec (Hoboken). 2012 Aug; 295(8): 1267-90. https://www.ncbi.nlm.nih.gov/pubmed/22715162. https://doi.org/10.1002/ar.22508
  12. Silveira EA, Lizardo JHF, Souza LP, Stefanon I, Vassallo DV. Acute lead-induced vasoconstriction in the vascular beds of isolated perfused rat tails is endothelium-dependent. Braz J Med Biol Res. 2010 May; 43(5): 492-9. https://doi.org/10.1590/S0100-879X2010007500027
  13. Mahmoud UM, Ebied A-BM, Mohamed SM. Effect of lead on some haematological and biochemical characteristics of Clarias gariepinus dietary supplemented with lycopene and vitamin E. Egypt Acad J Biolog Sci. 2013; 5(1): 67-89.
  14. Winiarska-Mieczan A, Krusiński R, Kwiecień M. Tannic Acid influence on lead and cadmium accumulation in the hearts and lungs of rats. Adv Clin Exp Med. 2013 Sep-Oct; 22(5): 615-20. https://www.ncbi.nlm.nih.gov/pubmed/24285445