Protective effects of resveratrol on permethrin-induced fetotoxicity in rats
Year 2023,
Volume: 7 Issue: 1, 21 - 28, 15.05.2023
Yasemin Yüksel
Esra Aslan
,
Murat Tosun
,
Korhan Altunbaş
,
Özlem Özden Akkaya
,
Hasan Hüseyin Demirel
,
Mehmet Bilgehan Pektaş
Abstract
Synthetic pyrethroid insecticides have been widely used for years to prevent harmful effects of insects and control disease vectors. In this study, the effects of resveratrol against the potential toxicity of permethrin, an effective pyrethroid derivative, on the fetus were investigated. Accordingly, Wistar female rats were divided into four groups as Control, Sham, Permethrin, and Permethrin + Resveratrol. Lung, liver, kidney and small intestine of developing fetuses were evaluated histopathologically. Also, Bone Morphogenetic Protein-4 (BMP-4) in bone tissue development and Fibroblast Growth Factor-1 (FGF-1) expressions in lung were examined immunohistochemically. All structures in the Control and Sham groups were normal. Permethrin caused epithelial damage, regression in bronchial and primitive alveolar development in the lung; congestion, edema and sinusoidal dilatation around the central vein in the liver; tubular epithelial degeneration, regression in glomeruli and tubule formation in the kidney; epithelial degeneration and irregularity in the villus structure in the small intestine. Immunohistochemical results indicated that permethrin administration decreased BMP-4 levels in bone tissue and FGF-1 levels in lung. Resveratrol application was found to greatly alleviate histopathological and immunohistopathological variability in all tissues. Oral consumption of permethrin by pregnant rats caused growth retardation and tissue damage in many different tissues in offspring. Intake of resveratrol during pregnancy showed protective effects against fetotoxicity caused by permethrin.
Supporting Institution
Afyon Kocatepe University Research Foundation
Project Number
(14.TIP.07).
Thanks
The authors would like to thank Afyon Kocatepe University Research Foundation for financial support (14.TIP.07).
References
- Agrawal A, Sharma B (2010). Pesticides induced oxidative stress in mammalian systems: A review. International Journal of Biological and Medical Research 1(3): 90-104.
- Aslan E, Boyacı MG, Güzel H, Pektas MB (2021). Better neuroprotective profile of caffeic acid phenyl ester over resveratrol in non-traumatic ischemia-reperfusion injury of the spinal cord. British Journal of Neurosurgery 9: 1-7.
- Bohara RA, Tabassum N, Singh MP, Gigli G, Ragusa A, Leporatti S (2022). Recent overview of resveratrol’s beneficial effects and its nano-delivery systems. Molecules 27(16): 5154.
- Borsoi FT, Neri-Numa IA, de Oliveira WQ, de Araújo FF, Pastore GM (2023). Dietary polyphenols and their relationship to the modulation of non-communicable chronic diseases and epigenetic mechanisms: A mini-review. Food Chemistry: Molecular Sciences. 6: 100155.
- Bozkurt E, Atay E, Pektas G, Ertekin A, Vurmaz A, Korkmaz OA, et al. (2020). Potential anti-tumor activity of kefir-induced juglone and resveratrol fractions against erhrlich ascites carcinoma-bearing BALB/C mice. Iranian Journal of Pharmaceutical Research 19(3): 358-369.
- Burri PH (1984). Fetal and postnatal development of the lung. Annual Review of Physiology 46(1): 617-628.
- Chen X, Zhang J, Yin N, Wele P, Li F, Dave S, et al. (2023). Resveratrol in disease prevention and health promotion: A role of the gut microbiome. Critical Reviews in Food Science and Nutrition 1(1): 1-18.
- Coleman SJ, Bruce C, Chioni AM, Kocher HM, Grose RP (2014). The ins and outs of fibroblast growth factor receptor signalling. Clinical Science 127(4): 217-231.
- Curtis GH, Nogueiro S, Schneider S, Bernhofer M, McDermott M, Nixon E, et al. (2021). Trans‐ovo permethrin exposure affects growth, brain morphology and cardiac development in quail. Environmental Toxicology 36(7): 1447-1456.
- Dekant W, Vamvakas S (1996). Biotransformation and membrane transport in nephrotoxicity. Critical Reviews in Toxicology 26(3): 309-334.
- Dhivya Vadhana MS, Nasuti C, Gabbianelli R (2010). Purine bases oxidation and repair following permethrin insecticide treatment in rat heart cells. Cardiovascular Toxicology 10(3): 199-207.
- Ding Y, Landrum PF, You J, Lydy MJ (2013). Assessing bioavailability and toxicity of permethrin and DDT in sediment using matrix solid phase microextraction. Ecotoxicology 22(1): 109-117.
- Elser BA, Hing B, Stevens HE (2022). A narrative review of converging evidence addressing developmental toxicity of pyrethroid insecticides. Critical Reviews in Toxicology 52(5): 371-388.
- Feng L, Cook B, Tsai SY, Zhou T, LaFlamme B, Evans T, et al. (2016). Discovery of a small-molecule BMP sensitizer for human embryonic stem cell differentiation. Cell Reports 15(9): 2063-2075.
- Fernandes-Freitas I, Owen BM (2015). Metabolic roles of endocrine fibroblast growth factors. Current Opinion in Pharmacology 25: 30-35.
- Gabbianelli R, Palan M, Flis DJ, Fedeli D, Nasuti C, Skarydova L, et al. (2013). Imbalance in redox system of rat liver following permethrin treatment in adolescence and neonatal age. Xenobiotica 43(12): 1103-1110.
- Gasser E, Sancar G, Downes M, Evans RM (2022). Metabolic messengers: Fibroblast growth factor 1. Nature Metabolism 4(6): 663-671.
- Gey N, Ersan Y (2020). Investigation of histopathological effects of deltamethrin on gills, liver and intestinal tissues of Carassius gibelio (Bloch, 1782). Halic University Journal of Science 3(2): 189-210.
- Gibson-Corley KN, Olivier AK, Meyerholz DK (2013). Principles for valid histopathologic scoring in research. Veterinary Pathology 50(6): 1007-1015.
- Guvenc D, Kabak Y, Atmaca E, Aksoy A, Guvenc T (2013). Examination of caspase-dependent apoptotic and necrotic changes in rat kidney exposed to different doses of permethrin. Biotechnic and Histochemistry. 88(2): 76–85.
- Jin M, Li L, Xu C, Wen Y, Zhao M (2010). Estrogenic activities of two synthetic pyrethroids and their metabolites. Journal of Environmental Sciences 22(2): 290-296.
- Koca HB, Pektas MB, Koca S, Pektas G, Sadi G. (2016). Diabetes-induced renal failure is associated with tissue inflammation and neutrophil gelatinase-associated lipocalin: Effects of resveratrol. Archives of Biological Sciences 68(4): 747-752.
- Liang YJ, Wang HP, Long DX, Li W, Wu YJ (2013). A metabonomic investigation of the effects of 60days exposure of rats to two types of pyrethroid insecticides. Chemico-Biological Interactions 206(2): 302-308.
- López-Aceves TG, Coballase-Urrutia E, Estrada-Rojo F, Vanoye-Carlo A, Carmona-Aparicio L, Hernández ME, et al. (2021). Exposure to sub-lethal doses of permethrin is associated with neurotoxicity: Changes in bioenergetics, redox markers, neuroinflammation and morphology. Toxics 9(12): 337.
- Lu G, Liu Q, Gao T, Li J, Zhang J, Chen O, et al. (2022). Resveratrol and FGF1 synergistically ameliorates doxorubicin-induced cardiotoxicity via activation of SIRT1-NRF2 pathway. Nutrients 14(19): 4017.
- Min KK, Neupane S, Adhikari N, Sohn W, An S, Kim J, et al. (2020). Effects of resveratrol on bone‐healing capacity in the mouse tooth extraction socket. Journal of Periodontal Research 55(2): 247-257.
- Mossa ATH, Mohafrash SMM, Chandrasekaran N (2018). Safety of natural insecticides: Toxic effects on experimental animals. BioMed Research International 2018: 1-17.
- Nasuti C, Fattoretti P, Carloni M, Fedeli D, Ubaldi M, Ciccocioppo R, et al. (2014). Neonatal exposure to permethrin pesticide causes lifelong fear and spatial learning deficits and alters hippocampal morphology of synapses. Journal of Neurodevelopmental Disorders 6(1): 7.
- Nguyen, QBD, Vu MA, Hebert AA (2023). Insect repellents: An updated review for the clinician. Journal of the American Academy of Dermatology 88(1): 123-130.
- Pektas A, Pektas MB, Koca HB, Tosun M, Aslan E, Koca S, et al. (2017). Effects of resveratrol on diabetes-induced vascular tissue damage and inflammation in male rats. Turkish Journal of Biochemistry 42(4): 451-458.
- Pektas MK, Pektas MB, Surmen A, Karatepe F, Akar F (2014). Can resveratrol supplementation reduce adverse effects of metabolic syndrome on ovaries? Turkiye Klinikleri Journal of Gynecology Obstetrics 24(2): 98-103.
- Pektas MB, Turan O, Ozturk G, Sumlu E, Sadi G, Akar F (2018). High glucose causes vascular dysfunction through Akt/eNOS pathway: reciprocal modulation by juglone and resveratrol. Canadian Journal of Physiology and Pharmacology. 96(8): 757–764.
- Pektas MB, Aslan E, Firat F, Karaca C, Guzel H, Yildizhan S (2021). Suppressive effects of resveratrol on epithelial-mesenchymal transition in the YKG1 glioblastoma cell-line. Medicine Science 10(4): 1293-1298.
- Roma GC, Oliveira PR, De Bechara GH, Mathias MIC (2012). Cytotoxic effects of permethrin on mouse liver and spleen cells. Microscopy Research and Technique 75(2): 229-238.
- Romero A, Ramos E, Ares I, Castellano V, Martínez M, Martínez-Larrañaga MR, et al. (2017). Oxidative stress and gene expression profiling of cell death pathways in alpha-cypermethrin-treated SH-SY5Y cells. Archives of Toxicology 91(5): 2151-2164.
- Simsek N, Altunkaynak BZ, Unal D, Can S, Malkoc I, Unal B (2009). A stereological and electron microscopic study of the development of the nephron in prenatal and postnatal rats. The Eurasian Journal of Medicine 41(2): 84-90.
- Tateiwa D, Kaito T (2023). Advances in bone regeneration with growth factors for spinal fusion: A literature review. North American Spine Society Journal 13: 100193.
- Tussellino M, Ronca R, Carotenuto R, Pallotta MM, Furia M, Capriglione T (2016). Chlorpyrifos exposure affects fgf8, sox9, and bmp4 expression required for cranial neural crest morphogenesis and chondrogenesis in Xenopus laevis embryos. Environmental and Molecular Mutagenesis 57(8): 630-640.
- Wang X, Martínez MA, Dai M, Chen D, Ares I, Romero A, et al. (2016). Permethrin-induced oxidative stress and toxicity and metabolism. A review. Environmental Research 149(1): 86-104.
- Xiao M, Tang Y, Wang AJ, Wang BJ, Lu G, Guo Y, et al. (2021). Regulatory role of endogenous and exogenous fibroblast growth factor 1 in the cardiovascular system and related diseases. Pharmacological Research 169: 105596.
- Xu X, Liu Q, Li J, Xiao M, Gao T, Zhang X, et al. (2022). Co-treatment with resveratrol and FGF1 protects against acute liver toxicity after doxorubicin treatment via the AMPK/NRF2 pathway. Frontiers in Pharmacology 13: 940406.
- Yang C, Luo P, Chen S, Deng Z, Fu X, Xu D, et al. (2021). Resveratrol sustains intestinal barrier integrity, improves antioxidant capacity, and alleviates inflammation in the jejunum of ducks exposed to acute heat stress. Poultry Science 100(11): 101459.
- Ye Y, Jiang Z, Pan Y, Yang G, Wang Y (2022). Role and mechanism of BMP4 in bone, craniofacial, and tooth development. Archives of Oral Biology 140: 105465.
- Zhang N, Xie YP, Pang L, Zang XX, Wang J, Shi D, et al. (2014). Paraquat increases connective tissue growth factor expression and impairs lung fibroblast proliferation and viscoelasticity. Human and Experimental Toxicology 33(12): 1232-1240.
Sıçanlarda permetrin kaynaklı fetotoksisite üzerine resveratrolün koruyucu etkileri
Year 2023,
Volume: 7 Issue: 1, 21 - 28, 15.05.2023
Yasemin Yüksel
Esra Aslan
,
Murat Tosun
,
Korhan Altunbaş
,
Özlem Özden Akkaya
,
Hasan Hüseyin Demirel
,
Mehmet Bilgehan Pektaş
Abstract
Sentetik piretroid insektisitler, yıllardır böceklerin zararlı etkilerinden korunmak ve hastalık vektörlerinin kontrolü için yaygın olarak kullanılmaktadır. Bu çalışmada, etkili bir piretroid türevi olan permetrinin fetüs üzerindeki olası toksisitesine karşı polifenolik bir bileşik olan resveratrolün potansiyel etkileri araştırılmıştır. Çalışmada Wistar dişi sıçanlar Kontrol, Sham, Permetrin ve Permetrin+Resveratrol olmak üzere dört gruba ayrıldı. Histopatolojik olarak fetüslerin akciğer, karaciğer, böbrek ve ince bağırsak gelişimleri değerlendirildi. Ayrıca immunohistokimyasal olarak kemik doku gelişiminde Bone Morphogenetic Protein 4 (BMP-4) ve akciğerde Fibroblast Growth Factor-1 (FGF-1) ekspresyonlarına bakıldı. Kontrol ve Sham grubunda tüm yapılar normal görünümde izlendi. Permetrin; akciğerde epitelyal hasara, bronşiyal ve primitif alveolar gelişimde gerilemeye; karaciğerde santral ven çevresinde konjesyon, ödem ve sinüzoidal dilatasyona; böbrekte tübül epitelinde dejenerasyona, glomerül ve tübül oluşumunda gerilemeye; ince bağırsakta da epitelyal dejenerasyona ve villüs yapısında düzensizliğe neden olduğu saptanmıştır. İmmünohistokimyasal sonuçlarda permetrin uygulaması ile kemik dokuda BMP-4 ve akciğerde FGF-1 ekspresyonlarında azalma saptanmıştır. Resveratrol uygulaması sonrasında tüm dokularda histopatolojik ve immünohistokimyasal değişiklikler büyük oranda hafiflemiştir. Gebe sıçanların oral yolla permetrin tüketimi, yavrularda birçok farklı dokuda gelişme geriliği ve doku hasarına neden olmaktadır. Gebelik sürecinde resveratrol alımı ise permetrinin neden olduğu fetotoksisiteye karşı koruyucu etkinlik göstermektedir.
Project Number
(14.TIP.07).
References
- Agrawal A, Sharma B (2010). Pesticides induced oxidative stress in mammalian systems: A review. International Journal of Biological and Medical Research 1(3): 90-104.
- Aslan E, Boyacı MG, Güzel H, Pektas MB (2021). Better neuroprotective profile of caffeic acid phenyl ester over resveratrol in non-traumatic ischemia-reperfusion injury of the spinal cord. British Journal of Neurosurgery 9: 1-7.
- Bohara RA, Tabassum N, Singh MP, Gigli G, Ragusa A, Leporatti S (2022). Recent overview of resveratrol’s beneficial effects and its nano-delivery systems. Molecules 27(16): 5154.
- Borsoi FT, Neri-Numa IA, de Oliveira WQ, de Araújo FF, Pastore GM (2023). Dietary polyphenols and their relationship to the modulation of non-communicable chronic diseases and epigenetic mechanisms: A mini-review. Food Chemistry: Molecular Sciences. 6: 100155.
- Bozkurt E, Atay E, Pektas G, Ertekin A, Vurmaz A, Korkmaz OA, et al. (2020). Potential anti-tumor activity of kefir-induced juglone and resveratrol fractions against erhrlich ascites carcinoma-bearing BALB/C mice. Iranian Journal of Pharmaceutical Research 19(3): 358-369.
- Burri PH (1984). Fetal and postnatal development of the lung. Annual Review of Physiology 46(1): 617-628.
- Chen X, Zhang J, Yin N, Wele P, Li F, Dave S, et al. (2023). Resveratrol in disease prevention and health promotion: A role of the gut microbiome. Critical Reviews in Food Science and Nutrition 1(1): 1-18.
- Coleman SJ, Bruce C, Chioni AM, Kocher HM, Grose RP (2014). The ins and outs of fibroblast growth factor receptor signalling. Clinical Science 127(4): 217-231.
- Curtis GH, Nogueiro S, Schneider S, Bernhofer M, McDermott M, Nixon E, et al. (2021). Trans‐ovo permethrin exposure affects growth, brain morphology and cardiac development in quail. Environmental Toxicology 36(7): 1447-1456.
- Dekant W, Vamvakas S (1996). Biotransformation and membrane transport in nephrotoxicity. Critical Reviews in Toxicology 26(3): 309-334.
- Dhivya Vadhana MS, Nasuti C, Gabbianelli R (2010). Purine bases oxidation and repair following permethrin insecticide treatment in rat heart cells. Cardiovascular Toxicology 10(3): 199-207.
- Ding Y, Landrum PF, You J, Lydy MJ (2013). Assessing bioavailability and toxicity of permethrin and DDT in sediment using matrix solid phase microextraction. Ecotoxicology 22(1): 109-117.
- Elser BA, Hing B, Stevens HE (2022). A narrative review of converging evidence addressing developmental toxicity of pyrethroid insecticides. Critical Reviews in Toxicology 52(5): 371-388.
- Feng L, Cook B, Tsai SY, Zhou T, LaFlamme B, Evans T, et al. (2016). Discovery of a small-molecule BMP sensitizer for human embryonic stem cell differentiation. Cell Reports 15(9): 2063-2075.
- Fernandes-Freitas I, Owen BM (2015). Metabolic roles of endocrine fibroblast growth factors. Current Opinion in Pharmacology 25: 30-35.
- Gabbianelli R, Palan M, Flis DJ, Fedeli D, Nasuti C, Skarydova L, et al. (2013). Imbalance in redox system of rat liver following permethrin treatment in adolescence and neonatal age. Xenobiotica 43(12): 1103-1110.
- Gasser E, Sancar G, Downes M, Evans RM (2022). Metabolic messengers: Fibroblast growth factor 1. Nature Metabolism 4(6): 663-671.
- Gey N, Ersan Y (2020). Investigation of histopathological effects of deltamethrin on gills, liver and intestinal tissues of Carassius gibelio (Bloch, 1782). Halic University Journal of Science 3(2): 189-210.
- Gibson-Corley KN, Olivier AK, Meyerholz DK (2013). Principles for valid histopathologic scoring in research. Veterinary Pathology 50(6): 1007-1015.
- Guvenc D, Kabak Y, Atmaca E, Aksoy A, Guvenc T (2013). Examination of caspase-dependent apoptotic and necrotic changes in rat kidney exposed to different doses of permethrin. Biotechnic and Histochemistry. 88(2): 76–85.
- Jin M, Li L, Xu C, Wen Y, Zhao M (2010). Estrogenic activities of two synthetic pyrethroids and their metabolites. Journal of Environmental Sciences 22(2): 290-296.
- Koca HB, Pektas MB, Koca S, Pektas G, Sadi G. (2016). Diabetes-induced renal failure is associated with tissue inflammation and neutrophil gelatinase-associated lipocalin: Effects of resveratrol. Archives of Biological Sciences 68(4): 747-752.
- Liang YJ, Wang HP, Long DX, Li W, Wu YJ (2013). A metabonomic investigation of the effects of 60days exposure of rats to two types of pyrethroid insecticides. Chemico-Biological Interactions 206(2): 302-308.
- López-Aceves TG, Coballase-Urrutia E, Estrada-Rojo F, Vanoye-Carlo A, Carmona-Aparicio L, Hernández ME, et al. (2021). Exposure to sub-lethal doses of permethrin is associated with neurotoxicity: Changes in bioenergetics, redox markers, neuroinflammation and morphology. Toxics 9(12): 337.
- Lu G, Liu Q, Gao T, Li J, Zhang J, Chen O, et al. (2022). Resveratrol and FGF1 synergistically ameliorates doxorubicin-induced cardiotoxicity via activation of SIRT1-NRF2 pathway. Nutrients 14(19): 4017.
- Min KK, Neupane S, Adhikari N, Sohn W, An S, Kim J, et al. (2020). Effects of resveratrol on bone‐healing capacity in the mouse tooth extraction socket. Journal of Periodontal Research 55(2): 247-257.
- Mossa ATH, Mohafrash SMM, Chandrasekaran N (2018). Safety of natural insecticides: Toxic effects on experimental animals. BioMed Research International 2018: 1-17.
- Nasuti C, Fattoretti P, Carloni M, Fedeli D, Ubaldi M, Ciccocioppo R, et al. (2014). Neonatal exposure to permethrin pesticide causes lifelong fear and spatial learning deficits and alters hippocampal morphology of synapses. Journal of Neurodevelopmental Disorders 6(1): 7.
- Nguyen, QBD, Vu MA, Hebert AA (2023). Insect repellents: An updated review for the clinician. Journal of the American Academy of Dermatology 88(1): 123-130.
- Pektas A, Pektas MB, Koca HB, Tosun M, Aslan E, Koca S, et al. (2017). Effects of resveratrol on diabetes-induced vascular tissue damage and inflammation in male rats. Turkish Journal of Biochemistry 42(4): 451-458.
- Pektas MK, Pektas MB, Surmen A, Karatepe F, Akar F (2014). Can resveratrol supplementation reduce adverse effects of metabolic syndrome on ovaries? Turkiye Klinikleri Journal of Gynecology Obstetrics 24(2): 98-103.
- Pektas MB, Turan O, Ozturk G, Sumlu E, Sadi G, Akar F (2018). High glucose causes vascular dysfunction through Akt/eNOS pathway: reciprocal modulation by juglone and resveratrol. Canadian Journal of Physiology and Pharmacology. 96(8): 757–764.
- Pektas MB, Aslan E, Firat F, Karaca C, Guzel H, Yildizhan S (2021). Suppressive effects of resveratrol on epithelial-mesenchymal transition in the YKG1 glioblastoma cell-line. Medicine Science 10(4): 1293-1298.
- Roma GC, Oliveira PR, De Bechara GH, Mathias MIC (2012). Cytotoxic effects of permethrin on mouse liver and spleen cells. Microscopy Research and Technique 75(2): 229-238.
- Romero A, Ramos E, Ares I, Castellano V, Martínez M, Martínez-Larrañaga MR, et al. (2017). Oxidative stress and gene expression profiling of cell death pathways in alpha-cypermethrin-treated SH-SY5Y cells. Archives of Toxicology 91(5): 2151-2164.
- Simsek N, Altunkaynak BZ, Unal D, Can S, Malkoc I, Unal B (2009). A stereological and electron microscopic study of the development of the nephron in prenatal and postnatal rats. The Eurasian Journal of Medicine 41(2): 84-90.
- Tateiwa D, Kaito T (2023). Advances in bone regeneration with growth factors for spinal fusion: A literature review. North American Spine Society Journal 13: 100193.
- Tussellino M, Ronca R, Carotenuto R, Pallotta MM, Furia M, Capriglione T (2016). Chlorpyrifos exposure affects fgf8, sox9, and bmp4 expression required for cranial neural crest morphogenesis and chondrogenesis in Xenopus laevis embryos. Environmental and Molecular Mutagenesis 57(8): 630-640.
- Wang X, Martínez MA, Dai M, Chen D, Ares I, Romero A, et al. (2016). Permethrin-induced oxidative stress and toxicity and metabolism. A review. Environmental Research 149(1): 86-104.
- Xiao M, Tang Y, Wang AJ, Wang BJ, Lu G, Guo Y, et al. (2021). Regulatory role of endogenous and exogenous fibroblast growth factor 1 in the cardiovascular system and related diseases. Pharmacological Research 169: 105596.
- Xu X, Liu Q, Li J, Xiao M, Gao T, Zhang X, et al. (2022). Co-treatment with resveratrol and FGF1 protects against acute liver toxicity after doxorubicin treatment via the AMPK/NRF2 pathway. Frontiers in Pharmacology 13: 940406.
- Yang C, Luo P, Chen S, Deng Z, Fu X, Xu D, et al. (2021). Resveratrol sustains intestinal barrier integrity, improves antioxidant capacity, and alleviates inflammation in the jejunum of ducks exposed to acute heat stress. Poultry Science 100(11): 101459.
- Ye Y, Jiang Z, Pan Y, Yang G, Wang Y (2022). Role and mechanism of BMP4 in bone, craniofacial, and tooth development. Archives of Oral Biology 140: 105465.
- Zhang N, Xie YP, Pang L, Zang XX, Wang J, Shi D, et al. (2014). Paraquat increases connective tissue growth factor expression and impairs lung fibroblast proliferation and viscoelasticity. Human and Experimental Toxicology 33(12): 1232-1240.