Ghrelin effects on midgut tissue antioxidative defense and glutathione S-transferase activity in Lymantria dispar (Lepidoptera)

601 171


The aim of the study was to examine changes in Cu?Zn superoxide dismutase (SOD), catalase (CAT), their gel electrophoresis profiles, glutathione reductase (GR) activity, amount of glutathione (GSH), and the activity of glutathione S-transferase (GST) - phase II biotransformation enzyme in the midgut tissue of gypsy moth (Lymantria dispar L. (Lepidoptera: Lymantriidae)) larvae after ghrelin treatment. Four subpicomolar injections of ghrelin (0.3 pmol) or physiological saline (control) were applied every 24 h. The SOD, CAT, GR, GST activity, and amount of GSH were higher in the ghrelin-treated group than in the control. Electrophoresis gel bands of SOD and CAT had higher area and density in the treated group. The effects of ghrelin on the antioxidative defense and GST activity in insects were detected for the first time. The results provided evidence for possible application of insects as simple model systems in future studies of the role of ghrelin in the antioxidative protection of complex organisms.


Superoxide dismutase, catalase, glutathione reductase, glutathione, glutathione S-transferase

Full Text:



Aaebi H (1984). Catalase in vitro. Methods Enzymol 105: 121‒126.

Ahmad S, Pardini RS (1990). Mechanisms for regulating oxygen toxicity in phytophagous insect. Free Radical Bio Med 8: 401‒413.

Barbosa PP, Martinat P, Waldvogel M (1971). Development, fecundity and survival of the herbivore Lymantria dispar and the number of plant species in its diet. Ecol Entomol 11: 1‒6.

Beauchamp C, Fridovich I (1971). Improved assay and an assay applied to acrylamide gels. Analyt Biochem 44: 276‒284.

Beutler E (1982). Catalase. In: Beutler E, editor. Red Cell Metabolism. A Manual of Biochemical Methods. 3rd ed. New York, NY, USA: Grune and Stratton, pp. 105‒106.

Bi JL, Felton GW (1995). Foliar oxidative stress and insect herbivory: primary compounds, secondary metabolites, and reactive oxygen species as components of induced resistance. J Chem Ecol 21: 1511‒1530.

Davis BJ (1964). Disc electrophoresis method and application to human serum proteins. Ann NY Acad Sci 121: 404‒427.

Dornonville de la Cour C, Björkqvist M, Sandvik AK, Bakke I, Zhao CM, Chen D, Håkanson R (2001). A-like cells in the rat stomach contain ghrelin and do not operate under gastrin control. Regul Pept 99: 141‒150.

Felton GW, Summers CB (1995). Antioxidant systems in insects. Arch Insect Physi Biochem 29: 187‒197.

Glatzle D, Vuilleumier JP, Weber F, Decker K (1974). Glutathione reductase test with whole blood, a convenient procedure for the assessment of the riboflavin status in humans. Experientia 30: 665‒667.

Griffith OW (1980). Determination of glutathione and glutathione disulfide using glutathione reductase and 2 vinyl pyridine. Anal Biochem 106: 207‒212.

Habig WH, Pabst MJ, Jakoby WB (1974). Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 249: 7130‒7139.

Halliwell B, Gutteridge JMC (2007). Free Radicals in Biology and Medicine. 4th ed. New York, NY, USA: Oxford University Press.

Ilijin L, Vlahović M, Perić-Mataruga V, Kmetič I, Gavrilović A, Matić D, Mrdaković M. (2014). Temperature-induced stress response in Lymantria dispar neurosecretory neurons. Turk J Biol 38: 157‒167.

Kheradmand A, Alirezaei M, Birjandi M (2010). Ghrelin promotes antioxidant enzyme activity and reduces lipid peroxidation in the rat ovary. Regul Pept 162: 84‒89.

Kojima M, Ida T, Sato T (2008). Structure of mammalian and nonmammalian ghrelins. Vitam Horm 77: 31‒46.

Mathews MC, Summers CB, Felton GW (1997). Ascorbate peroxidase: a novel antioxidant enzyme in insects. Arch Insect Biochem Pharm 34: 57‒68.

Misra HP, Fridovich I (1972). The role of superoxide anion in the antioxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247: 3170‒3175.

Mittapalli O, Neal JJ, Shukle RH (2007). Antioxidant defense response in galling insect. PNAS 6: 1889‒1894.

Mrdaković M (2012). Ghrelin effect on nutritional indices, midgut and fat body of Lymantria dispar L. (Lymantriidae). Peptides 37: 55‒62.

Mrdaković M, Perić-Mataruga V, Ilijin L, Vlahović M, Janković- Tomanić M, Mirčić D, Lazarević J (2013). Response of Lymantria dispar (Lepidoptera: Lymantriidae) larvae from differently adapted populations to allelochemical stress: effects of tannic acid. Eur J Entomol 110: 55‒63.

O’Dell TM, Butt CA, Bridgeforth AW (1984). Lymantria dispar. In: Singh P, Moore R, editors. Handbook of Insect Rearing. Vol. 2. New York, NY, USA: Elsevier, pp. 355‒367.

Perić-Mataruga V, Blagojević D, Lazarević J, Spasić M (2000). Host plant effects on the presence of superoxide dismutase isoforms in gypsy moth midgut. Biologia 55: 525‒531.

Perić-Mataruga V, Blagojević D, Spasić MB, Ivanović J, Janković- Hladni M (1997). Effect of the host plant on the antioxidative defence in the midgut of Lymantria dispar L. caterpillars of different population origins. J Insect Physiol 43: 101‒106.

Perić-Mataruga V, Janać B, Vlahović M, Mrdaković M, Ilijin L, Matić D, Milošević V (2012a). Ghrelin effects on the activities of digestive enzymes and growth of Lymantria dispar L. Arch Biol Sci 64: 497‒502.

Perić-Mataruga V, Lazarević J, Blagojević D, Pavlović S (2003). Response of the antioxidative defence to flavonoid quercetin in two populations of Lymantria dispar L. In: Proceedings of the 3rd International Plant Protection Symposium. International Association for the Plant Protective Sciences, pp. 252‒260.

Perić-Mataruga V, Lazarević J, Vlahović M, Mrdaković M, Ilijin L (2006). Histology of the midgut and peritrophic membrane in Lymantria dispar caterpillars fed on leaves of Quercus cerris or Robinia pseudoaccacia. Phytoparasitica 34: 49‒53.

Perić-Mataruga V, Mirčić D, Vlahović M, Mrdaković M, Todorović D, Stevanović D, Milošević V (2009). Effects of ghrelin on the feeding behavior of Lymantria dispar L. (Lymantriidae) caterpillars. Appetite 531: 147‒150.

Perić-Mataruga V, Vlahović M, Janać B, Ilijin L, Janković-Tomanić M, Matić D, Mrdaković M (2012b). Ghrelin effect on nutritional indices, midgut and fat body of Lymantria dispar L. (Lymantriidae). Peptides 37: 55‒62.

Perić-Mataruga V, Vlahović M, Mrdaković M, Todorović D, Matić D, Gavrilović A, Ilijin L. (2014). Prothoracicotropic hormone- producing neurosecretory neurons and antioxidative defense in midgut of Lymantria dispar in trophic stress. Turk J Biol 38: 403‒411.

Salin ML, McCord JM (1974). Superoxide dismutase in polymorphonuclear leukocytes. J Clin Invest 54: 1005‒1009.

Zwirska-Korczala K, Adamszyk-Sowa M, Sowa P, Pilc K, Suchanek R, Pierzchala K, Namyslowski G, Misiolek M, Sodowski K, Kato I et al. (2007). Role of leptin, ghrelin, angiotensin II and orexins in 3T3 L1 preadipocyte cells proliferation and oxidative metabolism. J Physiol Pharmacol 58: 53‒64.