The Effect of Picroside-2 on Erythrocyte Deformability and Lipid Peroxidation in Streptozotocin-Induced Diabetic Rats Subjected to Left Anterior Descending Artery-Ischaemia Reperfusion

  • Faruk Metin Çomu
  • Yücel Polat
  • Abdullah Özer
  • Dilek Erer
  • Mehmet Kirişçi
  • Ali Doğan Dursun
  • Tolga Tatar
  • Hakan Kartal
  • Ayşegül Küçük
  • Hakan Boyunağa
  • Mustafa Arslan Gazi Üniversitesi Tıp Fakültesi
Keywords: Erythrocyte deformability, myocardial ischaemia reperfusion, experimental diabetes, picroside-2, MDA, NO


Aim: Diabetes mellitus (DM) is a chronic metabolic disorder principally characterized by an elevation in oxidative stress levels. Ischaemia-reperfusion (IR) injury starts a cascade of events that lead to tissue ischaemia and cellular damage produced by reperfusion causing an inflammatory like response.  Erythrocyte deformability and plasma viscosity are important clinical implications for organ and tissue perfusion. Recent studies have found that picroside-2 has antioxidant, neuroprotective and anti-inflamatory effects.   The aim of our study was to investigate the effects of picroside-2 on erythrocyte deformability and lipid peroxidation in streptozotocin-induced diabetic rats subjected to left anterior descending (LAD) artery IR.Methods: The animals were randomly assigned to one of five experimental groups. In Group (control) C, DC (diabetes-control group), and DP (diabetes-picroside-2 group) neither coronary artery occlusion nor reperfusion were performed in the control rats. In Group DIR, a branch of the LAD artery was occluded for 45 minutes followed by 90 minutes of reperfusion to produce IR. In Group DIRP, picroside-2 was administrated via 10 inraperitoneal (IP) route 30 minutes before ligating the LAD artery. Serum malondialdehyde and nitric oxide activities were investigated to document lipid peroxidation and erythrocyte deformability index.Results: Deformability index was notably increased in diabetic rats (p<0.0001). It was notably increased in Group DIR when compared to Group C, DC, DP and DIRP (p<0.0001, p=0.009, p=0.013, p=0.009, respectively). MDA level and NO activity were also higher in IR group than the other groups.Conclusion: Erythrocyte deformability index was decreased in rats with diabetes and IR injury. This injury may lead to further microcirculatory problems. Picroside 2 was shown to be useful in reducing the side effects of this kind of injury. 


Robertshaw HJ, Hall GM. Diabetes mellitus: anaesthetic management. Anaesthesia 2010;61: 1187-90.

McAnulty GR, Robertshaw HJ, Hall GM. Anaesthetic management of patients with diabetes mellitus. Br J Anesth 2000; 85: 80-90.

McAnulty GR, Hall GM. Anaesthesia for the diabetic patient. Br J Anesth 2003; 88: 428-30.

Gu W, Pagel PS, Warltier DC, Kersten JR. Modifying cardiovascular risks in diabetes mellitus. Anesthesiology 2003;98: 774-9.

Giquel J, Rodriguez-Blanco YF, Matadial C, Candiotti K. Diabetes mellitus in anaesthesia. Br J Diabetes Vasc Dis 2012;12: 60-4.

Miller RD. Miller’s Anesthesia. 7th ed. In: Roizen MF, Fleisher LA, eds. Anesthesia implications of current disease. Chicago: Year Book Medical Publishers, 2010:1067-149.

Kadoi Y. Anesthetic considerations in diabetic patients. Part I: preoperative considerations of patients with diabetes mellitus. J Anesth 2010; 24: 739-47.

Margolis JR, Kannel WS, Feinleib M, Dawber TR, McNamara PM. Clinical features of unrec¬ognized myocardial infarction silent and symptomatic. Eighteen year follow-up: Framingham study. Am J Cardiol 1973;32: 1-7.

Peto K, Nemeth N, Brath E, Takacs IE, Baskurt OK, Meiselman HJ, et al. The effects of renal ischemia-reperfusion on hemorheological factors: preventive role of allopurinol. Clin Hemorheol Microcirc 2007; 37: 347-58.

Vincent AM, Russell JW, Low P, Feldman EL.Oxidative stress in the pathogenesis of diabetic neuropathy. Endocrine Reviews 2004; 25: 612–28.

Memisogullari R, Taysi S, Bakan E, Capoglu I. Antioxidant status and lipid peroxidation in Type II Diabetes Mellitus. Cell Biochem Func 2003; 21: 291-6.

Sacks DB. Diabetes Mellitus. In: Burtis CA, Ashwood ER, (Ed). Tietz Texbook of clinical chemistry. Philadelphia: WB Saunders Co: 1999; pp 766-76.

Cherubini A, Ruggiero C, Polidori MC, Mecocci C. Potential markers of oxidative stress in stroke. Free Radic Biol Med 2005;39: 841-52.

Memişoğulları R, Bakan E. Levels of ceruloplasmin, transferrin, and lipid peroxidation in the serum of patients with Type 2 diabetes mellitus. J Diabetes Complications 2004;18: 193-7.

Masella R, Benedetto RD, Varý R, Filesi C, Giovannini C. Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathionerelated enzymes. J Nutr Biochem 2005;16: 577–86.

Das UN. Free radicals, cytokines and nitric oxide in cardiac failure and myocardial infarction. Mol Cell Biochem 2000;215(1-2):145-52.

Cho YI, Mooney MP, Cho DJ. Hemorheological disorders in diabetes mellitus. J Diabetes Sci Technol 2008;2: 1130-8.

Li JX, Li P, Tezuka Y, Namba T, Kadota S. Three phenyl¬ethanoid glycosides and an iridoid glycoside from Picrorhiza scrophulariiflora. Phytochemistry 1998; 48: 537 42.

Cao Y, Liu JW, Yu YJ, et al. Synergistic protective effect of picroside II and NGF on PC12 cells against oxidative stress induced by H2O2. Pharmacol Rep 2007;59: 573 9.

Smit HF, Kroes BH, van den Berg AJ, et al. Immunomodulatory and anti inflammatory activity of Picrorhiza scrophulariiflora. J Ethnopharmacol 2000; 73: 101 9.

He LJ, Liang M, Hou FF, et al. Ethanol extraction of Picrorhiza scrophulariiflora prevents renal injury in experimental diabetes via anti inflammation action. J Endocrinol 2009; 200: 347 55.

Li Q, Li Z, Xu XY, Guo YL, Du F. Neuroprotective properties of picroside II in a rat model of focal cerebral ischemia. Int J Mol Sci 2010; 11: 4580 90.

Wang L, Liu XH, Chen H, et al. Picroside II protects rat kidney against ischemia/reperfusion-induced oxidative stress and inflammation by the TLR4/NF-κB pathway. Exp Ther Med 9:1253-8.

Esterbauer H, Cheeseman KH. Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol 1990;186: 407-21.

Cortas NK, Wakid NW. Determination of inorganic nitrate in serum ana urine by a kinetic cadmium-reduction method. Clin Chem 1990; 36: 1440-3.

Le Devehat C, Khodabandehlou T, Vimeux M. Impaired hemorheological properties in diabetic patients with lower limb arterial ischaemia. Clin Hemorheol Microcirc 2001; 25: 43-8.

Girn HR, Ahilathirunayagam S, Mavor AI, Homer-Vanniasinkam S. Reperfusion syndrome: cellular mechanisms of microvascular dysfunction and potential therapeutic strategies. Vasc Endovascular Surg 2007;41: 277-93.

Biglioli P, Cannata A, Alamanni F, Naliato M, Porqueddu M, Zanobini M, Tremoli E, Parolari A. Biological effects of off-pump vs. on-pump coronary artery surgery: focus on inflammation, hemostasis and oxidative stress. Eur J Cardiothorac Surg 2003;24: 260-9.

Barnes A, Willars E. “Diabetes” in Clinical Hemorheology. Chien S, Dormandy J, Ernst E, A Matrai A, eds. Dordrecht: Martinus Nijhoff; 1987, pp 275–309.

Zinchuk VV. Erythrocyte deformability: physiological aspects. Usp Fiziol Nauk 2001; 32: 66-78.

Kuypers FA. Red cell membrane damage. J Heart Valve Dis 1998;7: 387-95.

Sivilotti ML. Oxidant stress and haemolysis of the human erythrocyte. Toxicol Rev 2004;23: 169-88.

Le Devehat C, Khodabandehlou T, Vimeux M. Relationship between hemorheological and microcirculatory abnormalities in diabetes mellitus. Diabete Metab 1994;20: 401-4.

Zimny S, Dessel F, Ehren M, Pfohl M, Schatz H. Early detection of microcirculatory impairment in diabetic patients with foot at risk. Diabetes Care 2001;24: 1810–4.

Muller R, Musikic P. Hemorheology in surgery:a review. Angiology 1987; 38: 581-92.

Mataseje A, Beder I, Kittova M, Okkelova J, Vazan R. The assessment of erythrocyte deformability by filtration rate. Bratisl Lek Listy 2003; 104:158-60.

Barnes AJ, Locke P, Scudder PR, Dormandy TL, Dormandy JA, Slack J. Is hyperviscosity a treatable component of diabetic microcirculatory disease?. Lancet 1977;2:789–91.

Original Research