Mstv2_feb2008.pdf

Journal of Microscopy Society of Thailand 2008, 22: 42-45 Effect of Polysaccharide from Phellinus ignarius (L.)
Quél. on Hematological Values and Blood Cell
Characteristics in Diabetic Rats
C. Talubmook*Faculty of Environment and Resource Studies and Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham 44150, Thailand e-mail: [email protected] Abstract
The effect of polysaccharide from Phellinus ignarius (L.) Quél. (PI) on the packed cell volume, hemoglobin concentration, red and white blood cell counts, and also the blood cell characteristics was investigated in streptozotocin-induced diabetic rats. PI at a dose of 100 mg/kg b.w. was injected venously daily to the diabetic rats for seven weeks. The results showed that PI decreased the pack cell volume, hemoglobin concentration, red blood cell count, and diameter of red blood cells but not white blood cell count. In addition, PI showed some alteration of the blood cell characteristics in diabetic rats. These data indicated that long term administration of PI changed some hematological values and some characteristics in diabetic rats. Background
Diabetes mellitus is a metabolic disorder was dissolved in 2 ml normal saline before disease characterized by high blood glucose levels, intravenous injection to the diabetic rats. which result from defects in pancreatic insulin secretion responsiveness to insulin [1]. This disease is characterized by chronic hyperglycaemia and other Animals used in this study were male albino W istar rats weighting 150-220 g purchased from accompanied by symptoms of polydipsia, polyuria, the Animal Resource, National Research Council polyphagia complications affecting various organs Thailand. The rats were acclimatized in an air in the body such as the eye, kidney, nervous system and blood vessels. Hematological complications given a standard chow and watered ad libitum prior consist mainly of abnormalities in the function, to the commencing experiments. The rats were morphology and metabolism of erythrocytes, maintained in accordance with the guidelines of the leukocytes and platelets [2]. Treatment of diabetes Committee on Care and Use of Laboratory Animal mellitus and its complications in the recent context Resource, National Research Council Thailand. have focused on the usage of plant extracts [3]. The experiments performed on the rats were Antihyperglycaemic activities of plant extracts and performed in accordance with the advice of the an extract from Phellinus ignarius (L.) Quél., Institutional Animal Care and Use Committee mushroom, a member of Hymenochaetaceae, have been reported. However, the safety of extract in such treatment is still unclear whether it has any side effect(s). Moreover, the effect of the extract from PI. on hematology has not yet been The rats were injected intraperitonealy with a single dose of 65 mg/kg b.w. streptozotocin (STZ, polysaccharide from PI. on hematological values Sigma Chemicals, St. Louis, MO) freshly dissolved and blood cell characteristics in diabetic rats was in 20 mM citrate buffer adjusted pH to 4.5. After STZ injection, they were provided with a 2% sucrose solution as their drink for 48 hours to s and M ethods
alleviate the severity after initial hypoglycemic phase. Blood glucose levels were assessed three days after STZ injection to confirm the diabetic The polysaccharide from PI was kindly given stage. Rats with blood glucose level of at least 200 Phellinus Mushroom Research Center. On the day of experiment, polysaccharide at a given dosage Journal of Microscopy Society of Thailand 2008, 22: 42-45 Duncan’s New Multiple Range Test. Values of p < 0.05 were regarded as statistical significant. Group 1: Control rats received 2 ml normal saline The homological values of controls, diabetic Group 2: Diabetic rats received 2 ml normal saline controls and diabetic rats treated with PI were Group 3: Diabetic rats received 0.25 mg/kg b.w. The packed cell volume from PI treated rats was not different from diabetic control and Group 4: Diabetic rats received 100 mg/kg b.w. PI glibenclamide treated rats but was significantly The rats were administered once daily for seven (p<0.05) less than that from normal controls. The weeks. At the end of the experiment, the rats were hemoglobin concentration was also significantly anesthetized with ether. Blood samples obtained (p<0.05) less than the control, diabetic control and from the tail vein of animals were used for the glibenclamide treated rats. In addition, the red determination of hematological values. The blood blood cell count was significantly (p<0.05) less cell characteristics and ultrastructure were also than normal controls and glibenclamide treated rats. In contrast, the white blood cell count was significantly (p<0.05) higher than the controls and glibenclamide treated rats. Nevertheless, the red and white blood cell counts were not different from according with the protocol used by Chomko and Talubmook [5]. Blood sample was filled in e 1 Hematological values of controls, diabetic
heparinised capillary tube and centrifuged at 1,500 controls, diabetic rats treated with glibenclamide, and rpm for 5 min. After centrifugation, the capillary tube was placed in microhematocrit reader and the packed cell volume (PCV) was determined. The hemoglobin concentration was determined using Sahli method, hematin obtained from the reaction (x106cel
(x103cel
compared with the standard hematin. Total red and white blood cell counts were examined manually using hemocytometer after the blood was diluted. The diluting solution for red blood cell was Gower’s solution but for the white blood cell was Means + SEM within the same column followed by the different letters are significantly different at p<0.05. Abbreviations: PCV packed cell volume, Hb hemoglobin, RBC red blood cell, W BC white blood cell. e 2 Diameter of the red blood cells in of controls,
Blood smear was fixed in methanol and stained with W right-Giemsa stain for morphological study glibenclamide, and diabetic rats treated with PI.
ameter of red
Investigation of ultrastructure of blood cells was performed with the aid of the Scanning Electron Microscope. Specimens were fixed in Means + SEM within the same column followed by the different Karnovsky’s fixative, and dehydrated in an acetone letters are significantly different at p<0.05. series. Dried specimens were mounted on stubs, coated with gold and viewed in Scanning Electron Table 2 showed that the diameter of red blood cells from PI treated rats was significantly (p<0.05) smaller than those from controls, diabetic controls and glibenclamide treated rats. Figure 1 presented different characteristics of the young and old red The results of hematological values and the blood cells from controls, diabetic controls, length and width of blood cells were presented as glibenclamide treatment and diabetic rats treated the mean + SEM. Comparisons were made between with PI. The red blood cells of all rats were non- control and treatment groups using one-way nucleated biconcave disk. The old ones were small analysis of variance (ANOVA) followed by and had a prominent central pallor. However, the Journal of Microscopy Society of Thailand 2008, 22: 42-45 red blood cell characteristics from all experimental 1 Scanning electron micrographs of red blood cells
rats were not significantly different. Figure 2 from controls (A), diabetic controls (B), diabetic rats showed the smooth membrane red blood cells and treated with glibenclamide (C), and diabetic rats treated the knobby white blood cells. Nevertheless, the red and white blood cells of all experimental rats were not different. Figure 3 illustrated the white blood lymphocytes, monocytes, neutrophils, and eosinophils. Significant differences of all types of white blood cells from experimental rats were not found. 2 Scanning electron micrographs of red and white
blood cells from controls (A), diabetic controls (B), diabetic rats treated with glibenclamide (C), and diabetic rats treated with PI (D). Lymphocytes
M onocytes
Neutrophi
3 W hite blood cells from controls (A), diabetic controls (B), diabetic rats treated with glibenclamide (C), and diabetic
Journal of Microscopy Society of Thailand 2008, 22: 42-45 2. Comazzi, S., Spagnolo, V. and Bonfanti, U. In conclusion, the overall results showed that Erythrocyte changes in canine diabetes mellitus: hematological values but not ultrastructure of blood ketoacidosis. J Comp Clin Path. 2004, 12: 199- 3. Kamalakkannan, N. and Mainzen, P.S. Rutin This research was supported by grant from the streptozotocin-induced diabetic rat tissues. J development research division and Faculty of Mol Cell Biochem. 2006, 293: 211-219. Science, Mahasarakham University, Thailand. The 4. Talubmook, C., Forrest, A. and Parsons, M. author would like to thank Dr. Savittree W ongtangthintarn for her help. Thank also go to presynaptic and postsynaptic function in the rat ileum. Eur J Pharmacol. 2003, 469: 153-158. 5. Chomko, S. and Talubmook, C. Effect of leaf extracts from Morus alba and Annona squamosa References
on hematological values in diabetic rats. J Sci 1. Talubmook, C. The influence of elevated and Technol. MSU. 2007, 26: 167-173. glucose levels and the diabetic state on 6. Ponsen, S., Narkkong, N-A. and Angwanich, W . neuromuscular function in the gut, Hatfield: Morphological and ultrastructural observations University of Hertfordshire, 2002, (Ph.D. on the blood cells of Sand Lizards (Leiolepis belliana Rubritaeniata) Mertens 1961. J Anim Vet Adv. 2007, 6(4) : 522-527.

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