Role of Curcumin and Cumin on Hematological Parameters of Profenofos Exposed Mice- Mus Musculus

Swiss albino mice were exposed to profenofos (25mg/kg b.w) alternately for 30 days, thereafter the test animals were orally administered with curcumin (120mg/kg b.w) and cumin (150mg/kg b.w) to observe the medicinal effect on hematological parameters i.e., leucocytes, erythrocytes, platelets count, differential count and hemoglobin level. It was noticed that profenofos exposed mice showed considerable decrease in different blood cells and hemoglobin level. However after treatment of curcumin and cumin the levels of these blood cells were recovered. In a comparative study curcumin was found more effective than cumin. Key word: Curcumin, Cumin, Leucocytes, Erythrocytes, Platelets, Neutrophil, Hemoglobin INTRODUCTION Profenofos (O-4-bromo-2-chlorophenyl O-ethyl S-propyl phosphorothioate) is an organophosphorous pesticide and widely used to control various insects pest of agriculture/horticulture crops in developing countries . It is non biodegradable and known for its residual effect on food and drinking water. There are several earlier reports [3-8] in literature referring its adverse effects on population or wildlife which are happened to come under the exposure of profenofos . Analysis of residual quantity of profenofos in agriculture crops and in Jiv Kant Singh, Awadh Kishore Roy / Role of Curcumin... IJCPR, November 2013 January 2014, 4(4), 120-127 Pa ge 12 1 processed food community is in forefront to derive strategic measures for public health safety. Fawzy et al., (2007) [10] and Gomes et al., (1999) have also observed the hapatocellular injuries in liver and tubular degeneration of kidney respectively under the exposure to profenofos. It has also been reported that profenofos exposed animals show decrease in the level of blood cells and hemoglobin in comparison to normal range . It is also reported that some plant extract i.e., Calotropis gigantean have anti-inflamatory activity [13] and Abutilon idicum & Panax ginseng . Generation of free radicals and reactive oxygen stress (ROS) are considered as diagnostic index in profenofos poisoning. It is also reported that curcumin (1, 7-bis (4-hydroxy3methoxyphenyl) -1, 6heptadiene-3, 5 dione) have scavenging potential to free radicals and reactive oxygen stress . Seed of cumin (Cuminum cyminum) has antioxidant property . Free radicals are unpaired electrons which usually produce highly reactive free radicals. Due to intracellular reduction of O2 into ROS or free radicals, this is toxic to cells and tissues . In view of such abnormal changes in hematological parameters, the present investigation has been designed to improve blood cells count and % hemoglobin by treating profenofos exposed swiss albino mice with herbal products i.e., curcumin and cumin. MATERIALS METHODS Animals: Thirty male swiss albino mice with average body weight ranging from 22-25gm were obtained from animal house of PG Department of Zoology, Bhagalpur University, Bhagalpur, Bihar. Food and water to mice were provided ad libitum (prepared mixed formulated feed by the laboratory itself). Animals were housed in colony rooms with 12 hrs light/dark cycle at 22 ± 2 C during the period (Jan-March, 2011) of experimentation. Chemicals: Profenofos i.e., chemically O-4-bromo-2-chlorophenyl O-ethyl S-propyl phosphorothioate (50% E.C, specific gravity 1.34, trade name: "Carina", PI Industries Ltd.) was purchased from the local market. Gimsa stain, Glyerol and Ethyl alcohol were used of E. Merk Company. Treatment Protocol: Animals were placed in five groups and each group containing 6 animals, having body weight 22-25gms. GroupI represents normal control, Group-IIEthanol control, Group-IIIProfenofos exposed, Group-IV-Profenofos exposed curcumin treated and Group-V-Profenofos exposed cumin treated. Following protocols were adopted to treat each group of animals. Jiv Kant Singh, Awadh Kishore Roy / Role of Curcumin... IJCPR, November 2013 January 2014, 4(4), 120-127 Pa ge 12 2 Group-I: Normal control given normal feed for 30 days Group –II: 100μl ethanol orally administered alternately for 30 days. Group –III: animals were exposed to profenofos @ 25mg/kg b.w at alternate day till 30 days. (Profenofos was dissolved in distilled water to make a stoke solution out of which required volume was orally administered to animals to maintain does as prescribed). Group-IV: After 30 days of profenofos exposure, curcumin (120mg/kg b.w) was orally administered daily till next 30 days. Curcumin, a product of Curcuma longa was dissolved in ethanol (1mg/1ml) and required volume of it was orally administered to animals to maintain doses as prescribed. Group-V: After 30 days of profenofos exposure, cumin (150mg/kg b.w) was orally administered daily till next 30 days. Cumin seeds powder (1gm/10ml) was dissolved in distilled water for 4hrs. Collected extract was filtered through double layered muslin cloth and filtrated was further centrifuge at 5000g for 5min to get clear supernatant. Extract was concentrated in a vacuum evaporator and out of which required volume was orally administered to animals to maintain does as prescribed). Anesthesia: Animals were anesthetized by injecting intra peritoneal with a mixture ketamine (83 mg/g) and xylazine (13 mg/g) administered intra peritoneal. Hematological study: Blood sample was collected from different groups of animal’s retro orbital vein puncture for hematological study. Erythrocytes (RBCs) count, leucocytes count, platelets count differential blood corpuscle count and hemoglobin test were performed according to Schaim (1986) method. RESULTS Experimental findings of each group of animals were presented in Tables (1) & 2 and further illustrated in Figs. 1-4. The results depicted in Table -1 show that there was significant decrease in blood counts and hemoglobin in profenofos exposed group of mice where leucocytes, erythrocytes, platelets count and hemoglobin were decreased up to 70%, 56%, 15% and 44% respectively on 30 days of exposure. After 30days, different herbal treatments curcumin (120mg/kg b.w) and cumin (150mg/kg b.w) in different exposed animal groups were orally administered till 30 days and again hematological analysis was performed. It was observed that in all groups of mice blood components were noticed to achieve normal range. Platelets which Jiv Kant Singh, Awadh Kishore Roy / Role of Curcumin... IJCPR, November 2013 January 2014, 4(4), 120-127 Pa ge 12 3 play an important role in homeostasis and coagulation process in the body was found to decrease after 30days of starting treatment whereas curcumin and cumin treated animal got success in retaining its count near to normal in comparison of control group. Table: 1 Hematological analysis in different group of experimental Swiss albino mice (data represent mean value of replicates). Group of mice Total count of leucocytes (x10μl) Total count of erythrocytes (x10 μl) Platelets (x10 μl) Hemoglobin (g/dl) Normal control 20.7 12.3 940.5 14.2 Ethanol control 19.5 10.5 935.6 13.4 Profenofos 6.4 5.5 800.4 8.0 Curcumin 11.6 9.5 856.5 14.0 Cumin 11.0 9.0 840.0 13.0 Fig:1 effect of curcumin and cumin on WBC and RBC count of profenofos exposed Swiss albino mice Blood was drown from tail (n=6), cells were counted with automated counter. Values are exposed as the mean ± SEM. P<0.05 Fig:2 effect of curcumin and cumin on Platelets count of profenofos exposed Swiss albino mice Blood was drown from tail (n=6), cells were counted with automated counter. Values are exposed as the mean ± SEM.