|Year : 2019 | Volume
| Issue : 1 | Page : 8-19
Experimental study on effect of Naga Bhasma (lead calx) on hematological and biochemical parameters
Dhirajsingh S Rajput1, Nilesh Mahakal2, Biswajyoi J Patgiri3
1 Department of Rasshastra and Bhaishajya Kalpana, Mahatma Gandhi Ayurved College, Hospital and Research Centre, Wardha, Maharashtra, India
2 Department of Rasashastra and Bhaishajya Kalpana, Shri Gurudeo Ayurveda College, Amravati, Maharashtra, India
3 Department of Rasashastra and Bhaishajya Kalpana, Institute for Post Graduate Training and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat, India
|Date of Web Publication||27-Aug-2019|
Dr. Dhirajsingh S Rajput
Department of Rasashastra and Bhaishajya Kalpana, Mahatma Gandhi Ayurved College Hospital and Research Centre, Salod (H), Wardha 442001, Maharashtra.
Source of Support: None, Conflict of Interest: None
Background: In recent decade, a lot of concerns has been raised regarding heavy metal content in Ayurveda medicines, especially Bhasma (metallic calx). Naga Bhasma (lead calx) is of the metallic preparation primarily indicated in treating chronic ailments such as Kamala (jaundice), Vatarakta (gout), Grdhrasi (sciatica), Kustha (skin diseases), and Prameha (diabetes). As being a compound of lead, its chronic toxicity study was planned to find out the effect of Naga Bhasma on serum biochemical and hematological parameters. Material and method: Naga Bhasma was prepared by Parada (mercury) and Gandhaka media (NBP) and Vasa Swarasa (juice of Adhatoda vasica Linn.) as herbal media (NBH) in seven Puta (incineration cycles). Chronic oral toxicity study was carried out as per Organization for Economic cooperation and Development (OECD) 408 (90-day oral repeated dose toxicity study). Wistar strain albino rats (Rattus norvegicus) were used for the experimentation. The study was conducted at three dosage levels, therapeutically effective dose (TED) of NBP and NBH was 45 mg/kg body weight of rat, TED × 5 and TED × 10. On 91st day, blood was collected by supraorbital puncture with the help of microcapillary tubes under mild ether anesthesia for estimation of serum biochemical and hematological parameters. Observation and results: Among hematological parameters, significant decrease was observed in MCV (Mean corpuscular volume), MCH (Mean corpuscular haemoglobin), and mean corpuscular hemoglobin concentration by NBH-treated group, whereas in biochemical parameters, both test drugs significantly increased BSL (Blood Sugar Level), SGOT (Serum Glutamic-Oxaloacetic Transaminase), and serum ALP (Alkaline Phosphatise) activity. Hematological and biochemical analysis indicates that both test drugs (prepared in seven incineration cycles) were not found completely safe, especially at higher dosage where the drug affected renal and hepatic functions. Conclusion: For the safety purpose, it is suggested that Naga Bhasma prepared in less number of incineration cycles may not be used for therapeutic purpose.
Keywords: Chronic toxicity, hematological and biochemical analysis, Naga Bhasma, seven Puta
|How to cite this article:|
Rajput DS, Mahakal N, Patgiri BJ. Experimental study on effect of Naga Bhasma (lead calx) on hematological and biochemical parameters. J Indian Sys Medicine 2019;7:8-19
|How to cite this URL:|
Rajput DS, Mahakal N, Patgiri BJ. Experimental study on effect of Naga Bhasma (lead calx) on hematological and biochemical parameters. J Indian Sys Medicine [serial online] 2019 [cited 2020 Jun 6];7:8-19. Available from: http://www.joinsysmed.com/text.asp?2019/7/1/8/265523
| Introduction|| |
Naga Bhasma (lead calx), which is an Ayurvedic hero-metallic preparation, is obtained from metallic lead by repeated levigation with herbal juice followed by incineration cycles. According to classical text of Ayurveda, Naga Bhasma is among the preparations possessing miraculous properties and it is indicated in treating diseases such as diarrhea, spleen enlargement, and diabetes. However, for a metallic preparation, if the utilized media for processing and the number of Puta (incineration cycles) are different then the property of final product will also be different. In context of Naga Bhasma, there are 97 methods of Bhasma preparation ranging from 1 to 100 incineration cycles. Among these methods, research was undertaken on two procedures, namely Naga Bhasma prepared by Parada (mercury) and Gandhaka media (NBP) and Vasa Swarasa (juice of Adhatoda vasica Linn.) media (NBH) in seven incineration cycles. These two procedures were chosen because these two media are the most common in 97 methods of Naga Bhasma preparation. Comparative chronic toxicity study was conducted to access the effect of Naga Bhasma on hematological, biochemical parameters, and vital organs of Wistar albino rats. As hematological and biochemical parameters represent the normal functioning of vital organs in all living beings, therefore in this paper, attempt has been made to highlight effects observed on hematological and biochemical parameters.
| Materials and Methods|| |
Animals: Wistar strain albino rats (Rattus norvegicus) were used for the experimentation. The rats were obtained from animal house attached to Institute for Postgraduate Training and Research in Ayurveda (IPGT and RA), Gujarat Ayurveda University, Jamnagar, Gujarat, India. The experimental protocols were approved by the Institutional Animal Ethics Committee (IAEC/10/2012/13) in accordance with the guideline formulated by Committee for the Purpose of Control and Supervision of Experiments on Animals, India. All the selected animals were kept under acclimatization for 15 days before experimentation. The animals were marked with saturated picric acid for proper identification.
Preparation of test formulations for administration: Naga Bhasma was prepared by two different methods, that is, NBP and by NBH in the Department of Rasa Shastra and Bhaishajya Kalpana, IPGT and RA, Gujarat Ayurveda University, Jamnagar, Gujarat, India.
Dose fixation: Dose of the drug was calculated by extrapolating the human therapeutic dose to rat on the basis of body surface area ratio (conversion factor 0.018 for rat and 0.0026 for mice) by referring to the table of Paget and Barnes (1964). The dose of both samples of Naga Bhasma is 2 Ratti according to classics of Rasashashtra. In this study, human dose of both the Bhasmas has been decided as 2 Ratti, that is, 250 mg per day.
Naga Bhasma was administered along with Sahapana, the proportion is given as follows:
250 mg Naga Bhasma + 125 mg Amalakichurna + 125 mg Haridrachurna
Total dose of Naga Bhasma with Sahapana (vehicle) = 500mg/day, adult human dose.
Considering adult human dose of both the Bhasmas, the therapeutic dose for experimental study was calculated as follows:
Rat dose = Adult human dose × body surface area ratio convertible factor.
= 500 mg × 0.018
= 9mg/200g of rat = 45mg/kg body weight of rat
Route of drug administration: The test drugs were administered through oral route at different dose levels to respective groups. Suspension of Bhasma samples along with Sahapana was prepared in honey and distilled water as per dose level and the same was administered in a dose of 5mL/kg body weight.
Drug schedule: The test drug and the vehicle were administered between 7:30 am to 8:30 am, daily.
Grouping and posology: Chronic oral toxicity study was carried out as per OECD 408 (90-day oral repeated dose toxicity study). Wistar strain albino rats of either sex weighing 200 ± 20g were selected in the experimentation and divided randomly into 10 groups, each containing six animals, three males and three females as given in [Table 1].
The test drug was made into fine suspension in honey with suitable concentration. It was administered orally once a day for 90 consecutive days. All the animals were dosed with constant dose volume (5mL/kg body wt). Body weight was noted down before commencement of the study and afterward every 7th day along with general behavior pattern by exposing each animal to open arena. On 90th day, all animals of Groups I–VII were kept for overnight fasting. Next day, blood was collected by supraorbital puncture with the help of microcapillary tubes under mild ether anesthesia for the estimation of serum biochemical and hematological parameters. Additional six animals were kept in satellite control (Group VIII) and in the therapeutically effective dose (TED) × 10 treated group (Group IX and X) for observation, after the treatment period, of reversibility or persistence of any toxic effects. The duration of this posttreatment period was fixed as 30 days.
Hematological parameters tested for chronic toxicity: Total white blood cell (TWBC) count, differential count (neutrophils, lymphocytes, eosinophils, monocytes, and basophils), hemoglobin, packed cell volume (PCV), erythrocyte sedimentation rate, red blood cell (RBC) count, platelet count, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC).
Biochemical parameters tested for chronic toxicity: The biochemical parameters were serum urea (urease–glutamate dehydrogenase–fixed time, kinetic, enzymatic method), creatinine (modified Jaffe’s reaction), total protein (Biuret method, end method), albumin (bromocresol green dye method, end point), alkaline phosphatase (International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) method, kinetic method), serum glutamic-oxaloacetic transaminase (SGOT) (IFCC method without pyridoxal phosphate), and serum glutamic pyruvic transaminase (SGPT) activity (IFCC method, kinetic without pyridoxal phosphate), total bilirubin, direct bilirubin, total protein, albumin, globulin, uric acid, and serum calcium.
| Observation and Results|| |
Effect on behavioral changes: No behavioral changes were observed in both treated groups during course of chronic toxicity study. No mortality was observed in any of the groups at TED × 10, TED × 5, and TED dose level.
Effect on food and water consumption, fecal, and urine output: A moderate increase in food intake was observed in both test drug treated groups. Fecal and urine output remained unaffected in both the groups.
Effect on body weight: Constant body weight was observed in all groups during both main study (90 days) as well as recovery study (120 days). NBP at TED and TED × 5 dose level produced significant gain in body weight as compared to initial values. NBH at all dose levels produced significant increase in body weight of rats during 90 days of toxicity study. In recovery study, during the main course of study, not much increase in body weight was observed but after discontinuation of both drugs there was a drastic increase in body weight of rats in the recovery phase. However, NBP group showed significant difference, whereas NBH drug did produce nonsignificant increase in body weight [Tables 2] and .
|Table 2: Effect of NBP and NBH on body weight of albino rats in chronic toxicity study|
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|Table 3: Effect of NBP and NBH on body weight of albino rats in chronic toxicity study|
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Effect on hematological parameters: Effect on TWBC and neutrophil in chronic toxicity study of NBP and NBH has been depicted in [Table 4]. Significant decrease in TWBC was observed at TED × 10 dose level in NBH-treated group. All groups showed statistically nonsignificant decrease in TWBC except at TED and TED × 5 dose level in NBP- and NBH-treated groups, respectively. Recovery study also showed nonsignificant increase in TWBC content in comparison to recovery control group. Maximum increase in neutrophil (14.06%) was detected in NBH at TED dose level, whereas NBH at higher dose level produced nonsignificant decrease in neutrophil percentage. NBP at all dosage levels did not produce any changes in neutrophil percentage in comparison to control group. In recovery study, nonsignificant decrease in neutrophil was seen in NBP-treated group in comparison to that in recovery control group.
|Table 4: Effect on total WBC and neutrophil in chronic toxicity study of NBP and NBH|
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[Table 5] shows the effect on lymphocyte and eosinophil in chronic toxicity study of NBP and NBH. NBP and NBH did not affect the lymphocyte percentage even after chronic administration in rats in comparison to respective group except NBH at therapeutic dose level produced nonsignificant decrease in lymphocyte content. The eosinophil counts in treated groups were almost same as that seen in the respective control group, and observed marginal changes were statically nonsignificant.
|Table 5: Effect on lymphocyte and eosinophil in chronic toxicity study of NBP and NBH|
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Effect on monocyte and Haemoglobin (Hb) percentage in chronic toxicity study of NBP and NBH is given in [Table 6]. No significant changes were observed in monocyte at all three dosage levels and in recovery study of both the groups. NBP at TED × 10 and NBH at TED showed 16.5% increase in monocyte, respectively, whereas NBH at higher dose level showed 16.5% decrease in monocytes. Statistically significant increase in hemoglobin content was observed in NBP-treated group at TED × 10 dose in comparison to control group. NBP and NBH at other dose level did not affect hemoglobin content in rats. However, in recovery study, both drugs significantly reduced the hemoglobin in comparison to recovery control group.
|Table 6: Effect on monocyte and Hb% in chronic toxicity study of NBP and NBH|
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[Table 7] indicates the effect of NBP and NBH on PCV and total RBC in chronic toxicity. Significant increase in PCV percentage was detected in NBP at TED × 10 dose level. NBP and NBH at other dose level did not affect the PCV% in comparison to control group in main study, whereas both drugs in recovery study produced nonsignificant decrease in PCV% in comparison to recovery control group. Total RBC content was nonsignificantly decreased in higher dose of NBP and NBH recovery groups. At all other dose levels, slight increase in total RBC was seen but these changes were not significant.
|Table 7: Effect on PCV and total RBC in chronic toxicity study of NBP and NBH|
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Effect on platelets and MCV in chronic toxicity study of NBP and NBH is shown in [Table 8]. Nonsignificant decrease in platelets was observed at all three dose levels in NBP, whereas NBH showed nonsignificant increase in comparison to control group. In recovery study, both the groups showed increase in platelets however this increase was significant only in NBH-treated group. NBP at TED dose, NBH at TED and TED × 10 dose level, and in NBH recovery study, significant decrease in MCV was observed in comparison to respective control group.
|Table 8: Effect on platelets and MCV in chronic toxicity study of NBP and NBH|
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[Table 9] shows the effect on MCH and MCHC in chronic toxicity study of NBP and NBH. NBP-treated group did not produce any effect on MCH and MCHC in comparison to control group. NBH at TED × 5 and TED × 10 dose level produced significant decrease in MCH and MCHC in comparison to control group. In recovery study, NBH at TED × 10 dose level produced significant decrease in MCH in comparison to recovery control group.
|Table 9: Effect on MCH and MCHC in chronic toxicity study of NBP and NBH|
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Effect on biochemical parameters: Effect on blood glucose and cholesterol level in chronic toxicity study of NBP and NBH is given in [Table 10]. Statistically significant increase in FBS was detected at all three dosage levels in both groups except TED × 10 dose in NBH. This increase was the highest in NBH (47.98%) at TED dose level. All groups showed nonsignificant changes in serum cholesterol at all three dosage levels. NBP and NBH at higher dose level produced 10.19% and 12.73% increase in serum cholesterol level in comparison to control group. In recovery study, NBP showed decrease, whereas NBH showed increase in serum cholesterol; however, both changes were nonsignificant in comparison to vehicle control group.
|Table 10: Effect on blood glucose and cholesterol level in chronic toxicity study of NBP and NBH|
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[Table 11] indicated the effect on serum triglycerides and high-density lipoprotein (HDL) cholesterol in chronic toxicity study of NBP and NBH. NBP at TED and TED × 10 levels and NBH at all dosage levels produced statistically nonsignificant decrease in serum triglycerides in comparison to control group. Significant decrease in HDL cholesterol was seen in NBP at TED × 5 and NBH at TED dose level in comparison to that in control group. Nonsignificant decrease in HDL cholesterol was seen in both groups at all remaining dose levels except at TED × 10. In recovery study, NBP produced nonsignificant decrease, whereas NBH produced nonsignificant increase in serum triglyceride and HDL-cholesterol level in comparison to recovery control group.
|Table 11: Effect on serum triglycerides and HDL cholesterol in chronic toxicity study of NBP and NBH|
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[Table 12] shows the effect on urea and serum creatinine in chronic toxicity study of NBP and NBH. Administration of NBP resulted in slight increase in blood urea at TED and TED × 10 dose, whereas at TED × 5 dose level, decrease in blood urea was detected. NBH at all dosage levels produced nonsignificant decrease in urea level in comparison to control group. In recovery study, there was a decrease in urea level in both treated group; however, NBH only showed statistically significant effect in comparison to recovery control group. Significant increase in serum creatinine level was seen at TED dose in NBP, whereas NBH showed significant decrease at TED × 5 dose. Both drugs, in recovery, showed nonsignificant decrease in serum creatinine level in comparison to recovery control group.
|Table 12: Effect on blood urea and serum creatinine in chronic toxicity study of NBP and NBH|
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[Table 13] shows the effect on SGPT and SGOT in chronic toxicity study of NBP and NBH. Administration of NBP and NBH resulted in nonsignificant increase in SGPT at all dosage levels including recovery study. Maximum of 19.25% and 28.87% was observed in NBP and NBH at higher dose level. Chronic administration of NBP and NBH for 90 days resulted in marked elevation of SGOT level in serum of treated rats. Test drugs at all dosage levels produced significant increase in SGOT level in comparison to control group. Even after discontinuation of drugs in recovery study, animals showed higher level of SGOT in serum in comparison to recovery control group.
|Table 13: Effect on SGPT and SGOT in chronic toxicity study of NBP and NBH|
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Effect on total protein and albumin in chronic toxicity study of NBP and NBH is shown in [Table 14]. Total protein was observed to be significantly increased in NBP at TED dose, whereas in TED × 5 and TED × 10 dose-treated group, it was significantly decreased in comparison to control group. NBH did not show any effect on protein content in chronic toxicity study. Both groups showed nonsignificant effect on albumin level in comparison to control group.
|Table 14: Effect on total protein and albumin in chronic toxicity study of NBP and NBH|
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[Table 15] shows effect on globulin and alkaline phosphate in chronic toxicity study of NBP and NBH. Administration of NBP resulted in decreasing globulin at all three dosage levels and in NBH at TED × 10 dose, whereas NBH showed increase at TED and TED × 5 dose. Both recovery groups showed increase in globulin but all these changes were statistically nonsignificant. Chronic administration of NBP and NBH for 90 days resulted in marked elevation of alkaline phosphatase level in serum of treated rats. Test drugs at all dosage levels produced significant increase in ALP level in comparison to control group. Even after discontinuation of drugs in recovery study, animals showed higher level of ALP in serum in comparison to recovery control group.
|Table 15: Effect on serum globulin and alkaline phosphate in chronic toxicity study of NBP and NBH|
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Effect on total bilirubin and direct bilirubin in chronic toxicity study of NBP and NBH is given in [Table 16]. Increase in total bilirubin was detected at TED and TED × 5 dose in NBH-treated group, whereas decrease was observed in remaining dose levels in both groups including recovery study; however, all these changes were statistically nonsignificant. D bilirubin also showed nonsignificant decrease at TED, TED × 5 in NBP and in recovery study. At remaining dose levels, D bilirubin was observed to be same as that of control group.
|Table 16: Effect on total bilirubin and direct bilirubin in chronic toxicity study of NBP and NBH|
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[Table 17] presents the effect on uric acid and serum calcium in chronic toxicity study of NBP and NBH. Both groups at all dosage levels and in recovery study showed nonsignificant decrease in uric acid level in comparison to control group. Statistically nonsignificant increase in serum calcium level was seen at all dosage levels in both groups except at TED dose in NBP in comparison to control group. However, serum calcium was found to be significantly increased in recovery study of both test drugs in comparison to recovery control group.
|Table 17: Effect on uric acid and serum calcium in chronic toxicity study of NBP and NBH|
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Effect on serum low-density lipoprotein (LDL) and very low-density lipoproteins (VLDL) in chronic toxicity study of NBP and NBH is shown in [Table 18]. Administration of NBP and NBH at all three dosage levels resulted in nonsignificant decrease in LDL except at TED × 10 dose in NBH-treated group, which showed nonsignificant increase. Similarly, nonsignificant decrease was observed in VLDL in both treated groups in comparison to control group. In recovery study, NBP showed increase in VLDL in comparison to recovery control group.
|Table 18: Effect on serum LDL and VLDL in chronic toxicity study of NBP and NBH|
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| Discussion|| |
Effect on hematological parameters: Administration of both test drugs resulted in significant changes in five parameters; they are significant increase in Hb percentage and packed cell volume of NBP group at 10 TED dose level, significant decrease in total WBC at 10 TED dose level in NBH, statistically significant decrease in MCV observed in NBP at TED dose level and in NBH at TED and TED × 10 dose level, and significant decrease in MCHC in NBH-treated group at TED 5 and TED 10 dose level.
The hemoglobin level showed significant increase in NBP at higher dose level. The increase in hemoglobin content in NBP-treated group may be the result of increase in the number of circulating red cells, packed cell volume, their concentration of hemoglobin, or any combination of these. In this study, it may be suggested that the increase in hemoglobin content is due to the increase observed in the production of erythrocytes in NBP-treated group. MCHC is the average concentration of hemoglobin in RBCs. MCHC is used to help diagnose the type (cause) and severity of anemia. When MCHC is low, this can mean a person has iron-deficiency anemia. This type of anemia can be caused by insufficient iron in the diet or by blood loss. Blood loss, such as those that might occur with tumors in the colon and other parts of gastrointestinal tract, can cause low iron levels and a low MCHC. The most common causes of decrease in MCV are iron deficiency, gastrointestinal blood loss or menstrual blood loss, or chronic disease. NBH at higher dose level significantly affects the mean cell volume, concentration of hemoglobin but same was not reflected in the level of TRBC and hemoglobin level, hence the observed effect was not classified as serious toxic effect of NBH on RBC-related parameters. However, NBH-treated group showed statistically significant increase in platelet content and significant decrease in MCH and MCHC.
Lead inhibits porphobilinogen synthase and ferrochelatase, preventing both porphobilinogen formation and the incorporation of iron into protoporphyrin IX, the final step in heme synthesis. Lead can also impair the activity of pyrimidine 5’-nucleotidase, increasing the pyrimidine nucleotides in RBCs and preventing the maturation of erythroid elements, which leads to decreased RBC counts and eventually anemia. This causes ineffective heme synthesis and subsequent microcytic anemia. However, hemoglobin levels do not start to decrease as a result of lead exposure until blood lead levels are 50 μg/dL. This indicates, up to recovery study blood lead level reached 50 μg/dL and resulted in significant decrease in Hb percentage. Further decrease in hemoglobin content in both the treated groups may be due to the result of diminution in the number of circulating RBCs as observed in both treated groups. In addition, significant decrease in the size of RBCs and their concentration of hemoglobin in NBH-treated group may also be responsible for decrease in hemoglobin level.
The number of circulating WBCs decreased in variety of disorders: Leukopenia that results from reduced number of neutrophils and lymphopenia, which is less common. In addition, it is observed to with cytotoxic drugs, malnutrition and autoimmune disorders or suppression of hemopoietic stem cells. The most common cause of leucopenia is drug toxicity because such drug caused a generalized suppression of bone marrow, production of RBC, and platelets were also affected. In this study, other circulating cells were not affected at significant level, hence it was suggested that drugs may affect the bone marrow production but may deplete the WBC from systematic circulation. Normally, leukocytosis occurs in association with acute inflammatory reactions, tissue necrosis, thrombosis, hemorrhage, acute lysis of red cells, and sometimes in neoplastic diseases. In the recovery study, nonsignificant increase in WBC count was observed in both treated groups, which was reversal of adverse effects of drugs observed in the main study. The observed values were within the normal range.
Effect on serum biochemical parameters: Administration of both test drugs resulted in significant increase in BSL at all dosage levels and even in recovery study. The observed hyperglycemia in this study may be due to high turnover of carbohydrate material or may be due to impaired transport of glucose and lowered glomerular filtration rate. The effect has been correlated with the adverse changes in histopathological study of kidney. Creatinine blood level depends on its production and excretion. The significant increased level of creatinine in NBP at TED dose level is still within normal range in this study. Further, the same was decreased at other dose level, which indicates that drugs may not produce any adverse effect on creatinine level.
Serum SGOT and SGPT level: SGOT is found in the liver, heart (cardiac muscle), skeletal muscle, kidneys, brain, and RBCs. In this study, significant increase in SGOT in all treated groups indicated the adverse effect of NBP and NBH on heart, kidney, and liver. In both the groups, there was a significant elevation in serum GPT levels. This enzyme is found in most of the tissues but the amount varies. The liver contains highest amount of SGPT in comparison to other tissues. Elevation of serum GPT is indicative of liver injury due to leakage of this enzyme from the tissue into the serum. Increased activity is observed in inflammatory, degenerative, and neoplastic lesions of the liver. Corroborative evidences could also be observed in histopathological studies of liver, kidney, and heart in both treated groups. This may be indicative of a toxic effect of drugs in rats in main study and even after discontinuation of drugs in recovery study.
Serum urea: Urea is the main product of protein metabolism in the body. The kidney plays important role in balancing the urea level. The increase in urea content in NBP-treated group at both the high dose level indicates impairment in the functioning of the kidney. However, the values did not reach at significant level. NBH did not affect the urea level. After discontinuation of drug, there was decrease in the urea content in comparison to normal control rats.
Serum alkaline phosphatase: This enzyme is found in most of the tissues. However, the osteoblasts in the bone, bile canaliculi in liver, epithelia cells in the intestine, proximal tubules of the kidney, placenta, and lactating mammary glands are the richest sources. In this study, significant increase in the level of this enzyme was observed at all the dosage levels of NBP and NBH and even in recovery groups. The damage to the kidney, intestine, and liver as observed in the histopathological studies in this study may be responsible for increase in ALP level.
Serum bilirubin: Bilirubin is mainly produced by the breaking down of hemoglobin in senescent RBCs and from the breakdown of other heme-containing proteins. In this study, the bilirubin was not affected at significant level at higher dose group and values observed were still within the normal range.
Effect on HDL and LDL cholesterol levels: Lipid metabolism to a great extent depends on the formation and turnover of lipoproteins. Almost all lipids in the plasma are transported in the form of complexes with proteins; these proteins are termed as lipoproteins. They are particles with a core region containing cholesteryl esters and triglycerides. This core is surrounded by a monolayer of unesterified cholesterol and phospholipid. Protein specific to the lipoproteins known as apolipoproteins are located on the outer surface of the lipoprotein molecule. Chylomicrons are the largest of the lipoproteins; they are formed in the intestine and carry triglyceride in dietary region. Triglyceride is removed from them at extrahepatic sites through hydrolysis caused by the activity of the enzyme lipoprotein lipase. After the triglyceride depletion, the remaining surface lipids and protein components are transferred to HDL moiety and the remaining chylomicron remnants are taken up by the hepatocytes. VLDL is secreted by the liver. The lipid portion of HDL comes from the surface layer of chylomicrons and VLDL, when they are hydrolyzed. It also takes up cholesterol from the cells.
In this study, nonsignificant increase in cholesterol level and decrease in HDL-cholesterol level suggested that drugs may have effect on cholesterol turnover in the body. However, significant effects were not seen in the aforementioned parameters. Further triglyceride level and LDL-cholesterol level were not affected by the drugs, hence it can be suggested that the observed increase did not show any serious toxic effect. Calcium supplementation studies showed that increased dietary calcium in animals, infants, and children results in consistent decrease in the absorption of lead., Observed increase in serum calcium may be a physiological mechanism to decrease gastrointestinal absorption of lead.,
Naga Bhasma is indicated in Parinama Shula (duodenal ulcer), Shotha (edema), Kamala (jaundice), Vatarakta (gout), Grdhrasi (sciatica), Kustha (skin diseases), Apasmara (epilepsy), Undmada (hysteria), and Prameha (diabetes). All these diseases require long-term drug administration. Few studies on the safety of Naga Bhasma have been found in a review of previous research works however, in those works, Naga Bhasma was prepared by a minimum of 30 incineration cycles. Effect of Naga Bhasma prepared in seven incineration cycles on hematological and biochemical parameters was not studied. Therefore attempt was made to fulfill this gap. Another aim was also to elucidate the difference in safety profile with reference to change in media used for Bhasma preparation. As the work was conducted as per standard guidelines, therefore finding of this work may serve as a torchbearer in further research on similar subject.
| Conclusion|| |
The findings of hematological parameters presented major changes in MCV, MCH, and MCHC by NBH-treated group, whereas in biochemical parameters, both test drugs significantly affected BSL, SGOT, and serum ALP activity. Thus, Naga Bhasma prepared in seven incineration cycles by Parada media was found to be better than Vasa Swarasa media. However, both test drugs were not found completely safe, especially at higher dosage where changes related to renal and hepatic dysfunctions were found in hematological and biochemical analysis. Therefore, Naga Bhasma prepared in less number of incineration cycles is not therapeutically recommendable. Similar research works need to be conducted to validate findings of this research work.
Financial support and sponsorship
This work was supported by the IPGT and RA, Gujarat Ayurved University, Jamnagar, Gujarat, India.
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9], [Table 10], [Table 11], [Table 12], [Table 13], [Table 14], [Table 15], [Table 16], [Table 17], [Table 18]