Abstract
Purpose
This is the first comparative report that demonstrates the comparison of the anti-hyperglycemic activity of camel milk, buffalo milk and synthetic drugs in induced diabetic rabbits.
Method
Five groups (n = 8) of rabbits containing placebo (G1) and hyperglycemic groups (Alloxan® administered intravenously) including control diabetic (G2), camel milk treated @40 ml/kg (G3), buffalo milk treated @40 ml/kg (G4) and glibenclamide (Glicon®) @10 mg/kg (G5) orally for 60 days. Collection of blood was done for hematology and biochemical analysis. Renal and hepatic tissue sections were processed by routine paraffin technique for diabetes-induced histopathological changes and anti-diabetic activity of camel and buffalo milk.
Results
Diabetes deleteriously (P ≤ 0.05) affects all studied parameters. A significant (P ≤ 0.05) recovery was seen in diabetogenic hematological (RBC, MCV, Hb, MCH) and serological parameters (AST, ALT, creatinine, BUN, TPs, and TOS) with camel milk treatment. Camel milk and glibenclamide decreased blood glucose level more significantly (P < 0.01) than the buffalo milk but more significant renal recovery was seen by renal function. Microscopic observations demonstrated that camel milk and glibenclamide recovered the altered histology of the liver and kidneys towards normal.
Conclusion
The results indicate that camel milk has a potential therapeutic effect in the treatment of hyperglycemia and plays a significant role in its management as well as reduces the risk of diabetes-related complications as compared to buffalo milk.
Keywords: Diabetes, Alloxan, Camel milk, Buffalo milk, Glibenclamide, Creatinine
Introduction
Diabetes mellitus is a metabolic syndrome characterized by abnormally high blood glucose level, hyperglycemia, which developed due to faulty or low level of insulin with or without abnormal resistance to insulin. It is diagnosed by hyperglycemia in the fasting and postprandial state. Hyperlipidemia, protein wasting and ketosis are the major complication of severe and chronic dibetes. The worldwide incidence of diabetes mellitus appears to be increased and posing a major threat to global health. Poor glycemic control interrupts normal development and growth of children led to major complication in active life. Therefore, diagnosis, prevention and early treatment of hyperglycemia are important [1].
Traditionally, mammalian milk is a rich source of nutrients and considered a valuable component of a complete diet. Bovine milk contains many bioactive components that boost the physiological processes in the body. Furthermore, demand of food containing natural antioxidants is increased globally. The major antioxidants present in milk are grouped into lipid-soluble (retinol, carotenoids, and α-tocopherol) and water-soluble components (ascorbic acid) [2, 3]. Similarly, camel milk have imperative role in human nutrients especially habitants of arid and hot regions. The camel milk have high concentration of essential nutrients as compared to bovine milk [4]. Recently, many reports demonstrated its high therapeutic potential in non-communicable/metabolic diseases [5]. In the management of type 1 diabetes, the use of camel milk improved glycemic control of patients [6–8].
The anti-hyperglycemic potential of synthetic drugs (especially metformin, glibenclamide, and repaglinide) in the management of diabetes has been investigated extensively. Despite oral medication and other managemental strategies, diabetic complications including coma, hypoglycemic control, hepato-renal disturbances and immune response impairments are the main challenges of this condition [9].
For this purpose, the undertaken study was design to analyze the anti-hyperglycemic potential of camel milk in comparison with buffalo milk in Alloxan® induced hyperglycemic rabbits. Additionally, the renal and hepato protective effects of camel and buffalo milk in diabetic rabbits were also investigated.
Materials and methods
Animals
Forty rabbits, weighing about 1500–1800 g, were used for the study. The experiment was carried out following the guidelines of the Directorate of Graduate Studies and Institutional Animal Ethical Committee.
The rabbits were given basal diet consisted of casein (15%), corn oil (10%), cellulose (5%), salt mixture (4%), vitamin mixture (1%) and starch (65%) [10]. The water was available for animals during 24 h. The animals were kept in the animal house of the Department of Clinical Medicine and Surgery, University of Agriculture- Faisalabad for acclimatization in the maintained environmental conditions.
Therapeutic agents
Fresh raw Camel Milk (CM) was obtained on daily basis from local supplier of Anmol Marecha Dodh® at the time of milking. The constitutional analysis was carried out by the Livestock & Dairy Development Department of Punjab, Pakistan. Similarly, Buffalo milk (BM) was collected in raw form in sterile bottles and its chemical analysis was performed by the National Institute of Food Science and Technology (NIFSAT). The chemical composition of both these milk is given in Table 1. Both therapeutic agents were collected in sterile glass bottles and shifted to the experimental laboratory for administration to the rabbits.
Table 1.
Milk analysis of camel and buffalo done by the Livestock & Dairy Development Department (L&DDD) and National Institute of Food Science and Technology (NIFSAT) in collaboration with the University of Agriculture, Faisalabad
| Chemical Content (s) | Medicinal Content (s) | Minerals Content (s) | ||||||
|---|---|---|---|---|---|---|---|---|
| Component | Concentration | Component | Concentration | Component | Concentration | |||
| CM | BM | CM | BM | CM | BM | |||
| Water % | 86–88 | 85–87 | Cholesterol (mg/100 g) | 4.8–5.5 | 3.5–4.1 | Zinc (mg/100 ml) | 0.51–0.56 | 1.2–1.4 |
| Total Solids % | 12–13 | 13.5–15 | Vitamin C (mg/l) | 32.3–33.6 | 12.4–13 | Iron (mg/100 ml) | 10–10.3 | 0.37–0.4 |
| Fat % | 3.6–3.9 | 3–5 | vitamin E (mg/l) | 0.5–0.6 | 0.1–0.15 | Sodium (mg/100 ml) | 0.3–0.55 | 41–42 |
| Lactose % | 3.3–3.8 | 3–3.5 | Lactoferrin (mg/ml) | 0.32–0.44 | 0.12–0.15 | Potassium (mg/100 ml) | 149–153 | 9–10 |
| Protein % | 3.2–3.5 | 3.5–4 | Immunoglobulin G (mg/ml) | 1.6–1.7 | 0.65–0.70 | Copper (mg/100 ml) | 140–144 | 0.19–0.2 |
| Ash % | 0.7–1 | 0.8–1 | Insulin (IU/ml) | 50–52 | 16–17 | Calcium (mg/100 ml) | 110–114 | 107–109 |
CM Camel Milk, BM Buffalo milk
Diabetes induction
Diabetes was induced by a single intravenous administration of Alloxan® (Applichem, USA) and the dose rate of 160 mg/kg body was used in rabbits. The calculated dose of Alloxan® was mixed in normal saline and administrated to the experimental animals within 2 min. After 3 days of injection, the fasting blood glucose level was measured from ear vein puncture and animals having glucose level > 200 mg/dL were taken as diabetics and used for this study. The blood glucose level was measured using a FreeStyle® Optimum Neo Meter by Abbott Labs, Ltd. The normal or placebo animals were injected only with normal saline.
Research design
The induced diabetic or hyperglycemic rabbits were divided into four groups with eight biological replicates in each group. To compare with the normal, 8 rabbits were assigned to the placebo groups. The treatment was given to the experimental animals for 60 days. The description of different groups and their status are given in Table 2. Live body weight and glucose levels were measured weekly for evaluation of their therapeutic efficiency.
Table 2.
Description of research design used in this study
| Group | Diabetic Status | Treatment |
|---|---|---|
| G1 (Placebo) | Non-Diabetic; Only injected with vehicle intravenously | Normal diet and water |
| G2 (Diabetic control) | Induced diabetic groups | Normal diet and water |
| G3 (Camel milk treated) | Alloxan® was injected intravenously at the dose rate of 160 mg/kg. | Normal diet and water + camel milk orally at the dose rate of 40 ml/kg/day for 60 days |
| G4 (Buffalo milk treated) | Normal diet and water + Buffalo milk orally and dose rate of 40 ml/kg/day was for 60 days was used | |
| G5 (Glibenclamide treated) | Normal diet and water + Glibenclamideorally and the dose rate of 10 mg/kg/day for 60 days was used |
Collection of blood for hematological analysis
Before slaughtering of animals, blood samples (5 ml) were taken from jugular vein and shifted to the EDTA containing vacutainer for further analysis. Hematological profile, including hemoglobin level (Hb), packed cell volume (PCV), red blood cell (RBC) count, was determined by Medionic Hematology Analyzer (Germany) according to the user manual.
Serological parameters
Blood samples were centrifuged at 3000 rpm for 10 min, poured into labeled eppendrophs and stored at −20 °C to maintain enzyme activity.
Serum total oxidative stress (TOS)
TOS was determined by the colorimetric method as explained by Erel [11] after slight modifications.
Liver and kidney function test
Serum levels including urea, creatinine, uric acid, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, were determined using commercial kits from Bioclin®, according to the manufacturer’s protocol.
Collection of tissue samples and microscopic analysis
Animals were humanely slaughtered under gaseous anesthesia. These animals were opened for the collection of hepatic and renal tissue collection. Samples of liver and kidneys were transferred to fixative (neutral buffered formalin) immediately after washing with normal saline. Paraffin embedding technique [12] was employed on these tissue samples to prepare 5 micron thick sections that were subjected to Hematoxylin and Eosin (H&E) staining procedure. Microscopic slides were examined at 100 and 400 X. Vacuolar and degenerative changes in liver and kidney tissues were observed.
Statistical analysis
The data obtained were analyzed by Analysis of variance (ANOVA) technique using SPSS version 22 statistical computer software at 5% probability. Tukey’s honest significant test was used as post-ANOVA inference to compare the different groups.
Results
Bodyweight and blood glucose level
Post diabetic induction, the bodyweight of all groups decreased significantly (P < 0.05) than that of placebo group. Group G3 which fed with CM @40 ml/kg has significantly (P < 0.05) showed inclined trend of weight gain as compared to other treated groups (Fig. 4). The group administered with glibenclamide (Glicon)® @10 mg/kg orally also showed significant (P < 0.05) results in comparison to diabetic rabbits.
Fig. 4.
The first graph represents the trend of blood glucose level measured weekly in different groups. Camel milk and glibenclamide treated groups showed the declining trend of glucose level toward normal at every week of experimental. The second graph showed the trend of body weight gain during the experiment. In the last 3 weeks, the camel milk treated group significantly gained bodyweight (Sign “*” in a week represents the different groups significantly at P < 0.05)
Fasting blood glucose level of all groups are presented in Fig. 4. After induction of diabetes, significant (P < 0.05) elevated blood glucose level was seen in all induced diabetic groups. Treatment of CM, BM, and glibenclamide (Glicon®) treatment reduced significantly (P < 0.05) blood glucose levels. A continuous decrease in blood glucose levels has been shown by rabbits treated with CM@40 ml/kg throughout the experiment period.
Hematologic parameters
Hematological indices results are shown in the Table 3. Hyperglycemia led to significant (P ≤ 0.05) reduced RBC count and its indices in diabetic control group G2 as compared to control group G1. Diabetic altered levels of Red Blood Cells (RBCs), Hemoglobin (Hb), Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) were significantly (P < 0.05) improved by different treatment. CM and BM (P < 0.05) recovered RBCs indices but the more significant effect was seen in CM treated group (G3).
Table 3.
Therapeutic potential of camel milk and buffalo milk on hematological parameters in Alloxan®induced diabetic rabbits
| Groups | RBCs (1012/L) | MCV (fl) | Hb (g/dl) | MCH (pg) |
|---|---|---|---|---|
| G1 (Placebo) | 6.928 ± 0.07b | 60.4 ± 2.07b | 12.52 ± 0.52a | 18.36 ± 0.79a |
| G2 (Diabetic control) | 4.12 ± 0.86c | 49.16 ± 13d | 6.18 ± 4.04d | 12.05 ± 5.94d |
| G3 (Camel milk treated) | 5.86 ± 0.23ab | 63.3 ± 2.92b | 10.22 ± 5.04b | 14.96 ± 7.18b |
| G4 (Buffalo milk treated) | 5.036 ± 0.04b | 57.78 ± 1.48c | 7.48 ± 0.14c | 13.18 ± 6.67b |
| G5 (Glibenclamide treated) | 5.11 ± 0.16b | 59.48 ± 1.92c | 7.08 ± 4.04c | 14.00 ± 6.94b |
Different superscripts with different means statistically are different in a column at P ≤ 0.05 (Superscript “a” with the means is different from means b, c, d; Superscript “b” with the means is different from means a, c, d; Superscript “c” with the means is different from means a, b, d; Superscript “d” with the means is different from means a, b, c)
G1: Normal Negative Control, G2 Diabetic Positive Control, G3: Camel milk@40 ml/kg; G4: Buffalo milk @40 ml/kg; G5: Glibenclamide @10 mg/kg
Liver and kidney function tests and total proteins
Levels of Aspartate Amino-Transferase (AST) and Aspartate Aminotransferase (ALT) were observed significantly (P < 0.05) high in all diabetic group G2 in comparison to G1 (32.88 ± 1.13 and 31.10 ± 5.24 ul−1, respectively). All treatments, CM, BM and glibenclamide, showed significant (P < 0.05) antidiabetic effect but among these, camel milk gave more hepatic protection (Table 4). In kidney function tests like urea, uric acid and creatinine were significantly (P < 0.05) increased in diabetic induced group G2 (28.58 ± 1.92, 4.08 ± 0.33, 1.16 ± 0.27 mgdl−1, respectively) as compared to placebo groups (G1). Both CM and BM treatment significantly (P < 0.05) reduced these diabetic induced values of kidney function tests at the same significant level. In comparison, both CM and BM showed almost equal significant effect as the Glibenclamide treated (Table 4). Increased level of total protein in control diabetic group G2 was restored significantly (P < 0.05) towards normal values of group G1 in camel and buffalo milk treated group G3 and G4 at the same significant level. A similar anti-diabetic effect of CM and BM was observed in diabetic induced that caused decrease in albumin level (Table 4).
Table 4.
Therapeutic potential of camel milk and buffalo milk on liver and kidney function in Alloxan®induced diabetic rabbits
| Groups | ALT (ul−1) | AST (ul−1) | Total Protein (gdl−1) | Albumin (gdl−1) | Urea (mgdl−1) | Creatinine (mgdl−1) | Uric Acid (mgdl−1) |
|---|---|---|---|---|---|---|---|
| G1 (Placebo) | 7.38 ± 0.56b | 10.86 ± 2.49b | 7.74 ± 1.24b | 3.50 ± 0.55b | 28.58 ± 1.92b | 1.16 ± 0.27b | 4.08 ± 0.33b |
| G2 (Diabetic control) | 32.88 ± 1.13a | 31.10 ± 5.24a | 13.53 ± 0.72a | 2.58 ± 0.32a | 47.64 ± 5.28a | 2.11 ± 0.25a | 7.43 ± 0.87a |
| G3 (Camel milk treated) | 13.32 ± 1.53c | 14.48 ± 1.06c | 10.12 ± 0.74c | 4.00 ± 0.27c | 28.93 ± 1.98c | 1.59 ± 0.37b | 4.71 ± 0.12b |
| G4 (Buffalo milk treated) | 19.76 ± 1.14c | 16.55 ± 1.34c | 10.92 ± 0.79c | 3.05 ± 0.15c | 29.10 ± 1.99c | 1.59 ± 0.37b | 6.64 ± 0.44a |
| G5 (Glibenclamide treated) | 18.13 ± 0.95d | 12.02 ± 1.72b | 9.58 ± 0.54c | 3.85 ± 0.26b | 32.5 ± 2.18b | 1.15 ± 0.2c | 4.31 ± 0.21b |
Different superscripts with different means statistically are different in a column at P ≤ 0.05 (Superscript “a” with the means is different from means b, c, d; Superscript “b” with the means is different from means a, c, d; Superscript “c” with the means is different from means a, b, d; Superscript “d” with the means is different from means a, b, c)
G1: Normal Negative Control, G2 Diabetic Positive Control, G3: Camel milk@40 ml/kg; G4: Buffalo milk @40 ml/kg; G5: Glibenclamide @10 mg/kg
Total oxidative stress
Diabetes induction caused increased total oxidative stress as exhibited In Fig. 3. Treatment with CM and BM @40 ml/kg reduced TOS level significant (P ≤ 0.05) as compared to diabetic groups.
Fig. 3.
Comparison of the therapeutic effect of camel milk vs buffalo milk on Total Oxidative Stress in Alloxan®induced diabetic rabbits. Means sharing different superscript are statistically are different in a column at P ≤ 0.05 (Means having superscript “a” is different from means b, c, d; Means having superscript “b” is different from means a, c, d; Means having superscript “c” is different from means a, b, d; Means having superscript “d” is different from means a, b, c). G1: Normal Negative Control, G2 Diabetic Positive Control, G3: Camel milk@40 ml/kg; G4: Buffalo milk @40 ml/kg; G5: Glibenclamide @10 mg/kg
Histology
Histological examination of the diabetic group (G2) showed bi-nucleated hepatic cells, infiltration of inflammatory cells, cellular disruption, and derangement of hepatocytes around the central vein (Fig. 1b). The liver section of CM @ 40 ml/kg treated group (Fig. 1d) showed improvement in the arrangement of hepatic cells and decrease in inflammatory cells infiltration as compared to BM @ 40 ml/kg PO (Fig. 1c). The micrograph of glibenclamide (Glicon®) @10 mg/kg treated group showed normal restoration of hepatic cellular structure with a low load of inflammatory and binucleated cells. The results revealed that standard drug is more significant in comparison to camel milk and camel milk and showed significant results as compared to buffalo milk.
Fig. 1.
Photographs of hepatic tissue: Normal hepatic tissue (a) represents the normal hepatic cords (h) structure around central veins (cv) and portal triad (pt) while in diabetic rabbits (b) the structure of hepatic cells (h) is necrosed significantly (P < 0.05) and binucleated. The arrangement of his disturbed moreover lymphocytic infiltration (lf) is also seen. Treatment of buffalo milk (c) and camel milk (d) showed significant (P < 0.05) hepatic protective effect but more pronounced (P < 0.01) effect seen in camel milk treatment restoring normal hepatic structure around cv with no lf. (H&E, 100X)
Histological findings of normal renal tissues revealed the normal cellular structure of kidney corpuscles proximal and distal convoluted tubules (Fig. 2a). Sections of the control diabetic group (b) displayed tubular damage, shrinkage of the glomerulus, urinary space and presence of inflammatory cells (Fig. 2a). Histological examination of CM @40 ml/kg PO treated group (Fig. 2d) showed less degeneration of tubules and structure of kidney corpuscles closer to the normal structure as compared to the BM@40 ml/kg PO treated group (Fig. 2c). The kidney section of glibenclamide (Glicon®) @10 mg/kg treated group showed more improvement in glomerular and tubular degeneration. These results revealed that camel milk showed nephroprotective properties and improved the damaged kidney structure (Figs. 3 and 4).
Fig. 2.
Photographs of renal tissue: Normal renal tissue (a) represents the normal tubular structure of proximal (pct) and distal convoluted tubules (dct), glomerulus (g) and urinary space (us). Diabetes (b) caused severe (P < 0.05) tubular degeneration (td) shrinkage of g and us with lymphocytic infiltration (lf). Buffalo milk (c) did not show any protective effect on the renal tissue, however, camel milk treatment (d) exhibited (P < 0.05) restoring of normal tubular structure and glomerulus along with the urinary space. (H&E, 100X)
Discussion
Diabetes, metabolic syndrome, results in hyperglycemia due to faulty or deficient insulin secretion. It hampers the normal metabolism of the body, rises the free radical load that subsequently can cause oxidative stress and tissue damage in diabetic patients [13]. This experiment was designed to evaluate and compare the anti-diabetic therapeutic efficacy of camel (CM) and buffalo milk (BM) in induced diabetic rabbits. For the induction of diabetes in experimental animals, Alloxan is extensively used because of its selective cytotoxic activity towards beta cells of the pancreas and also diminishes antioxidant capacity [14]. Alterations in the RBCs and Hb, level in hyperglycemic conditions suggest the occurrence of anemia. Diabetic induced anemic effects on erythrocytic indices was in accord with Erukainure et al. [15], Usman et al. [16] and Qureshi et al. [17] findings. Anemic conditions in diabetes has been reported due to decline in erythropoietin synthesis which leads to kidney failure, elevated non-enzymatic glycosylation of RBCs membrane proteins and other hematological indices (Hb, MCV, MCH). Anti-hyperglycemic effect of CM significantly (P < 0.05) recovered diabetic altered RBCs and Hb towards normal as compared to BM. These recovered parameters of CM fed rabbits suggests its potency in the management of the ailment. This could be related to its high concentration of minerals and vitamins that activate the anti-oxidant system. Thus, preventing oxidation stress on RBCs and Hb that caused hemolysis [18]. Results are in line with the finding of of Agrawal et al. [7] and Sing [19] for the hypoglycemic effect of CM in diabetes. A distinctive characteristic of CM is its high concentration (52 units/l) of insulin as compared to other ruminant’s milk and its quality to impede coagulum formation in the stomach or the acidic media, thereby preventing degradation of insulin in the stomach. Further investigations [6, 20] found that amino acid sequences CM proteins are rich in half cysteine that possess superficial similarity with the family of insulin peptides [21]. This unique property of CM can be attributed to anti-hyperglycemic potential and protector that stimulate the absorption of intact insulin molecules in small intestine. All these above mentioned factors may combine and contribute towards hypoglycemic potential of CM in current study.
The beneficial health effects of CM were observed in the liver and kidney functions in diabetic patients. The elevated liver enzyme, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), are common seen in patients with diabetes mellitus. These enzymes reflect the picture of enzyme concentration of intracellular hepatic that might have leaked and come into the circulation due to diabetic induced hepatic injury. The findings in the present study are consistent with Hamad et al. [22] and Ali et al. [23]. Camel milk treatment significantly protected the hepatic parameters in diabetic rabbits as compared to BM. Diabetogenic nephropathy is microvascular in nature and is taken as an important diabetes complication. The presence of urea and proteins are predictor of poor renal outcome. CM has a potential role in maintaining micro-albuminuria levels in type-I diabetic. Level of microalbuminuria has been reduced from 119.48 ± 1.68 mg/dL to 22.52 ± 2.68 mg/dL in type-I diabetic patients by addition of CM treatment to the usual diet for 6 months. Kidney function indicators (uric acid, urea and creatinine) were moved significantly towards normal level in CM fed diabetic rabbits [6]. Creatinine levels increased in diabetes (type I and II) due to renal arterial disorder and/or cardiac failure. Failure of excretion of toxic metabolic waste products of proteins may also cause renal disorders like in diabetes [15].
In the current study, reduced level of liver function enzymes and kidney function indicators in the CM treated diabetic group showed its therapeutic efficiency of against hepato- and renal toxicity. Elevated levels of these enzymes have been reported in diabetic rabbits. This enhanced level might be because of leakage from the cytoplasm into circulation after the oxidation and injury of cellular membrane and damage. Improvement of urea and creatinine level in rabbits administered with CM and BM reflected improved kidney function and decreased destructive effect of Alloxan® Increased creatinine level associated with decreased level of Hb leading to anemia. This creatinine decline in the hyperglycemic rabbits was inversely related to the Hb level, thus correlating with the antianemic efficacy of the camel milk [15]. Buffalo milk showed a significant protective effect on the kidneys function test. This nephroprotective effect was probably due to the biologically active peptides present in the whey proteins [14].
The present study showed the oxidative stress is directly related to the diabetic manifestations. Increased oxidative stress in hyperglycemic patients decrease the non-enzymatic antioxidants levels such as glutathione, Vita E and Vit C that subsequently hampers metabolic pathways and caused diabetic complications. In this trial total oxidative stress reduced in rabbits treated with CM, BM, and glibenclamide. This effect of CM and BM may be linked with protective efficacy against lipid peroxidation, thereby contributing towards the protection against oxidative stress and damage in Alloxan® induced diabetes in animals. The administration of sulfonylurea oral class of hypoglycemic drugs exhibited a significant reduction in total oxidative stress and improved the decrased activities of antioxidant enzymes.
Insulin, also acts as an anti-inflammatory agent, along with the higher level of zinc in camel milk may have a role in the stimulation of the anti-oxidant system in the body through glutathione. Glibenclamide also provides additional defense against anti-oxidant, thereby protects the pancreas from stress induced damage during diabetes complications as stated by Chukwunonso et al. [24]. Another study by EL-Said et al. [25] evaluated the therapeutic effect of CM on oxidation induced stress in hyperglycemic rabbits.
Those hyperglycemic rabbits treated with CM showed significantly (p < 0.05) improve these levels of malondialdehyde (5.6 ± 0.3 nmol/mL), catalase (377.5 ± 4.2 U/L,), and glutathione (10.1 ± 0.7 mg/dL) than that of untreated diabetic rabbits malondialdehyde (8.7 ± 0.2 nmol/L), catalase (204.7 ± 17.9 U/L), and glutathione (8.6 ± 0.6 mg/dL).The protective effects of CM might be due to its ant-ioxidant and chelating effects on toxicants. CM contains higher concentration of vitamins (A, B2, E, and C) and mineral content (e.g., Na, K, Cu, Mg, and Zn).
The vitamins and minerals act as antioxidants and are useful in prevention against cellular injury caused by toxic agents’ liks Alloxan® and streptozotocin and hence decrease the load free radicals. Additionally, camels prefer grazing on natural vegetation containing in particular, salty plants, desert bushes and herbs. This kind of diet may also have pivotal role in providing some of the phytochemicals excreted in CM that give additional therapeutic effect to diabetic patients [26].
Alloxan® increased the load of free radical due to which hepato renal tissues become more susceptible to oxidative injury and severely affect the normal cellular structure in the hyperglycemic rabbits as stated in literature [17, 25].
The cellular degenerations of renal and hepatic tissues, observed in the present study, were following the Ragavan and Krishnakumari [27] and Ateeq et al. [18].
Camel milk and glibenclamide significantly recovered the normal cellular architecture of tissues, the more protection was observed in glibenclamide treated animals. The presence of antioxidant vitamins (A, B, C, and E), insulin and zinc in CM [28] might be held responsible for the recovery of diabetogenic alterations in hepatic and renal structure towards normal.
Conclusion
It is conceivable from the current data that the hematobiochemical findings of the present study are confirmed with histopathological alterations observed in the rabbits treated with either camel milk, buffalo milk or glibenclamide(Glicon)®. It is well demonstrated that camel milk has higher therapeutic efficacy as compared to buffalo milk against hematobiochemical changes associated with induced diabetes in rabbits. However, further studies are warranted to identify the active components present in camel milk and to find out their exact mode of action.
Authors contribution
F Deeba and AS Qureshi gave the idea of this research M Kamran performed the research trail. F Deeba, AS Qureshi and N Faisal supervised the lab work. A Farooq and H Muzzafar helped in analyzing of blood parameters. M Usman and M Kamran collected the data and applied statistics. The manuscript was finalized by AS Qureshi and M Usman.
Compliance with ethical standards
Conflict of interest
This article is submitted with the consent of all co-authors and authors have no conflict of interest to declare.
Ethical approval
All procedures performed in this study are following the research ethical standards of the University of Agriculture, Faisalabad, Pakistan.
Footnotes
Highlights
This is the first comparative report that demonstrates the comparison of the anti-hyperglycemic activity of camel milk, buffalo milk and synthetic drugs in induced diabetic rabbits. This article also highlights the anti-hyperglycemic properties of camel milk that might be used for management glycemic control and its associated hematological, renal and hepatic complications.
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
References
- 1.Dahlquist GG. Primary and secondary prevention strategies of pre-type 1 diabetes. Diabetes Care [Internet]. 1999 [cited 2019 Dec 26];22(4):6. Available from: https://search.proquest.com/openview/5b755f5ed92cbce2b699554896c17b3c/1?cbl=47715&pq-origsite=gscholar. [PubMed]
- 2.Khan IT, Bule M, Ullah R, Nadeem M, Asif S, Niaz K. The antioxidant components of milk and their role in processing, ripening, and storage: functional food. Vet World. 2019;12(1):12–33. doi: 10.14202/vetworld.2019.12-33. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3.Haug A, Høstmark AT, Harstad OM. Bovine milk in human nutrition - a review. Lipids Health Dis. 2007;6:25. doi: 10.1186/1476-511X-6-25. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Omer RH, Eltinay AH. Changes in chemical composition of camel’s raw milk during storage. Pak J Nutr. 2009;8(5):607–610. doi: 10.3923/pjn.2009.607.610. [DOI] [Google Scholar]
- 5.Yateem A, Balba MT, Al-Surrayai T, Al-Mutairi B, Al-Daher R. Isolation of lactic acid Bacteria with probiotic potential from camel milk. Int J Dairy Sci. 2008;3(4):194–199. doi: 10.3923/ijds.2008.194.199. [DOI] [Google Scholar]
- 6.Agrawal RP, Jain S, Shah S, Chopra A, Agarwal V. Effect of camel milk on glycemic control and insulin requirement in patients with type 1 diabetes: 2-years randomized controlled trial. Eur J Clin Nutr. 2011;65(9):1048–1052. doi: 10.1038/ejcn.2011.98. [DOI] [PubMed] [Google Scholar]
- 7.Agrawal RP, Kochar DK, Sahani MS, Tuteja FC, Ghorui SK. Hypoglycemic activity of camel milk in streptozotocin induced diabetic rats. International Journal of Diabetes in Developing Countries. 2004;24:47–49. [Google Scholar]
- 8.Al haj OA, Al Kanhal HA. Compositional, technological and nutritional aspects of dromedary camel milk. Int Dairy J. 2010;20:811–821. doi: 10.1016/j.idairyj.2010.04.003. [DOI] [Google Scholar]
- 9.Chukwunonso Obi B, Chinwuba Okoye T, Okpashi VE, Nonye Igwe C, Olisah Alumanah E. Comparative study of the antioxidant effects of metformin, glibenclamide, and repaglinide in alloxan-induced diabetic rats. J Diabetes Res. 2016;2016:1635361. doi: 10.1155/2016/1635361. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Peterson DT, Greene WC, Reaven GM. Effect of experimental diabetes mellitus on kidney ribosomal protein synthesis. Diabetes. 1971;20(10):649–654. doi: 10.2337/diab.20.10.649. [DOI] [PubMed] [Google Scholar]
- 11.Erel O. A novel automated method to measure total antioxidant response against potent free radical reactions. Clin Biochem. 2004;37(2):112–119. doi: 10.1016/j.clinbiochem.2003.10.014. [DOI] [PubMed] [Google Scholar]
- 12.Suvarna S, Layton C, Bancroft JD. Bancroft’s theory and practice of histological techniques. Churchill Livingstone Elsevier. 2013;345–350
- 13.Alimohammadi S, Hobbenaghi R, Javanbakht J, Kheradmand D, Mortezaee R, Tavakoli M, et al. Protective and antidiabetic effects of extract from Nigella sativa on blood glucose concentrations against streptozotocin (STZ)-induced diabetic in rats: an experimental study with histopathological evaluation. Diagn Pathol [Internet]. 2013 Dec 15 [cited 2019 Dec 27];8(1):137. Available from: https://diagnosticpathology.biomedcentral.com/articles/10.1186/1746-1596-8-137. [DOI] [PMC free article] [PubMed] [Retracted]
- 14.Hassan Bisar G, Youssef M, El Saadany K, El Kholy WM, Kheadr E. Effect of lentil and Buffalo whey protein Hydrolysates on histopathology of liver and kidney in diabetic rats. J Cytol Histol. 2017;08(05):489. doi: 10.4172/2157-7099.1000489. [DOI] [Google Scholar]
- 15.Erukainure OL, Ebuehi OAT, Adeboyejo FO, Aliyu M, Elemo GN. Hematological and biochemical changes in diabetic rats fed with fiber-enriched cake. J Acute Med. 2013;3(2):39–44. doi: 10.1016/j.jacme.2013.03.001. [DOI] [Google Scholar]
- 16.Usman M, Ali MZ, Qureshi AS, Ateeq MK, Nisa FU. Short term effect of dose dependent camel milk in alloxan induced diabetes in female albino rats. J Anim Plant Sci. 2018;28(5):1292–1300. [Google Scholar]
- 17.Qureshi AS, Ghaffor J, Usman M, Ehsan N, Umar Z, Sarfraz A. Effect of ethanolic preparations of cinnamon (Cinnamomum zeylanicum) extract on hematologic and histometric parameters of selected organs in Alloxan® induced diabetic female albino rats. J Diabetes Metab Disord. 2019. 10.1007/s40200-019-00457-4. [DOI] [PMC free article] [PubMed]
- 18.Ateeq MK, Qureshi AS, Usman M, Shahid RU, Khamas WA. Effect of orally administered camel milk in Alloxan® induced albino rats: long term study on maternal uterus and neonates selected organs. Pak Vet J. 2019;39(1):81–85. doi: 10.29261/pakvetj/2018.114. [DOI] [Google Scholar]
- 19.Singh R. Influence of intensive diabetes treatment on quality-of-life outcomes in the diabetes control and complications trial. Diabetes Care. 1996;19(3):195–203. doi: 10.2337/diacare.19.3.195. [DOI] [PubMed] [Google Scholar]
- 20.Abdalla KO. An overview of the therapeutic effects of camel milk in the treatment of type 1 diabetes mellitus. J Biomol Res Ther. 2014;03(03):118–124. [Google Scholar]
- 21.Beg OU, von Bahr-Lindström H, Zaidi ZH, Jörnvall H. Characterization of a camel milk protein rich in proline identifies a new β-casein fragment. Regul Pept. 1986;15(1):55–61. doi: 10.1016/0167-0115(86)90075-3. [DOI] [PubMed] [Google Scholar]
- 22.Hamad EM, Abdel-Rahi EA, Romeih EA. Beneficial effect of camel milk on liver and kidneys function in diabetic Sprague-dawley rats. Int J Dairy Sci. 2011;6(3):190–197. doi: 10.3923/ijds.2011.190.197. [DOI] [Google Scholar]
- 23.Ali MZ, Qureshi AS, Usman M, Kausar R, Ateeq MK. Comparative effect of camel milk and black seed oil in induced diabetic female albino rats. Pak Vet J. 2017;37(3):293–298. [Google Scholar]
- 24.Chukwunonso Obi B, Chinwuba Okoye T, Okpashi VE, Nonye Igwe C, Olisah AE. Comparative study of the antioxidant effects of metformin, glibenclamide, and repaglinide in alloxan-induced diabetic rats. J Diabetes Res. 2016;2016:1–5. doi: 10.1155/2016/1635361. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25.El-Said EE, EL-Sayed GR, Tantawy E. Effect of camel milk on oxidative stresses in experimentally induced diabetic rabbits. Vet Res Forum. 2010;1(1):30–43. [Google Scholar]
- 26.Shori AB. Camel milk as a potential therapy for controlling diabetes and its complications: a review of in vivo studies. J Food Drug Anal. 2015;23(4):609–618. doi: 10.1016/j.jfda.2015.02.007. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27.Ragavan B, Krishnakumari S. Effect of T. arjuna stem bark extract on histopathology of liver, kidney and pancreas of alloxan-induced diabetic rats. Afr J Biomed Res. 2009;9(3):189–197. [Google Scholar]
- 28.Mullaicharam A. World journal of pharmaceutical sciences a review on medicinal properties of camel milk A . R . Mullaicharam. World J Pharm Sci. 2014;2(3):237–242. [Google Scholar]