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Research Article - (2023) Volume 15, Issue 4

COMPLEXATION, CHARACTERIZATION, AND TOXICOLOGY STUDIES OF THE CRUDE EXTRACT OF ANACARDIUM OCCIDENTALE LEAVES AND ITS COPPER (II) COMPLEX

Akinola Dolapo Esther1, Oladipo Adelaide Mary1, Ajao Folasade Omobolanle2* and Iyedupe Marcus Olaoye2
 
1Department of Chemistry, Faculty of Pure and Applied Sciences, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Oyo State, Nigeria
2Physiology Department, Faculty of Basic Medical Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Oyo State, Nigeria
 
*Correspondence: Ajao Folasade Omobolanle, Physiology Department, Faculty of Basic Medical Science, Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Oyo State, Nigeria, Tel: +2348034659049, Email:

Received: 01-Jun-2023, Manuscript No. ijddr-23-13798; Editor assigned: 05-Apr-2023, Pre QC No. ijddr-23-13798 (PQ); Reviewed: 19-Apr-2023, QC No. ijddr-23-13798; Revised: 23-Jun-2023, Manuscript No. ijddr-23-13798 (R; Published: 30-Jun-2023

Abstract

Aim and objective: The search for effective therapy for diabetes mellitus is still unattainable. This study investigated anti-diabetic effect of Anacardium Occidentale Ethanolic leaves extract and Ethanolic leaves extract complex copper (II) in alloxan-induced diabetic rats.

Method: Fifty six matured male Wistar rats (200-250g) were used. Alloxan monohydrate (65 mg/kgb.wt) was singly injected intraperitoneally to induced diabetes. The animals were randomly alienated into 7groups, 8rats/group. Group I: normal control; Group II: diabetic; Group III & IV: diabetic rats + 400 mg/kgb.wt and 600 mg/kgb.wt A. Occidentale Ethanolic crude leaves extract; Group V and VI: diabetic rats + 400 mg/ kgb.wt and 600 mg/kgb.wt Ethanolic leaves extract complex copper (II); Group VII: diabetic rats + 500 mg/kgb.wt metformin for 15 days. Body weight and fasting blood glucose (FBG) were recorded at day 0, 5th, 10th, and 15th of the experiment. The animals were sacrificed at the end of the experiment and fasting blood samples were collected for biochemical parameters assay.

Results: Malondialdehyde (MDA), FBG, lipid profile, hepatic and kidneys function biomarkers levels were significantly (p<0.05) increased in the diabetic rats while the body weight and antioxidant activities were significantly (p<0.05) reduced. Administrations of A. Occidentale Ethanolic leaves extract, Ethanolic leaves extract complex copper (II) and metformin significantly (p<0.05) diminished the FBG, MDA, lipid profile, hepatic and kidney function biomarkers levels and elevated the body weight and antioxidant activities.

Conclusion: The Ethanolic leaves extract complex copper (II) possesses more anti-diabetic efficacies. This could be explored as effective complementary therapy for diabetes mellitus.

Keywords

A. Occidentale leaves; Diabetes mellitus; Dyslipidemia; Leaves extract complex copper (II); Oxidative stress

INTRODUCTION

Diabetes mellitus is considered as one of the three noncommunicable metabolic diseases with an alarming increase in prevalence globally. According to the International Diabetes Federation (IDF) estimation, the number of diabetes patient is projected to rise from 425 million to 629 million by 2045.

Diabetes mellitus is endocrine metabolic disorder characterized by chronic hyperglycemia resulting from deficient insulin secretion from beta-cells (β-cell) of pancreas (type-1) or irresponsiveness of target cells to insulin action (type-2) or both, thereby affecting metabolism of carbohydrates, proteins and fats. Diabetic patients suffered from type -2 diabetes mellitus is about 95 percent. Clinical symptoms and diagnosis of diabetes mellitus include body weight loss, fatique, polyphagia, ployuria, and polydipsia manifested with hyperglycemia.

Chronic hyperglycemia in diabetic state increases the risk and progression of micro and macro-vascular diabetic complications such as nephropathy, neuropathy, retinopathy, atherosclerosis, cardiovascular disease and stroke, reducing life quality and expectancy.

Hyperglycemia-induced hyperlipidemia and oxidative stress in terms of abnormal lipid profile and generation of excessive free radicals with suppression in endogenous antioxidants defense system play a central role in the pathogenesis of diabetes mellitus complications [1].

Generally, the available anti-diabetic drugs have many drawbacks owing to their numerous adverse toxicities, leading to an increasing search for a safe and non-toxic therapy for diabetes. The use of natural product and their purified compounds from medicinal plants has now gain popularity as promising and novel therapy for diabetes mellitus as it is easily accessible and could be safe from toxic effect.

Anacardium Occidentale Linn (family Anacardiaceae) alternatively known as cashew tree originated from Brazil. Phytochemical screening analysis of various part of A. Occidentale revealed the presence of numerous bioactive compounds (tannins, flavonoids, phenols, saponins, alkaloids, and anacardic). A. Occidentale leaf, nut, root and stem bark are medicinally used for management of different maladies such as diabetes, infection, diarrhea, hemorrhage, and as well as antimicrobial activity. The nut has been reported to attenuate dyslipidemia in rats. Also, anti-hyperglycemic and anti-oxidative of A. Occidentale methanolic leaves extract and Ethanolic stem bark extract has been reported from in vivo studies. Beside the utilization of plant extracts as therapeutic agent for diabetes, several studies reported the role of essential metals in glucose metabolism regulation and their deficiency associated with the risk of diabetes.

Copper, the third most abundant essential metal in humans involved in oxidative stress processes regulation. Evidence from literature showed that deficiency of this metal leading to many diabetes complications. However, research on complexation of A. Occidentale part with medicinal metal oxide for diabetes and its related complications progression therapy has never been reported. Therefore, this study investigates the antidiabetic activity of A. Occidentale Ethanolic leaves extract and extract complexed with copper (II) in alloxan-induced diabetic ra>ts model [2].

MATERIALS AND METHODS

Chemicals and drugs: Alloxan monohydrate, glucose solution, ketamine, Xylazine, Ethanol

Plant material: Anacardium Occidentale leaves were collected from the Agricultural Research Farm, Ladoke Akintola University, Ogbomoso, and Oyo State, Nigeria. The plant was identified, authenticated and given a voucher number LH0533 by Dr. A. T. J. Ogunkunle at Biology Department, Ladoke Akintola University of Technology.

Preparation of A. occidentale crude ethanolic leaves extract and ethanolic leaves extract complex copper (ii)

The leaves of A. occidentale were washed thoroughly and air-dried at room temperature. The leaves were grinded into a fine-powdered form using an electric blender and stored in air-tight container. 500 g of the fine powdered form was extracted in a Soxhlet apparatus using ethanol solvent (95%).The filtrate was then evaporated to dryness under reduced pressure in a rotary evaporator and the ethanolic extract obtained was kept at 4oC in an air-tight container until used.

The ethanolic leaves extract copper (II) complex was prepared by dissolving 0.5 g (2.75x10-3 mole) of copper (II) acetate in 42 cm3 of distilled water. The copper acetate solution was added drop wisely to a solution of 1 g A. occidentale crude extract in 50 cm3ethanol. The solution was rigorously stirred using a magnetic stirrer at 3500 rpm for 1hr. The precipitated complex was filtered and washed with water distilled water then dried in vacuum desiccators over calcium chloride (Cacl2). A pure green solid was obtained and weighed [3].

Experimental animals: Fifty six matured male Wistar albino rats weighing (200-250 g) were procured and housed at Department of Physiology Animal Research House, Ladoke Akintola University of Technology, Ogbomoso, Oyo state, Nigeria. The animals were kept in a well-clean ventilated plastic cage and allotted into seven groups, eight rats each. The animals were fed with a standard feed and water ad libitum under pathogen-free environment of temperature (25 ± 2oC), relative humidity (40 ± 5oC) and natural 12:12 light/dark cycle one week acclimatization prior the initiation of the experiment. The experimental procedures were conducted according to the National Guide for the Care and Use of Laboratory Animals and approved by Ladoke Akintola University of Technology Research Committee

Diabetes induction: The animals were fasted overnight (12hrs) before diabetes induction. After an overnight fasting, forty eight animals were injected intraperitoneally with a single dose (65 mg/kgb.wt) alloxan monohydrate dissolved in a normal saline (Nacl) to induced diabetes and allow to drink 5% glucose solution to prevent druginduced hypoglycemic death. After 72hrs of alloxan injection, fasting blood glucose levels of the animals were checked with glucometer (Accu-check) using tail vein blood dropped wisely on test stripe to authenticate diabetes induction. Animals with fasting blood glucose levels ≥ 250 mg/dl were chosen as diabetic rats and used for the study. The fasting blood sugar levels of the animals were recorded prior the injection of alloxan monohydrate [4].

Experimental design and treatment

The animals were randomly divided into seven groups, 8 rats each.

Group I: Normal control (non-diabetic)

Group II: Diabetic (untreated) rats.

Group III: Diabetic rats + 400 mg/kgb.wt A. occidentale ethanolic leaves extract

Group IV: Diabetic rats + 600 mg/kgb.wt A. occidentale ethanolic leaves extract

Group V: Diabetic rats + 400 mg/kgb.wt A. occidentale ethanolic leaves extract complex copper (II)

Group VI: Diabetic rats + 600 mg/kgb.wt A. occidentale ethanolic leaves extract complex copper (II)

Group VII: Diabetic rats + 500 mg/kgb.wt metformin.

The extracts and metformin were given orally with the aid of cannula for 15 consecutive days with feed and water ad libitum.

The body weight and fasting blood glucose levels of the animals were measured at day 0, 5th, 10th, and 15th of the experimental period. Fasting blood glucose levels of the animals were measured by glucose oxidase-peroxidase (GOD-POD) method using one-touch digital glucometer (Accu-chek) by dropping tail vein blood on glucose test stripes.

Biochemical assay: At the end of the 15 days treatment, the animals were subjected to overnight fasting and sacrificed by cervical dislocation after anesthetized with (40 mg/kgb. wt) and xylazine (15 mg/kgb.wt) injected intraperitoneally. Fasting blood samples were collected from the animals apex beat heart via cardiac punctured into heparinized tubes and centrifuged at 3500 rpm for 5 minutes. The clear supernatant plasma collected after centrifugation was used for biochemical parameters estimations.

Triglycerides (TG), total cholesterol (TC) and high density lipoprotein-cholesterol (HDL) levels were estimated based on enzymatic colorimetric method with a commercial Diagnostic Kit (Genzyme Diagnostics, MA, USA) according to the manufacturers’ protocol. Low-density lipoprotein-cholesterol (LDL) level was calculated using Friedewald et al. formula; LDL = TC – (HDL - TG/5) [5]. Oxidative stress marker malondialdehyde (MDA) level and activities of antioxidants superoxide dismutase (SOD), and catalase (CAT) were measured by enzyme linked Immunosorbent assay (ELISA) methods using specific Rat MDA, SOD and CAT commercial Elisa Kit (Elabscience, China) follow manufacturers’ instruction. Glutathione reductase (GSH) was measured based on the Gupta and Gupta method.

Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) hepatic enzymes biomarkers levels were measured spectrophotometrically using standard automated techniques based on the instruction of the manufacturer.

Urea and creatinine kidney function markers levels were determined using a commercial assay kit obtained from Siemens Health Care Diagnostics.

Statistical analysis: Statistical Package for Social Sciences (SPSS version, 21.0) was used to analyze the data. The data values were expressed as standard error of mean (mean ± SEM) and data statistical significant was evaluated using one-way analysis of variance (ANOVA) followed by Tukey’s post-hoctest. Data at P<0.05 were considered statistically significant [6].

RESULTS

Effect of A. occidentale ethanolic leaves extract and complex extract on body weight change in alloxan-induced diabetic rats

The body weight of untreated diabetic rats significantly (p<0.05) decrease throughout the experiment compared with the control group. However, administration of the A. occidentale ethanolic leaves extract and complex extract significantly increase the body weight compared with untreated groups (Tab. 1a.).

  PRE-TREATMENT             POST- TREATMENT  
GROUPS DAY 0 DAY 5 DAY 10 DAY 15
Normal (non-diabetic) 239.67 ± 51.98 245.33 ± 52.60 246.00 ± 10.44 251.00 ± 8.54
Diabetic (untreated) 239.33 ± 45.39 * 227.67 ± 42.71* 214.00 ± 41.61* 195.67 ± 42.92 *
Metformin  (500 mg/kg) 228.67 ± 21.73# 212.00 ± 20.30# 223 ± 19.00# 238.00 ± 17.06#
Crude extract (400 mg/kg) 223.67 ± 16.20# 229.00 ± 12.17# 234.00 ± 9.45# 247.00 ± 9.30#
Crude extract (600 mg/kg) 286.67 ± 14.57# 281 ± 12.77# 283.67 ± 12.90# 290.00 ± 13.45#
Complex extract (400 mg/kg) 216.00 ± 3.61# 221.67 ± 4.04# 226.33 ± 4.04# 232.67 ± 4.04#
Complex extract (600 mg/kg) 221.00 ± 3.61# 226.33 ± 4.51# 222.67 ± 4.51# 227.67 ± 4.16#      

Tab.1a. Effect of A. Occidentale Ethanolic leaves extract and complex extract on body weight change in alloxan-induced diabetic rats. Values are expressed as mean ± SEM (n=8). *significant at p<0.05 compared with control; #significant at p<0.05 compared with untreated diabetic group.

Effect of A. occidentale ethanolic leaves extract and complex extract on fasting blood glucose levels in alloxan-induced diabetic rats

Fasting blood glucose levels of untreated diabetic rats significantly (p<0.05) elevated compared with the control group. Supplementation of A. occidentale ethanolic leaves extract and complexed extract as well as metformin significantly reduced the fasting blood glucose level in comparison with the untreated diabetic group. Better hypoglycemia result was obtained in the diabetic group treated with high dose of the complex extract (600 mg/ kgb.wt) (Tab. 1b.).

  PRE-TREATMENT POST-TREATMENT
GROUPS DAY 0 DAY 5 DAY 10 DAY 15
Normal (non-diabetic) 100.33 ± 15.01 98.00 ± 14.53 101.00 ± 14.11 104.00 ± 14.01
Diabetic (untreated)  236.00 ± 10.58* 240.00 ± 10.54* 276.00  ± 15.28* 263.33 ± 6.51*
Metformin (500 mg/kg) 314.67 ± 11.68# 283.33 ± 21.73# 259.67 ± 44.80# 193.67 ± 12.12# 
Crude extract (400 mg/kg) 324.67 ± 8.50# 329.00 ± 9.54# 191.33 ± 23.84# 151.00 ± 23.50#
Crude extract (600 mg/kg) 344.67 ± 18.82# 312.33 ± 9.61# 192.67 ± 7.51# 144.00 ± 32.36#
Complex extract (400 mg/kg) 331.33 ± 40.70# 343.00 ± 69.59# 272.00 ± 59.86# 220.00 ± 34.39#
Complex extract (600 mg/kg) 312.67 ± 3.06# 281.00 ± 36.67# 184.00 ± 45.62# 138.00 ± 20.82#

Tab.1b. Effect of A. Occidentale Ethanolic leaves extract and complex extract on plasma fasting blood glucose in alloxan-induced diabetic rats. Values are expressed as mean ± SEM (n=8). *significant at p<0.05 compared with control; #significant at p<0.05 compared with untreated diabetic group.

Effect of A. occidentale ethanolic leaves extract and complex extract on lipid profile in alloxan-induced diabetic rats

The untreated diabetic rats showed significant (p<0.05) elevation in triglyceride (TG), total cholesterol (TC) and low-density lipoprotein (LDL) levels with significant decrease (p<0.05) in high-density lipoprotein (HDL) level. Administration of A. Occidentale Ethanolic leaves extract, complex extract and metformin to alloxan-induced diabetic rats markedly decline TC, TG, LDL levels along with a marked increase in HDL level compared to the untreated group (p<0.05). The high dose of the complex extract (600 mg/kgb.wt) exhibited more significant lowering effect on TC, TG, LDL levels to the extent of approaching control level and elevates HDL-c level reaching the level of control group (Fig.1.).

figure

Figure 1:Effect of A. occidentale ethanolic leaves extract and complex extract on lipid profiles in alloxan-induced diabetic rats. Values are expressed as mean ± SEM (n=8). *significant at p<0.05 compared with control; #significant at p<0.05 compared with untreated diabetic group.

Effect of A. occidentale ethanolic leaves extract and complex extract on oxidative stress marker and antioxidants enzymes activity and in alloxan-induced diabetic rats

Diabetic rats exhibited signification (p<0.05) reduction in the activities of enzymatic antioxidants superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) with significant (p<0.05) increase in oxidative stress marker malondialdehyde (MDA) level in comparison with the control group. The antioxidants enzymes SOD, CAT, and GSH activities were up-regulated along with downregulation in the MDA level after administration of crude extract of A. Occidentale, complexed extract and as well as metformin to the diabetic rats compared to untreated group. Low dose of the complex extract (400 mg/kgb. wt) offered best result in the up-regulation of antioxidant enzymes with value more than the control group and then other doses and metformin (Tab. 2.).


GROUPS
MARKERS Control Diabetic (untreated) Low Extract (400mg/kgb.wt) High Extract (600mg/kgb.wt) Low Complex (400mg/kgb.wt) High Complex (600mg/kgb.wt) Metformin (500mg/kgb.wt)
MDA (µmol/g) 35.69 ±1.31 68.81  ± 3.75* 39.07 ± 5.01# 38.67 ± 2.60# 36.26 ± 2.93# 35.75± 1.10# 34.08 ± 1.72#
GSH  (µmol/g) 0.60 ± 0.17 0.49  ± 0.19* 0.70 ±0.08# 0.60 ±0.004# 0.74 ±0.05# 0.57±0.01# 0.59 ± 0.03#
SOD (µmol/g) 1.29 ± 0.03 0.59  ± 0.08* 1.30 ±0.04# 1.26 ± 0.008# 1.34 ±0.08# 1.24± 0.01# 1.25 ± 0.01#
CAT (µmol/g) 7.29 ± 0.23 3.14  ± 0.34* 7.35 ±0.40# 7.12 ±0.28# 7.46 ±0.27# 7.28±0.19# 7.13 ±0.16#

Tab.2. Effect of A. Occidentale Ethanolic leaves extract and complex extract on oxidative stress marker and antioxidants enzymes activity and in alloxaninduced diabetic rats. Values are expressed as mean ± SEM (n=8). *significant at p<0.05 compared with control; #significant at p<0.05 compared with untreated diabetic group.

Effect of A. occidentale ethanolic leaves extract and complex extract on liver and kidney function markers in alloxaninduced diabetic rats

Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, (ALP), albumin, globulin and total bilirubin of liver biomarkers and urea, and creatinine of kidney biomarkers were significantly (p<0.05) elevated in alloxan-induced diabetic rats compared to control group.

AST, ALT, ALP, albumin, globulin, and total bilirubin of liver biomarkers, and urea and creatinine of kidney biomarkers were significant reduced after treatment with A. Occidentale Ethanolic leaves extract, complex extract and metformin compared with the untreated diabetic group. Complex extracts high dose (600 mg/kgb.wt) lowered the markers considerably (Tab. 3a & 3b.) [7-9].


GROUPS
Liver markers Parameters              
Control Diabetic (untreated) Low Extract (400mg/kgb.wt) High Extract (600mg/kgb.wt) Low Complex (400mg/kgb.wt) High Complex (600mg/kgb.wt) Metformin (500mg/kgb.wt)
Albumin (g/dl) 1.74 ± 0.07 2.24  ± 0.81* 2.02  ± 0.20# 1.72 ± 0.05# 1.64 ± 0.06# 1.62 ± 0.06# 1.59 ± 0.06#
Globulin (g/dl) 1.31 ± 0.11 2.23  ± 0.09* 1.47 ± 0.09# 1.39 ± 0.15# 1.36 ± 0.02# 1.24 ± 0.07# 1.32 ± 0.12#
ALP (U/L) 24.66 ± 0.74 36.06  ± 5.59* 26.16  ± 2.03# 23.89 ± 2.16# 24.90 ±1.06# 24.70 ± 1.91# 24.18 ± 0.92#
AST (U/L) 35.26 ± 2.64 75.09  ± 3.52* 39.82  ± 1.15# 35.39 ± 3.38# 37.79 ±7.05# 29.30 ± 2.30# 29.82 ± 0.76#
ALT (U/L) 31.59 ± 1.55 56.62  ± 6.26* 37.06 ±1.56# 35.36 ± 2.56# 37.62 ±3.10# 27.81±2.28# 28.63 ±4.28#
Total bilirubin  (mg/dl) 1.55 ± 0.07 2.68  ± 0.38* 2.06 ±0.20# 1.59 ± 0.18# 2.01 ±0.29# 1.41±0.08# 1.56 ±0.23#

Tab.3a. Effects of A. Occidentale Ethanolic leaves extract and complex extract on liver function markers in alloxan-induced diabetic rats. Values are expressed as mean ± SEM (n=8). *significant at p<0.05 compared with control; #significant at p<0.05 compared with untreated diabetic group.


GROUPS
Peak Number Complex (wave number cm-1) Ligand (wave number cm-1) Band Assignments
1 3641.6 3641.6 O-H Stretching alcohol
2 3391.9 3429.2 N-H Stretching primary amine
3 2288.6 2288.6 C≡N Stretching of nitrile
4 2109.7 2210.2 C≡C Stretching of alkyne
5 1979.2 1982.9 C=C Stretching of allene
6 1852.5 1889.8 C-H Bending of aromatic
7 1781.7 1798.6 C=O Stretching of phenyl esters
8 1654.9 0 C=C Stretching of alkene
9 1476 1397.8 C-H Bending of alkene
10 1416.4 0 O-H Bending of alcohol
11 1062.3 1088.4 C-O Stretching of primary alcohol
12 857.3 0 C-Cl Stretching of an halo compound

Tab. 3b. Effects of A. occidentale ethanolic leaves extract and complex extract on kidney function markers in alloxan-induced diabetic rats.

The spectral are interpreted considering few main peaks observed. The results of the infrared spectra with those of complex and ligand (Tab. 4.). The absorption bands at 3641.6 cm-1 and 3641.6 cm-1 is due to O-H Stretching alcohol. The absorption band at 3391.9 cm-1 in complex due to (N-H) stretching vibration has been shifted to 3429.2 cm-1 in the ligand. The bands at 2288.6 cm-1 in complex, 2288.6 cm-1 were assigned to (C≡N) stretching vibration. The absorption bands at 2109.7 cm-1 increase to 2210.2 cm-1 due to C≡C Stretching of alkyne vibration, The absorption band at 1979.2 cm-1 in complex shifted to 1982.9 in the ligand due to (C=C) Stretching of allene vibration. The bands at 1852.5 cm-1 in the complex increases to 1889.8 cm-1 in C-H Bending of aromatic vibration. The band in the complex at 1781.7 cm-1 shifted to 1798.6 cm-1 at (C=O) Stretching of phenyl esters. There is an absence of absorption band in the ligand, complex with 1654.9 cm-1 in the (C=C) Stretching of alkene. The absorption of band in the complex which decreases from 1476.0cm-1 to 1397.8 cm-1 in the (C-H) Bending of alkene vibration. There is an absence of absorption band in the ligand, complex with 1416.48 cm-1 in the (O-H) Bending of alcohol. The absorption band at 1062.3 cm-1 in complex due to (C-O) Stretching of primary alcohol vibration has been shifted to1088.4 cm-1 in the ligand. There is an absence of absorption band in the ligand, complex with 857.3 cm-1 in the (C-Cl) Stretching of a halo compound [10-14].


Kidney markers Parameters
Control Diabetic (untreated) Low Extract (400mg/kgb.wt) High Extract (600mg/kgb.wt) Low Complex (400mg/kgb.wt) High Complex (600mg/kgb.wt) Metformin (500mg/kgb.wt)
               
Urea (mg/dl) 3.96 ± 0.21 6.33  ± 1.47* 4.10 ± 0.29# 4.02 ± 0.19# 4.15 ± 0.09# 3.81 ± 0.25# 3.84 ± 0.26#
Creatinne (mg/dl)              
0.57 ± 0.02 1.23  ± 0.08* 0.67 ± 0.03# 0.63 ± 0.05# 0.59 ± 0.09# 0.52 ± 0.23# 0.53 ± 0.05#

Tab. 4. Infrared spectral data of ligands and their metal complexes.

DISCUSSION

In this study, anti-diabetic efficacies of A. Occidentale Ethanolic leaves extract and extract complex copper (II) in alloxan monohydrate-induced diabetic rats model were investigated.

Severe body weight loss is a critical symptom of diabetes mellitus. The disparity in energy utilization and expenditure leads to body weight alteration. The loss of body weight might be the result of a decline in glucose metabolism, elevated lipolysis, or structural proteolysis that generates an alternate source of energy. The findings of this study came in line with the result of Faried and El-Mehi, the diabetic rats exhibited a noticeably decrease in body weight. However, administration of the A. Occidentale Ethanolic leave extract and the complex extract caused increase in body weight of the diabetic rats. The observed obvious increase in body weight of diabetic rats treated with the highest Ethanolic extract dose (600 mg/kgb.wt) possibly resulted from its efficient hypoglycemic properties via inhibition of hyperglycemia-induced proteolysis and stimulation of insulin secretion and production to activate fat storage hormone which could be attributed to its bioactive compounds and agreed with the report of Bekara et al [15-19].

Hyperglycemia, the common feature for both type-1 and type-2 diabetes mellitus leads to the pathogenesis of many diabetic complications. In the current study, alloxaninduced diabetic rats showed dramatic elevated blood glucose levels, which correspond with the findings of Arika et al. Elevated blood glucose in diabetic condition may be a consequence of insulin secretion deficiency from pancreatic beta-cells/insensitivity of peripheral tissue to insulin action for glucose uptake and increase hepatic gluconeogenesis. The Ethanolic extract and complex extract administrations effectively lowered the fasting blood glucose levels of the diabetic rats. The complex extract high dose (600 mg/ kgb.wt) and Ethanolic leaves extract high dose (400 mg/ kgb.wt) efficiently reduced the elevated fasting blood glucose levels than other extracts and metformin. Bioactive compounds in medicinal plants have been reported to play a significant role in lowering blood glucose. Hypoglycemic effect of these extracts may be attributable to the presences of bioactive compounds such Phenolics and flavonoids, capable of stimulating insulin secretion from pancreatic beta-cells, facilitate glucose uptake at peripheral tissues, inhibition of hepatic and muscle glucose production, and inhibition of intestinal ⍺-amylase and ⍺-glucosidase enzyme activities and is in line with the report of Shobha and Andallu, on hypoglycaemic effect of anise extract [20-24].

Dyslipidemia, elevated total cholesterol (TC), triglyceride (TG), and altered lipoprotein components noticed by high concentration of low-density lipoprotein-cholesterol (LDL) and low concentration of high-density lipoprotein cholesterol (HDL), is a recognized metabolic abnormality in diabetic condition. Elevated LDL concentration has been linked with progression of cardiovascular disease in diabetic patients. High TC, TG, LDL concentration with diminished HDL concentration remarkably observed in the diabetic rats, consistent with findings of Miaffo et al. Abnormal increment in lipid levels may likely due to deficiency in anti-lipolytic action of insulin to inhibit hormone-sensitive lipase leading to increased mobilization of free fatty acids and peripheral fat deposits. Treatment of diabetic rats in this study with A. Occidentale leaves Ethanolic extract and complex leaves extracts attenuated the dyslipidemia by lowering the TC, TG, LDL levels and ameliorate the HDL level and the complex extract high dose (600 mg/kgb.wt) produce considerable anti-dyslipidemic effect by raising the HDL level almost the same as control level, thereby prevent cardiovascular disease development. Several authors have reported the lipid lowering effect of some bioactive compounds include saponins, phenolic and flavonoids. Attenuation of dyslipidemia in the diabetic rats may also relate to the presence of these bioactive compounds, consistent with report of Limaye et al [25-29]. Diabetic related complications progression has been linked with overwhelming antioxidants enzymes by reactive oxygen species (ROS) generated by oxidative stress. The diabetic rats in this study demonstrated increased oxidative stress marker malondialdehyde (MDA) level and depletion in the activities of antioxidants enzymes superoxide dismutase (SOD), catalase and reduced glutathione (GSH), came in line with findings of Yazdanimehr and Mohammadi. The activities of antioxidants SOD, CAT and GSH were ameliorated with suppressed MDA level in the diabetic rats treated with Ethanolic leaves extract and complexed extract. This revealed its free radical scavenging efficacies via its antioxidant properties; support the findings of Hroob et al. Complex extract lowest dose (400 mg/kgb.wt) strongly boost the antioxidant enzymes activities. Also, phytochemicals flavonoids and phenolics have been identified with antioxidant properties for oxidative stress-related diabetic complications therapy. The presence of these phytochemicals in A. Occidentale leaves may responsible for its antioxidants property [30-34].

In diabetic condition, elevation in hepatic aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase, (ALP) are good indicators for hepatic damage. Also, rise in urea and creatinine signify renal malfunction. Hepatic biomarkers AST, ALT, ALP and renal urea and creatinine were elevated in the diabetic rats of current study, which corroborate with the findings of Wen et al. Oxidative stress, induced many organs complications in diabetic state. The hepatic and renal dysfunction observed might be consequence of excessive free radicals. Elevated AST, ALP, and ALP hepatic markers and renal urea and creatinine levels were restored after administration of A. Occidentale leaves Ethanolic extract and complexed extract to the diabetic rats. Complex extract high dose (600 mg/kgb.wt) appreciably restored the renal and hepatic functions. This suggests that the extracts prevent toxic effect of oxidative stress on organs via its antioxidant property from the bioactive components; concurrence with report of D’Souza et al on amelioration of diabetic nephropathy by Acacia catechu leaves extract [35-41].

CONCLUSION

From this finding, the complexed extract remarkably attenuate hyperglycemia, dyslipidemia and oxidative stress. It also ameliorates the antioxidants activities and protects kidney and liver damages. Complexation of A. Occidentale leaves with copper would improve its potency as novel therapeutic for diabetes mellitus.

DECLARATIONS

Authors’ Contributions

FO, AO conceived the original idea, designed and supervised the research. DE, MO performed the experiment with the support of FO and AM. DE collected the data. DE, FO analyzed the data and prepared the manuscript. FO, MO reviewed the manuscript. All authors’ have read and approved the final manuscript.

AVAILABILITY OF DATA AND MATERIALS

All data generated and analyzed during this study are included in this article.

COMPETING INTERESTS

No competing interests.

FUNDING

No financial fund support received.

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