Protective effects of

Aloe vera

extract on aluminium

sulphate induced alterations in serum lipid pro le of

male albino rats,

Rattus norvegicus

Gajendra Mahor and Sharique A. Ali*

Rajeev Gandhi National Fellow UGC, New Delhi Department of Biotechnology, Sai a Science College,

Bhopal - 462001, India

ABSTRACT

The present study suggests that treatment with Aloe vera a medicinal plant belonging to the family - Liliaceae, used

in traditional Indian medicine system and its active constituent Aloin has a positive and therapeutic effect in lowering

the lipid pro le level in aluminium sulphate exposed rats for a period of 60 to 90 days. Lipid pro le (total cholesterol,

triglyceride, HDL and LDL) levels were found to signi cantly increased (P<0.05) after treatment of Al

(So

)

in Group

II compared to normal control Group I treated with normal diet. Group III and Group IV animals treated with Al

(So

)

and Aloe vera extract and Al

(So

)

and Aloin respectively, showed signi cant decrease in lipid pro le at (P<0.05).

The present study also validates that A.vera extract and pure aloin was effective in reducing Al toxicity in lipid pro le

(Total Cholesterol, Triglyceride, HDL and LDL) of treatment in the long term 60 and 90 days of aluminium exposed rats.

KEY WORDS:

ALOE VERA

, ALUMINIUM TOXICITY, TOTAL CHOLESTEROL, TRIGLYCERIDE, HDL AND LDL

727

Toxicological

Communication

Biosci. Biotech. Res. Comm. 11(4): 727-733 (2018)

INTRODUCTION

Aluminium (Al) is the third most abundant metal present

naturally in the Earth’s crust.

It is also present in soil,

air, water, several eatables, and commercial products

such as food storage material, cookware, and medicinal

products including drugs. Exposure to humans occurs

through different routes. The common routes of expo-

sure include inhalation, oral, and skin. Exposure is more

common among people working in Al industries. The

extensive use of Al cookware leads to ingestion of small

quantities of Al every day. Al is found to be a compo-

nent of commonly used medications such as anti-ulcer

drugs such as sucralfate, antacids containing Al, hae-

ARTICLE INFORMATION:

Corresponding Authors: drshariqali@yahoo.com

Received 19

Sep, 2018

Accepted after revision 21

Dec, 2018

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007 CODEN: USA BBRCBA

Thomson Reuters ISI ESC / Clarivate Analytics USA

Mono of Clarivate Analytics and Crossref Indexed

Journal Mono of CR

NAAS Journal Score 2018: 4.31 SJIF 2017: 4.196

Online Contents Available at: http//www.bbrc.in/

DOI: 10.21786/bbrc/11.4/25

Gajendra Mahor and Sharique A Ali

728 PROTECTIVE EFFECTS OF

ALOE VERA

EXTRACT ON ALUMINIUM SULPHATE INDUCED ALTERATIONS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

modialysis  uid, phosphate binders and vaccines.

Al is

also found in anticaking agents, preservatives,  llers,

coloring agents, emulsi ers and baking powders.

Such

extensive use of Al in consumable and non consumable

products will certainly lead to Al entry and deposition

in human body, (Denise etal., 2007; Verstraeten etal.,

2008; Gura, 2010; Thirunavukkarasu etal., 2013; Kalai-

sevi etal., 2015; Sakr etal., 2017; Konda etal., 2017;

Ahmed etal., 2018).

Al does not have any physiological role in the body

but it gets stored mainly in the blood, lungs, liver,

bones, brain, spleen, kidney and muscles. It may act

as a competitive inhibitor for elements such as magne-

sium, iron and calcium because of its atomic size and

electric charge and may results in anaemia and bone

damage. Al-induced neurotoxicity and changes in serum

lipid pro le and vitamins. High level of exposure can

cause toxicity such as nephrotoxicity and hepatotoxic-

ity. It was already been reported in patients with chronic

kidney disease who were on dialysis with Al-containing

dialysis  uid.

Al toxicity has been associated with Alz-

heimer’s disease, dialysis, Parkinson’s dementia. It is due

to oxidative stress and lipid peroxidation in tissues, Pro-

tein and DNA (Tchounwou etal., 2012; Thomford etal.,

2017; Azza etal., 2017).

Lipid is an important component of human body

because it is a main constituent of cell membrane, sev-

eral hormones and also performs many other cellular

functions (Esther et al., 2013). Lipids being insoluble

in the blood so it is transported from the cells by low

density and high density lipoproteins (Brown et al.,

2007; Kaji etal., 2013). High density lipoproteins (HDL)

tend to carry cholesterol away from arteries back to the

liver (Van der Veen etal., 2009). Therefore, high serum

cholesterol level can be due to hepatic dysfunction.

Although several factors, such as life style, a diet rich

in cholesterol, age and hypertension, have been reported

to cause heart failure (Kumar etal., 2011). High levels

of cholesterol, particularly LDL cholesterol, are mainly

responsible for hypercholesterolemia provoked cardio

toxicity (Azad etal., 2001).

Several anti-hyperlipidemic agents are currently

available; however most of them have associated with

various unwanted effects. Hence, people are switching

towards safer alternatives, specially derived from plants

with limited side effects. The World Health Organization

(WHO) has given its estimation that more than 2/3

the global population in recent times depends on alter-

native sources of treatment to ful l the basic health care

requirements and this most importantly embroils the

usage of plant products. This means that nearby two-

thirds of the people globally trust on plants as a reliable

way of their medication. Nowadays, vigorous research

is ongoing to discover nontoxic and bene cial herbs.

Herbal Medicine or herbalism is the practice or art of

employing herbs and herbal preparations in order to

remain healthy and also for the treatment and improve-

ment in prognosis of diseases. A.vera is a medicinal

plant belongs to the family Liliaceae

its active constitu-

ent aloin have antioxidant properties, protective against

heavy metal toxicity. Its therapeutic applications include

wound healing, diabetes, burns for easing intestinal,

curing ulcers and arthritic swelling (Kumar etal., 2010;

Sai etal., 2011; Jakkala etal., 2015; Mahor etal., 2016;

Gupta etal., 2017). The aim of the present study was to

investigate the protective role of A.vera on Al induced

changes in lipid pro le (cholesterol, triglyceride, HDL

and LDL) of experimental rats.

MATERIALS AND METHODS

Collection and identi cation of plant material: The

fresh leaves of A.vera (Aloe barbadensis) were collected

from the Minor Forest Produce Processing and Research

Centre (MFP-PARC) Van Parisar, Barkhera Pathani, Bho-

pal, (M.P.) India. The plant was authenticated by Dr.

Zia-Ul-Hassan Head of the Department of Botany at the

Sai a College of Science Peer Gate, Bhopal, (M.P.) India

and the voucher specimen (403/Sai a/Bot/16) has been

deposited at the Herbarium of the Sai a Science College,

Peer Gate, Bhopal, (M.P.) India.

Preparation of extracts: After collection and weigh-

ing, fresh leaves of Aloe vera were washed with distilled

water to remove dirt and dried under shade separately.

The extraction of A. vera leaves was done according

to the method (Kumar & Muthuselvam, 2009). Slight

modi cation, Skin of the leaves were pealed and the

gel inside was used for extraction. 100 gm of the gel

was added to 250ml of ethanol and extracted using the

Soxhlet assembly. Later on, the solvent of the extracted

material was removed at low temperature in a rotary

vacuum evaporator and the resulting dried extract was

lyophilized in a freeze dryer.

Drugs and chemicals: In this study, Al-sulphate (Al

)

was purchased from Aldrich chemical Company (St.

Lousis mo, USA) and Standard Aloin (C21H22O9) was

obtained from Sigma. The diagnostic kits required for

enzymatic assays were purchased from Span Diagnos-

tics. All other chemicals used in the experiment were

of analytical grade. The dose of Al-sulphate (Al

)

was 98mg (Al

)

/L (1/25 LD50). The dose of A.vera

extract and Aloin were 100 mg/kg BW. These doses were

selected based on basis of pilot experiments.

Maintenance of animals and approval of proto-

col: Healthy adult male albino rats (Rattus norvegicus)

weighing 120-150g were used for the present investiga-

Gajendra Mahor and Sharique A Ali

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS PROTECTIVE EFFECTS OF

ALOE VERA

EXTRACT ON ALUMINIUM SULPHATE INDUCED ALTERATIONS 729

tion. They were housed in a clean polypropylene cage

and maintained in an air-conditioned experimental

room at 12-hour light: dark cycles. The animals were

acclimatized to laboratory condition for one week

prior to experiment. Standard pellets were used as a

basal diet during the experimental period. The control

and experimental animals were provided with puri ed

drinking water ad libitum. The animals were main-

tained in accordance with the “CPCSEA guidelines for

laboratory animal facility” (Committee for the Purpose

of Control and Supervision on Experiments on Ani-

mals) and the approval number is CPCSEA Registra-

tion number SSC/06-06-22/CPCSEA, dated 26/10/2006.

Before starting the experiment the animals were care-

fully marked on different parts of their body, which

was later used as identi cation mark for a particu-

lar animal, so that the response of a particular mouse

prior to and after the administration could be noted

separately.

Acute oral toxicity studies: A.vera extract at the dose

range of 100–2500 mg/kg body weight were adminis-

tered by oral gavage method on different group of mice

comprised of 6 rats in each group. Animals were kept

under close observation for 4 hours after administer-

ing the fraction for behaviour, neurological, and auto-

nomic pro le and then observed for any change in the

general behaviour and physical activities; mortality was

recorded within 72 hours. Acute toxicity was determined

according to the method (Lorke, 1983).

Induction of Toxicity/experimental design: A total of

24 male (2 months old) Albino rats (Rattus norvegicus)

weighing 120-150g were used for the present investiga-

tion. The animals were divided into four groups (6 rats/

group): Group I:-was kept as control without giving any

treatment. Compared to adult controls, Group II: - ani-

mals in this group were given 17±6 ml of water sup-

plemented with Al-sulphate to consume, corresponding

to 98 mg of Al per day (Laxman etal., 2016) for 60 and

90 days. Group III: - This group animals were fed with

normal diet and received aluminium sulphate (98 mg/

kg body weight) and Aloe vera extract (100 mg/kg body

weight) for 60 and 90 days. Group IV: - This group ani-

mals were fed with normal diet and received aluminium

sulphate (98 mg/kg body weight) and Aloin (100 mg/kg

body weight) for 60 and 90 days.

Animal Grouping and Treatment Schedule: Four

groups of rats, six rats in each, received the following

treatment schedule: Group I rats received normal diet

and water ad libitum, as control group. Group–II rats

administered twice with Aluminium sulphate (98 mg/

kg/day) dissolved in (1ml/kg b.wt) water were injected

dose orally for 60 and 90 days. Group III, will be admin-

istered with Aluminium sulphate (98 mg/kg/b.w.) with

Aloe vera extract (100 mg/kg/b.w.) dose orally for 60 and

90 days and last Group-IV rats were administrated Alu-

minium sulphate (98 mg/kg/b.w.) with Aloin (100 mg/

kg/b.w.) dose orally for 60 and 90 days.

Collection of Blood Sample and Estimation of Serum

Lipid pro le Investigations: Blood samples were col-

lected by orbital sinus puncture method (Hui et al.,

2007). Serum was separated by following procedure.

Blood samples were withdrawn from orbital sinus using

non heparinised capillary tubes, collected in dried cen-

trifuge tubes and allowed to clot. Serum was separated

from the clot by centrifuged at 3000 rpm for 15 min.

at room temperature. Serum was collected carefully and

kept at-20°C until analysis of Total cholesterol, High

Density Lipoprotein (HDL) cholesterol and triglycerides

by using kits supplied by Span Diagnostic Ltd. Plasma

concentrations of total cholesterol, triglycerides, HDL &

LDL fractions were measured by using standard methods

with commercially available kits. LDL cholesterol was

calculated with the Friedewald formula as follows: LDL

cholesterol = total cholesterol − HDL cholesterol − (tri-

glycerides/5) (Friedewald etal., 1972).

STATISTICAL ANALYSIS OF DATA: All parameters

were presented as mean ± SEM. One-way analysis of

variance followed by Bonferroni multiple comparisons

using a computer-based  tting program (Prism, Graph

Pad) were performed. Differences were considered to be

statistically signi cant when P < 0.05.

RESULTS

It was obsevered that all four groups of rats received

the following treatment schedule: shows the signi cant

change in all parameters discussed here. After 60 days

(Group II) showed a signi cant (P<0.05) increase in the

level TC, TG, HDL and LDL due to Al toxicity compared

to group I. whereas signi cant (P<0.05) decrease in TC,

TG, HDL and LDL level was reported in group III and

group IV (Table:1), (Fig: 1).Experimental results shows.

After 60 days group III and group IV showed a signi -

cant (P < 0.05) decrease in the level TC, TG, HDL and

LDL which is induced due to Al toxicity group II com-

pared to group I (Table: I), (Fig: I).

After 90 day study it was observed that Al toxicity

enhances compared to 60 or 90 days. It means Al on

long term exposure induces toxicity in group II whereas

A.vera extract and aloin was also effective in reducing

toxicity in various parameters studied after 90 days.

After 90 days group III and group IV showed a signi -

cant (P<0.05) decrease in the level of TC, TG, HDL and

LDL which is induced due to Al toxicity group II nearest

about to group I. (Table:II) and (Fig: II),

Gajendra Mahor and Sharique A Ali

730 PROTECTIVE EFFECTS OF

ALOE VERA

EXTRACT ON ALUMINIUM SULPHATE INDUCED ALTERATIONS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

FIGURE 1. Graph showing variation in different levels of Total Cholesterol: Triglycerides, HDL

and LDL against Aluminium sulphate (98mg/kg/bw) induced toxicity after 60 days.

Table 1. Effects of orally administrated A. vera extract and aloin on Total Cholesterol, Triglycerides, High

density lipoproteins, Low density lipoprotein intoxicated with Aluminium sulphate after 60 days.

Group Treatment

Total Cholesterol

(mg/dl)

Triglycerides

(mg/dl)

HDL (mg/dl) LDL (mg/dl)

I Normal Control 99.33±5.391 93.33±5.755 38.17±5.115 31.50±1.871

II Al2(So4)3 (98mg/kg) 129.83±2.316 139.83±3.312 17.83±2.317 50.67±4.179

III

Al2(So4)3 (98mg/kg) +

A.vera extract (100gm/kg)

95.16±2.136 * 95.17±4.446* 33.17±5.037* 27.50±4.848 *

Al2(So4)3 (98mg/kg) +

Aloin (100gm/kg)

95.33±3.444** 87.83±4.401** 37.33±5.715 ** 25.17±2.317 **

* & **= indicates signi cant values, signi cantly different at P  0.05.

Table 2. Effects of orally administrated A. vera extract and aloin on Total Cholesterol, Triglycerides, High

density lipoproteins, Low density lipoprotein intoxicated with Aluminium sulphate after 90 days.

Group Treatment

Total Cholesterol

(mg/dl)

Triglycerides

(mg/dl)

HDL (mg dl) LDL (mg/dl)

I Normal Control 101.83±4.622 94.67±5.785 40.00±3.950 33.17±2.927

II Al2(So4)3 (98mg/kg) 133.17±4.070 141.17±4.834 15.67±1.633 53.67±4.633

III

Al2(So4)3 (98mg/kg) +

A.Vera extract (100gm/kg)

92.83±3.189* 92.50±5.925* 35.50±3.507 * 29.50±4.593 *

Al2(So4)3 (98mg/kg) + Aloin

(100gm/kg)

92.67±8.238 ** 85.50±9.731 ** 38.67±4.885 ** 23.67±3.983 **

* & ** = indicates signi cant values, signi cantly different at P  0.05.

Gajendra Mahor and Sharique A Ali

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS PROTECTIVE EFFECTS OF

ALOE VERA

EXTRACT ON ALUMINIUM SULPHATE INDUCED ALTERATIONS 731

DISCUSSION

The hypertriglyceridemia and lipid oxidation were main

features of this altered metabolism. Hyperlipidemia is a

condition where there is an elevation of the serum levels

of total cholesterol (TC) and triglycerides (TG) due to the

lipid metabolism alteration, with an increase in the liver

lipogenesis and lipolysis in the adipocytes. Low-den-

sity lipoprotein (LDL) is the compound containing both

lipid and protein, which transport cholesterol to tissues

other than the liver. High-density lipoprotein (HDL) is

the compound containing both lipid and protein, which

transport cholesterol to the liver for excretion in the bile.

(Kalaiselvi etal., 2015; Gouda etal., 2018).

Aluminium (Al) is toxic to humans and animals. Its

toxicity results to generation of reactive oxygen species

in lipids which leads to oxidative damage of biomol-

ecules in an organism. The present study investigated

the effects of Al-sulphate in toxicity induction and

bene cial effect of A.vera, aloin against the induced

toxicity in rats. The  ndings of this study were that Al

perturbed the metabolism of lipids (cholesterol, triglyc-

eride, LDL and HDL) in rat. It may up-/down-regulation

the levels of these lipids due to up- or down-regulated

of enzyme. These perturbations were presented in the

plasma as hypertriglyceridemia, hypercholesterolemia

and hypophospholipidemia. the increase in plasma cho-

lesterol as a result of ingestion of Al.

Due to Al inges-

tion caused a preferential activation of receptor sites

on the cells which favoured the synthesis of cholesterol

in these organs by up-regulating hydroxymethylglu-

taryl coenzyme A reductase (a rate-limiting enzyme in

cholesterol synthesis pathway) since virtually all cells

can synthesize cholesterol or Al changes the integrity

of the cell membrane thereby causing a constipation of

cholesterol in the organs by modi cation of the com-

position, structure and stability of the cell membranes.

The liver has been shown to be one of the target organs

of Al toxicity- induced injury, liver damage is likely to

cause some membrane lipids to be released into circu-

lation; metabolism with oxidative stress and lipid per-

oxidation and reactive oxygen species as hydroxyl and

superoxide radicals in liver alter the lipid level in serum

(Kolomiytseva, 2011; Ugbaja etal., 2015; Younes etal.,

2018).

Al causes toxic effect on biochemical parameters i.e.

Cholesterol, Triglycerides It shows an increasing trend

because prolonged metallic stress in the experimental

animals makes adaptation dif cult and creates weak-

ness, anemia. In the  eld of environmental bio moni-

toring these parameters have been effectively used

as potential biomarkers of Al toxicity in animals and

human. This present study was carried out to investigate

the effect of A.vera, a well-known medicinal plant with

antioxidant properties, on Al-induced alterations in lipid

metabolism. In comparison to controls, rats with Al tox-

icity displayed higher cholesterol, triglyceride, HDL and

LDL concentrations in serum (Joshi etal., 2013).

FIGURE 2. Graph showing variation in different parameter levels of Total Cholesterol:

Triglycerides, HDL and LDL against Aluminium sulphate (98mg/kg/bw) induced toxicity

after 90 days.

Gajendra Mahor and Sharique A Ali

732 PROTECTIVE EFFECTS OF

ALOE VERA

EXTRACT ON ALUMINIUM SULPHATE INDUCED ALTERATIONS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

A.vera extract has a wide range of therapeutic appli-

cations. A.vera gel contains anthroquinones (aloin, aloe-

emodin) which may have a variety of properties of anti-

oxidant agent, including the protective role for heavy

metal toxicity. Previous studies have also shown that

as an antioxidant, plant extracts may improve the pro-

oxidant effects of Al (Nada etal., 2013; Jakkala etal.,

2016; Mahor etal., 2016).

In this study, the A.vera extract proved to be quite

effective in lowering the lipid pro le (total cholesterol,

triglycerides, HDL and LDL) Al toxicity. Al in the blood

was signi cantly reduced due to administration of the

A.vera extract since it possesses chelating properties.

Administration of the A.vera extract for 60 and 90 days

lead to decreases in cholesterol, triglycerides, HDL and

LDL levels in the Al-sulphate exposed animals. This

implies an exacerbating effect of A.vera on Al toxicity.

CONCLUSION

The present study also validates that A.vera extract and

aloin was effective in reducing Al toxicity in lipid pro le

(Total Cholesterol, Triglyceride, HDL and LDL) of treat-

ment 60 and 90 days.

CONFLICTS OF INTEREST

The authors have no con ict of interest to declare.

ACKNOWLEDGEMENT

GM is thankful to UGC New Delhi for awarding RGNF,

Vide File No.2014-15-SC-MAD-67686/ 2014.

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