Accepted Manuscript

Combination effect naringin and pravastatin in lipid profile and

glucose in obese rats

Abdel K. Raffoul-Orozco, Ana E. Ávila-González, Christian M.

Rodríguez-Razón, Teresa A. García-Cobian, Edsaul E. Pérez-
Guerrero, Trinidad García-Iglesias, Edy D. Rubio Arellano

PII: S0024-3205(17)30621-5
DOI: doi:10.1016/j.lfs.2017.11.044
Reference: LFS 15456
To appear in: Life Sciences
Received date: 1 August 2017
Revised date: 12 November 2017
Accepted date: 26 November 2017

Please cite this article as: Abdel K. Raffoul-Orozco, Ana E. Ávila-González, Christian M.
Rodríguez-Razón, Teresa A. García-Cobian, Edsaul E. Pérez-Guerrero, Trinidad García-
Iglesias, Edy D. Rubio Arellano , Combination effect naringin and pravastatin in lipid
profile and glucose in obese rats. The address for the corresponding author was captured
as affiliation for all authors. Please check if appropriate. Lfs(2017), doi:10.1016/
j.lfs.2017.11.044

This is a PDF file of an unedited manuscript that has been accepted for publication. As
a service to our customers we are providing this early version of the manuscript. The
manuscript will undergo copyediting, typesetting, and review of the resulting proof before
it is published in its final form. Please note that during the production process errors may
be discovered which could affect the content, and all legal disclaimers that apply to the
journal pertain.
 ACCEPTED MANUSCRIPT

Combination effect naringin and pravastatin in lipid profile and

glucose in obese rats

Abdel K. Raffoul-Orozcoa, Ana E. Ávila-Gonzáleza, Christian M. Rodríguez-Razónb, Teresa

A. García-Cobiana, Edsaul E. Pérez-Guerreroc, Trinidad García-Iglesiasb, Edy D. Rubio

Arellano*d

PT
a
Institute of Experimental and Clinical Therapeutics, Department of Physiology, Health

RI
Sciences University Center, University of Guadalajara, Sierra Mojada st. 950. P.C. 44340
b
Guadalajara, Jal, Mex 01 33 1058 5200. Laboratory immunology. Department of

SC
Physiology, Health Sciences University Center, University of Guadalajara, Sierra Mojada st.
NU
950 Guadalajara, Jal, Mexico 01 33 1058 5200, c Doctorate of pharmacology. De partment of

Physiology, Health Sciences University Center, University of Guadalajara, Sierra Mojada st.
MA

d
950. P.C. 44340 Guadalajara, Jal, Mexico 01 33 1058 5200, Laboratory Pharmaceutical

Research and Development. Department of Pharmacobiology. Exact sciences and engineering

D

center. University of Guadalajara, Marcelino García Barragan Blvd. 1421 P.C. 44430.
E

Guadalajara, Jal. Mexico 01 33 13785900

PT

*Corresponding Author:
CE

Edy David Rubio-Arellano PhD MD. Laboratory Pharmaceutical Research and Development.

Department of Pharmacobiology. Exact sciences and engineering center. University of

AC

Guadalajara, Marcelino García Barragan Blvd. 1421 P.C. 44430 Guadalajara, Jal. Mexico 01

33 13785900

E-mail address: edy1983@icloud.com

1
 ACCEPTED MANUSCRIPT

ABSTRACT

Aims: The purpose of this study was to compare the effect of naringin 100 mg/kg in

combination with pravastatin 10 mg/kg by gavage for 6 weeks compared with monotherapy

over lipid profiles, glucose levels and weight in murine model of obesity.

Main methods: The study design was planned with 5 groups of 6 male Wistar Albina rats:

Group 1: control with balanced food and vehicle (C-); Group 2: control with Obesity and

PT
vehicle (C+); Group 3: Obesity + naringin (N); Group 4: Obesity + pravastatin (P); Group 5:

RI
Obesity + pravastatin + naringin (NP). Obesity was developed with a food model.

SC
Key findings: The naringin groups showed a decrease in weight gain and low glucose values

compared to the control group (weight NP:311.4 vs C+:348.6; glucose NP: 173.12 vs
NU
C+:235.56) (p <0.05); the group with naringin+pravastatin combination showed the total

cholesterol (TC), LDL and triglycerides (TGs) to normal levels (TC NP:51.6 vs C+:83.4;
MA

LDL NP:9.32 vs C+:32.32; TGs NP:39.4 vs C+:89.4) (p <0.05); but was not statistically

significant compared with monotherapy.

E D

Significance: The combination of naringin and pravastatin did not appear to be better than
PT

monotherapy on lipids, but its use could generate euglycemic and antiobesogenic effects, in

addition to diminishing the adverse hepatic effects of pravastatin in rats.

CE

Key-words: Naringin, pravastatin, phytopharmaceuticals, dyslipidemia, DM2, obesity.

AC

2
 ACCEPTED MANUSCRIPT

INTRODUCTION

Obesity is one of the most prevalent diseases, a growing public health problem in Mexico as

in the world [1,2]. Obesity is related to other metabolic diseases such as dyslipidemia and

type 2 diabetes, among others [3-5]. Although there are drugs for treating each of these

diseases, the use of nutraceuticals or phytodrugs have yielded good results [6-8], and their

combination is a field that has been briefly studied [7-9].

PT
RI
Naringin (chemically 4',5,7-rhamnoglucoside trihydroxyflavanone-7) is a citru-flavanone

SC
glycoside extracted from citrus fruits, although found in greater proportion in the pericarp of

citrus paradisi also used in herbal medicine with good results in murine models and in
NU
clinical trials [10], according to studies, it inhibits the 3-hydroxy-3methylglutaryl CoA

reductase (HMG-CoA), lipoxygenase, and regulates PPAR, among other mechanisms of

MA

action, achieving effect on serum lipids similar to statins, besides in DM2 and anti-obesogenic

effects [11,12], according to experts, its use should not replace statins therapy [9]. Naringin is
D

a product used in the food industry, which is an easy and inexpensive extract to obtain,
E
PT

adhering the therapy for dyslipidemia could be beneficial in lowering treatment costs, also as

an alternative treatment for patients who have not achieved lipid goals, or patients with
CE

intolerance to statins as suggested in the Position Paper from an International Lipid Expert

Panel and in the review study of Robert S. Rosenson et al, in the Journal of the American
AC

College of Cardiology [9, 13].

Statins are inhibitors of HMG-CoA, first choice therapy for dyslipidemia [14,15], pravastatin,

one of the safest even in patients with multiple treatments, being metabolized in the kidney

which does not require hepatic metabolism by the CYP450 family, that has less activity as an

antagonist [13,16].

3
 ACCEPTED MANUSCRIPT

There are several studies with naringin and pravastatin, however, only in one report it

evaluates co-administration, it concluded the safe administration in patients [17]. Until now,

there are no studies evaluating the combination of metabolic diseases, given that the

individual characteristics may result with a decrease of lipid concentration greater than

monotherapy; if results are positive in addition to synergic effect, studies could be carried out

PT
with this combination in patients to evaluate it as an additional tool for the treatment of

RI
dyslipidemia with pleiotropic effects added to statins, especially for patients with obesity or

SC
untreated patients with hyperglycemia.
NU
The purpose of this study was to evaluate the effects of combining pravastatin and naringin in

serum concentrations for lipids and glucose in the obesity murine model.
MA

MATERIAL & METHODS

D

Animals. Thirty male Wistar rats, 5 weeks of age, weighing 100 ± 20 grams, were evaluated;
E
PT

the rats were housed in groups per cage and were kept in well-ventilated animal rooms in a

temperature of 25-27 ° C in a light / dark cycle of 12 hours. Rats had free access to a modified
CE

diet and drinking water ad libitum

AC

Ethical consideration. The animals were handled with the institutional guidelines of the local

Ethics Committee and the Mexican Official Standards (NOM-062-ZOO-1999) the projects

approval number DF/CB068/13, and international guidelines on the Use and Handling of

Experimental Animals [18].

4
 ACCEPTED MANUSCRIPT

Induction of Obesity, Diet-model. Obesity was induced in rats by feeding a hyper caloric

nutritional model (diet-model obesity DMO), which was taken as the food base, “Purine,

Rodent Chow®”, immersed for 10 minutes in lard, which was previously melted at 90 ° C,

then left at room temperature for easy manipulation, enough sucrose (10% dry weight) to

cover the food completely, a homogeneous mixture was added. Food was offered on free-

demand the model was reached after 8 weeks, which reached a higher total body weight to a

PT
normal body weight (>50% total body weight) [19]; in rats with a balanced diet (Rodent

RI
Chow®) the animal reached an average weight (200±20 g) which was used as a negative

SC
control. Nutritional data is shown in Table 1.
NU
TABLE 1. Nutritional information. Comparison of calories and percentages between balanced food

and modified food for the obesity model

MA

RODENT CHOW DIET, PURINA®

Standard Hyper caloric

D

CALORIC DENSITY 3.1 kcal/g 5.9 kcal/g

E

PROTEINS 18% 14%

PT

LIPIDS 24% 38%

CE

CARBOHYDRATES 58% 48%

Kcal: kilocalorie, g:grams,

AC

Treatment. The animals were randomly divided into five groups. Treatment was administered

by gavage. Vehicle: saline solution, naringin (100 mg / kg/ d) and / or pravastatin (10 mg / kg/

d) once daily between 08:00-09:30 am for 6 weeks.

Group 1: Control with balanced food and vehicle, ie normo weight (C-) (note: group used

only to define the presence of the disease in the murine model); Group 2: control with DMO

5
 ACCEPTED MANUSCRIPT

and vehicle, ie control obese group (C+) Group 3: DMO + naringin (N); Group 4:

DMO+pravastain+naringin; (NP) Group 5: DMO+pravastatin (P); note: pravastatin was

administered 1 hour before naringin.

After completing a 6-week intervention, the rats fasted for 12 to 16 hours. The animals

were sacrificed under anesthesia, the blood sample was taken by cardiac puncture

approximately 5 to 8 ml were obtained. Blood samples were collected, centrifuged at 1500

PT
rpm (600 x g approx.) for 10 min, the supernatant (serum) was collected individually and

RI
stored at -20°C until analysis.

SC
Intervention. Naringin was acquired from "SAINA Xi'an Biological Technology Co., Ltd."
NU
China, with high degree of purity, concentration of 98%. Quantified by HPLC. Naringin was

dissolved by stirring in saline solution, counted in adequate doses per ml.

MA

Pravastatin used was Novina® 30 tablets of 10 mg. Novag Laboratories (Novag Infancia, S.A.

de C.V.). Pravastatin was pulverized and administered in sufficient amounts per rat in 1 ml
D

saline solution.
E

Biochemical Analysis. The serum concentrations of Total cholesterol (TC), triglycerides

PT

(TGs), high density lipoprotein (HDL) cholesterol, glucose, ALT, AST and creatinine for
CE

biosafety were determined by using the enzyme-linked colorimetric technique. Serum

samples were removed at a temperature of -20° C then used at room temperature; the
AC

calculation of the results were done with an automated software ERBA 1100

spectrophotometer.

Serum low density lipoprotein (LDL) cholesterol was estimated using the equation by

Friedewald [20], and used bay Erejuwa and cols [21,22].

6
 ACCEPTED MANUSCRIPT

Statistical Analysis. Was performed using the Kruskal-Wallis test to compare results between

groups. Post hoc testing was used, the U of Mann-Whitney test. In addition to the comparison

of changes in body weight using the Wilcoxon test for intergroup testing before and after the

intervention. p ≤ 0.05 was considered a statistical significance.

The results were analyzed using SPSS version 21. Data are expressed as mean ± SD.

PT
Diet-model obesity. A comparison of the results between the group of normal weight “control

RI
(-)”, and obese rats untreated, “control (+)” was performed, this is to show model results and

SC
define the presence of the disease being treated. Table 2 shows significant statistical and

clinical differences between the main parameters, getting rats weighing more than 50% of
NU
their weight than the normal weight rats (p = 0.008) in groups before the beginning and the

end of the intervention, plus the presence of hyperglycemia comparing observed the values
MA

obtained in the normal weight rats against the values in obese rats, finally altered lipid values

were obtained by total cholesterol, triglycerides and LDL cholesterol.

E D
PT
CE
AC

7
 ACCEPTED MANUSCRIPT

TABLE 2. Results of diet-model obesity. Model of obesity, hyperglycemia and high

serum lipids in rats are shown. Weight expressed in grams, the rest of the values expressed

in mg/dl

Control normo-weight Control obese p=

mean ± SD mean ± SD

Baseline Weight 193.60 ± 8.8 306.40 ± 6.2 0.01**

PT
Final Weight 223 ± 23.9 348.60 ± 37.7 0.01**

RI
Glucose 99.5 ± 11.5 235.6 ± 77.9 0.01**

SC
Total Cholesterol 53.54 ± 2.8 83.4 ± 12.3 0.01**

LDL-C 7.96 ± 5.3 32.32 ± 20 0.05*

NU
HDL-C 37.98 ± 6.2 41.4 ± 6.8 0.31

Triglycerides 38 ± 7.5 89.4 ± 21.8 0.01**

MA

Creatinine 0.68 ± 0.08 0.8 ± 0.2 0.31

ALT 53.47 ± 6.5 50.36 ± 7.2 0.73

D

AST 142.6 ± 22.5 172.5 ± 47.8 0.42

E

.* refers to p≤0.05, ** p=≤0.01.

PT
CE

Biosafety in diet-model obesity. To assess the biosafety of the disease model, renal creatinine

was measured, as well as alanino aminotransferase (ALT) and aspartate aminotransferase

AC

(AST) for liver biosafety; there were no clinical differences or statistics, p > 0.05, dates are

shown in table 2.

RESULTS

Effect of naringin and/or pravastatin in Body Weight (BW) of obese Rats. The data of the

weight at the beginning and at the end of the intervention are shown in Table 3. There were

8
 ACCEPTED MANUSCRIPT

no indicators of a significant difference between groups of obese rats. The comparison

between “” of each intervention group and control (+) was performed, the results are shown

in Figure 1, in which naringin shows a statistically significant difference in p = <0.05

compared to the control (+) and obese rats treated with pravastatin.

TABLE 3. Basal and final weight of obese rats. The mean and SD obtained from the total

PT
body weight in grams.

RI
Group Average basal weight Average final weight p=

SC
Control (+) 306.4±6.2 348.6±12 0.01*

Naringin 294.6±4.3 306.6±6.1 0.01**

NU
Naringin + pravastatin 293.8±6.8 311.4±6.6 0.01**

Pravastatin 303.8±8.8 330.6±23.9 0.01**

MA

* refers to p≤0.01 compared between basal vs. final weight, add* refers to p≤0.01 compared

with control (+) final weight.

E D
PT
CE
AC

9
 ACCEPTED MANUSCRIPT

Effects of naringin and/or pravastatin on Blood Glucose (BG) Concentrations. Figure 2

shows the effect of naringin and/or pravastatin on glucose concentrations. The concentrations

obtained by the diet-model are displayed in the control (+) group, we can observe that the

naringin group obtained the best results (p <0.05), although the other 2 treatment groups

showed similar values, that were not statistically significant.

PT
RI
SC
NU
MA
E D
PT
CE

Effects of naringin and/or pravastatin on triglycerides (TGs) total cholesterol (TC), High
AC

Density Lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) of

obese Rats. The data on the effects of naringin and/or pravastatin on TGs, TC, HDL and LDL

in obese rats are represented in Figure 3. In the comparative analysis of TGs in groups, a

statistically significant difference p = 0.003 was found, post hoc tests were performed, we

found a significant difference in all treatment groups vs. control (+), resulting p = 0.009 (*), in

addition, we found that the naringin group also has a significant difference compared to the

10
 ACCEPTED MANUSCRIPT

pravastatin group (**). The results of total cholesterol show a statistical significance of p =

0.02, performing post hoc tests, finding that the groups tested showed a difference against the

control group (+), with p = <0.05.

In the analysis of missing lipids, ie, HDL and LDL cholesterols, there were no statistically

significant differences, although in the comparison group tested against the control group (+)

it could be observed with clinically significant differences in LDL, which is shown in the

PT
figure 3.

RI
SC
NU
MA
E D
PT
CE
AC

Biosafety treatments. Figure 4 shows the biosafety of the various treatments used in addition

to being contrasted with the control (+). In the pravastatin group shows ALT elevated which

is statistically significant, not clinically. The naringin group pravastatin did not show the

elevation of ALT that produced in pravastatin as monotherapy, the combination not only

showed safety but also decreased the adverse effects of pravastatin.

11
 ACCEPTED MANUSCRIPT

PT
RI
SC
NU
MA

DISCUSSION

A high-calorie food, high level consumption of sucrose model for this research was used
E D

[19,23] according to the results we found its feasible use and reproduction to establish a
PT

model with diseases linked to obesity, hyperlipidemia with elevated levels of TGs, total

cholesterol, LDL and hyperglycemia, although showing different glucose levels, high values
CE

were presented in all experiment rats, which helped us evaluate interventions to measure a
AC

hypoglycemic or euglycemic effect. Compared to other models used, this study was

inexpensive and did not require chemical additives, making it a safer model, but may present

variation in respect to models enriched with lipids as cholesterol directly added or toxic

agents, such as alloxan and estreptosotozin models [21,22,24]; although they generate

excellent results to simulate a DM2 there may be side effects and not show obesity.

12
 ACCEPTED MANUSCRIPT

The doses of naringin and pravastatin for the present study were taken from studies conducted

in murine models that showed desired effects, besides showing the peaks of absorption and

bioavailability to determine the time of interventions, particularly in combination [25,26].

The results indicated that administration of naringin generates a decrease in weight gain in

the diet model used, this is reflected in the 2 groups with naringin, although it is more

significant in the monotherapy group, these results although lower than expected, are

PT
supported by previous studies, such as published by Alam Ashraful M. and cols [11], which

RI
explained how naringin has an effect on obesity by increasing the expression of PPAR,

SC
carnitine palmitoyltransferase 1 (CPT-1), and uncoupling protein 2 (UCP-2) expression in the

liver, which could reduce adiposity in rats. Also naringin promotes gene expression and
NU
regulation of adipocytes, thereby decreasing deposition in fatty tissues [10].

Naringin significantly lowers blood glucose levels, these results are found in similar studies
MA

in rats [27-29], but contrasted in a study from Reshef N et al. [30] performed in hypertensive

patients, where there were no significant changes in glucose levels, although it is possible that
D

the results were not significant because treatment was not given completely, as naringin is
E

time-dependent [31], and naringin has shown good results in studies no longer than 5 weeks
PT

of treatment. In the study by Alam Ashraful M. et al [11] similar to the mechanism of action
CE

of naringin, which explains that naringin improves insulin signaling, for higher glucose

uptake in skeletal muscle also improves liver enzyme profile.

AC

Following the results of this study, we observed a similar reduction in the group naringin

combination with pravastatin, which lead us to ask, if positive results are minimal in the

action of naringin? Or antagonized by the action of pravastatin? so it would be interesting to

conduct a study of bioavailability and dose of the combination suggested in this study to

answer the questions above, since studies have concluded that pravastatin generates adverse

effects for the control of glucose [32].

13
 ACCEPTED MANUSCRIPT

Control obese rats showed significantly elevated serum levels of TGs, TC and LDL, but not

HDL cholesterol levels compared to the other treatment groups. The results are comparable

to those reported in a previous study [33], which supports and enables us to compare the final

results and define the effects found in the study, we can also find other studies presenting

more elevated lipid levels, for example the models induced with estreptozotocin, such as the

study by J Crespo and cols. [34], where the model shows that there are not only affects at a

PT
pancreatic level but generates higher levels of cholesterol that would help find the best

RI
treatment effects, so that new studies could be made with the combination proposed in this

SC
research but with a murine model with higher cholesterol levels.

In this study the administration of naringin or pravastatin and the combination on obese rats
NU
reduced TGs, CT and LDL compared with the control obese rats. The naringin monotherapy

group showed the better results in TGs but not in LDL and CT, the combination had the best
MA

statistical result in CT, although clinically not significant. Clinically, there were no

differences in lipids between treatment groups naringin and pravastatin and their combination,
D

it is likely that these results are presented to be given the maximum possible effect, since the
E

results of the 3 groups are similar to the levels of lipids they were in the control rats without
PT

disease. An important point to note is that the pravastatin group was not superior to naringin
CE

in any of the points studied, including the lipid profile. This study could not determine

whether the combination is better than monotherapy, so it would be interesting to conduct a

AC

similar study with a combination proposal but with higher level lipids, such as in those found

in studies in rats with invasive disease models, lipids added, to show real advantages over

monotherapy.

As for biosafety, this study shows how the combination of naringin and pravastatin is

clinically safe in the model of obese rats, these results were expected, first by the decision to

separate treatments in its administration, so we sought for no changes in absorption [35]; in

14
 ACCEPTED MANUSCRIPT

hepatic levels, pravastatin is the only statin that does not need the metabolism of family

CYP450, and naringin is credited its main antagonism to the liver by this family [35], besides

this, since there are no reports of adverse events by naringin [10], and pravastatin is usually a

safe treatment; although in the present study we found a "plus", because the use of naringin

could reduce the adverse effects in the liver by pravastatin, in the group of obese rats with

monotherapy of pravastatin observed significant elevations of hepatic transaminases, which

PT
are not observed in the group where naringin was added, so the question arisen, naringin is a

RI
liver protector against pravastatin? Does there exist a change in bioavailability? Could

SC
naringin reduce the remaining adverse effects of pravastatin? And other statins? The results of

this study opens up opportunities for new and more comprehensive studies, there is a need for
NU
more studies with this combination to show the positive effects that it could exert, if it works,

it could be an interesting tool for treating dyslipidemia by reducing adverse effects and
MA

increasing the attachment of those patients who seek alternative therapies.

D

Interest conflict. The authors declare that they have no conflict of interest
E
PT

ACKNOWLEDGEMENT

Thanks to the animal house, the University of Guadalajara for allowing the study, care and the
CE

contribution of experimental animals.

AC

FUNDING

This work was supported by the Doctorate of Pharmacology, Physiology Department, and

University of Guadalajara.

15
 ACCEPTED MANUSCRIPT

REFERENCES

1) Gutiérrez JP, Rivera-Dommarco J, Shamah-Levy T, Villalpando-Hernández S, Franco

A, Cuevas-Nasu L, Romero-Martínez M, Hernández-Ávila M. Encuesta Nacional de

Salud y Nutrición 2012. Resultados Nacionales. Cuernavaca, México: Instituto

Nacional de Salud Pública (MX), 2012.

2) WHO. Enfermedades crónicas. OMS. http://www.who.int/topics/chronic_diseases/es/

PT
3) Wacher-Rodarte N. Epidemiología del síndrome metabólico. Gac Méd Méx

RI
2009;145(5),384

SC
4) Carrillo R, Sanchez MJ, Elizondo S. Sindrome metabolico. Rev Fac Med UNAM

2006;49(03):98-104
NU
5) Han TS, Lean MEJ. A clinical perspective of obesity, metabolic syndrome and

cardiovascular disease. JRSM Cardiovasc Dis 2016;5:2048004016633371. doi:

MA

10.1177/2048004016633371

6) Gamboa-Gómez CI, Rocha-Guzmán NE, Gallegos-Infante JA, Moreno-Jiménez MR,

D

Vázquez-Cabral BD, González-Laredo RF. Plants with potential use on obesity and its
E

complications. EXCLI J 2015;14:809-831

PT

7) Patti AM, Toth PP, Giglio RV, Banach M, Noto M, Nikolic D, Montalto G, Rizzo M.
CE

Nutraceuticals as an Important Part of Combination Therapy in Dyslipidaemia. Curr

Pharm Des. 2017;23(17):2496-2503.

AC

8) Sahebkar A, Serban MC, Gluba-Brzózka A, Mikhailidis DP, Cicero AF, Rysz J,

Banach M. Lipid-modifying effects of nutraceuticals: An evidence-based approach.

Nutrition. 2016 Nov-Dec;32(11-12):1179-92.

9) Cicero AFG, Colletti A, Bajraktari G, Descamps O, Djuric DM, Ezhov M, et al. Lipid

lowering nutraceuticals in clinical practice: position paper from an International Lipid

Expert Panel. Arch Med Sci. 2017 Aug;13(5):965-1005.

16
 ACCEPTED MANUSCRIPT

10) Bharti S, Rani N, Krishnamurthy B, Arya DS. Preclínical Evidence for the

Pharmacological Actions of Naringin: A Review. Planta Med 2014; 80(06): 437-51

11) Alam MA, Subhan N, Mahbubur M, Uddin S, Reza HM, Sarker SD. Effect of Citrus

Flavonoids, Naringin and Naringenin, on Metabolic Syndrome and Their Mechanisms

of Action. Adv Nutr 2014 Jul; 5(4): 404–417. doi: 10.3945/an.113.005603

12) Mahmoud AM. Hematological alterations in diabetic rats - Role of adipocytokines and

PT
effect of citrus flavonoids. EXCLI J 2013;12: 647–657.

RI
13) Rosenson RS, Baker S, Banach M, Borow KM, Braun LT, Bruckert E, et al.

SC
Optimizing Cholesterol Treatment in Patients With Muscle Complaints. J Am Coll

Cardiol. 2017 Sep 5;70(10):1290-1301.

NU
14) Chatterjee K, Topol EJ. Fármacos en Cardiología. Cardona- Muñoz EG (editor).

Panamá. Jaypee-Highlights Medical Publishers, Inc 2014:157-72

MA

15) ATP III, Third Report of the National Cholesterol Education Program (NCEP) Expert

Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults

D

(Adult Treatment Panel III) Final Report National Cholesterol Education Program
E

National Heart, Lung, and Blood Institute National Institutes of Health. NIH 2002;
PT

Publication No. 02-5215

CE

16) Fryhofer SA. Switching From Simvastatin 80 mg: How to Shop for

Statins. Medscape 2011.

AC

17) Fukazawa I, Uchida N, Uchida E, and Yasuhara H. Effects of grapefruit juice on

pharmacokinetics of atorvastatin and pravastatin in Japanese. Br J Clin Pharmacol

2004; 57(4):448–55. doi: 10.1046/j.1365-2125.2003.02030.x

18) NIH. United States Department of Health and Human Services, Public Health Service,

National Institutes of Health. Guide for the Care and Use of Laboratory Animals; NIH

(National Institutes of Health) 1985:85-23

17
 ACCEPTED MANUSCRIPT

19) Raffoul-Orozco AK, Ávila-González AE, Barajas-Vega JL, Rodríguez-Razón CM,

García-Cobián TA, Ramírez-Lizardo EJ, Rubio Arellano ED. Impact of Cassia

acutifolia Infusion on Glucose Levels in Obesity and Diabetes Rat Model. J

Pharmacopuncture 2017;20(2):201-206

20) Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-

density lipoprotein cholesterol in plasma, without use of the preparative

PT
ultracentrifuge. Clin. Chem 1972;18:499–502.

RI
21) Erejuwa OO, Nwobodo NN, Akpan JL, Okorie UA, Ezeonu CT, Ezeokpo BC, et al.

SC
Nigerian Honey Ameliorates Hyperglycemia and Dyslipidemia in Alloxan-Induced

Diabetic Rats. Nutrients 2016;8(3).pii: E95. doi: 10.3390/nu8030095.

NU
22) Erejuwa OO, Sulaiman SA, Wahab MS, Sirajudeen KN, Salleh MS, Gurtu S.

Glibenclamide or metformin combined with honey improves glycemic control in

MA

streptozotocin-induced diabetic rats. Int. J. Biol. Sci 2011;7:244–252.

23) Garcia-Cobian TA, Rodriguez-Razon CM, Totsuka S, Cardona-Muñoz EG, Raffoul-

D

Orozco AK, García-Iglesias T. Efecto de Azadirachta Indica sobre insulina y péptido c

E

en un modelo de ratas obesas. XVII. Congreso Internacional de Avances en medicina.

PT

Guadalajara, Jalisco, Mex; 2015

CE

24) Fasanmade AA, Alabi OT. Differential effect of honey on selected variables in lloxan-

induced and fructose-induced diabetic rats. Afr. J. Biomed. Res 2008;11:191–196

AC

25) Alam MA, Kauter K, Brown L. Naringin improves diet-induced cardiovascular

dysfunction and obesity in high carbohydrate, high fat diet-fed rats. Nutrients 2013;5:

637-50

26) Li F, Zhang M, Xu D, Liu C, Zhong Z, Jia LL, et al. Co-administration of paroxetine

and pravastatin causes deregulation of glucose homeostasis in diabetic rats via

18
 ACCEPTED MANUSCRIPT

enhanced paroxetine exposure. Acta Pharmacol Sin 2014;35(6):792–805. doi:

10.1038/aps.2014.24

27) Annadurai T, Muralidharan AR, Joseph T, Hsu MJ, Thomas PA, Geraldine P.

Antihyperglycemic and antioxidant effects of a flavanone, naringenin, in

streptozotocin–nicotinamide-induced experimental diabetic rats. J Physiol Biochem

2012;68:307–318

PT
28) Annadurai T, Thomas PA, Geraldine P. Ameliorative effect of naringenin on

RI
hyperglycemia-mediated inflammation in hepatic and pancreatic tissues of Wistar rats

SC
with streptozotocin- nicotinamide-induced experimental diabetes mellitus. Free Radic

Res 2013;47:793–803
NU
29) Tsai S-J, Huang C-S, Mong M-C, Kam W-Y, Huang H-Y, Yin M-C. Anti-

inflammatory and antifibrotic effects of naringenin in diabetic mice. J Agric Food

MA

Chem 2012;60:514–521

30) Reshef N, Hayari Y, Goren C, Boaz M, Madar Z, Knobler H. Antihypertensive Effect

D

of Sweetie Fruit in Patients with Stage I Hypertension. Am J Hypertens

E

2005;18(10):1360-3
PT

31) So-Yeon K, Hye-Jin K, Mi-Kyung L, Seon-Min J, Gyeong-Min D, Eun-Young K, et

CE

al. Naringin Time-Dependently Lowers Hepatic Cholesterol Biosynthesis and Plasma

Cholesterol in Rats Fed High-Fat and High-Cholesterol Diet. J Med Food

AC

2006;9(4):582–586

32) Lorza-Gil E, Salerno AG, Wanschel AC, Vettorazzi JF, Ferreira MS, Rentz T, et al.

Chronic use of pravastatin reduces insulin exocytosis and increases β-cell death in

hypercholesterolemic mice. Toxicology. 2016 Feb 17;344-346:42-52. doi:

10.1016/j.tox.2015.12.007.

19
 ACCEPTED MANUSCRIPT

33) Bailey DG, Arnold JM, Strong HA, Munoz C, Spence JD. Effect of grapefruit juice

and naringin on nisoldipine pharmacokinetics. Clin Pharmacol Ther 1993;54(6):589-

94.

34) Crespo MJ, Quidgley J. Simvastatin, atorvastatin, and pravastatin equally improve the

hemodynamic status of diabetic rats. World J Diabetes 2015;6(10):1168–1178. doi:

10.4239/wjd.v6.i10.1168

PT
35) Koitabashi Y, Kumai T, Matsumoto N, Watanabe M, Sekine S, Yanagida Y, et al.

RI
Orange juice increased the bioavailability of pravastatin, 3-hydroxy-3-methylglutaryl

SC
CoA reductase inhibitor, in rats and healthy human subjects. Life Sci

2006;78(24):2852-2859
NU
MA
E D
PT
CE
AC

20