In vivo Evaluation of Aloe vera and Ficus deltoidea Anti-inflammatory effect in Mice: A Brief Update

 

Nurul A. Rozkhaidi1, Elysha N. Ismail2, Reezal Ishak1*

1Institute of Medical Science Technology, Universiti Kuala Lumpur, A1, 1, Jalan TKS 1,

Taman Kajang Sentral, 43000 Kajang, Selangor, Malaysia.

2Department of Biomedical Science, Faculty of Medicine and Health Sciences,

Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.

*Corresponding Author E-mail: reezal@unikl.edu.my

 

ABSTRACT:

Aloe vera (Family: Liliaceae) and Ficus deltoidea (Family: Moraceae), also known as Mas Cotek has been applied in traditional medicines as sources of anti-inflammatory remedy and as therapeutic alternatives to conventional medicines. The present study was carried out to evaluate the in vivo anti-inflammatory effect of Aloe vera and Ficus deltoidea powder extracts in mice. The mice were divided into 4 groups (n=5), namely the negative control, positive control and two treatment group of Aloe vera and Ficus deltoidea extracts. The animals were treated with normal saline, diclofenac, Aloe vera and Ficus deltoidea extracts according to their respective group, 30 minutes before carrageenan was introduced. The inflammation was induced by injecting carrageenan into the sub-plantar region of left hind paw of all mice. The paw edema was measured using digital caliper at 0, 0.5, 1, 2, 3, 24 and 48 hours after carrageenan was injected. There were significant differences in the measurement of the paw edema between normal saline (negative control), diclofenac (positive control), Aloe vera and Ficus deltoidea powder extract at p<0.05. Diclofenac (10mg/kg) significantly inhibited paw edema from 0.5 hour onwards (p<0.05); Aloe vera extracts (4000mg/kg) from the 1st hour onwards; and Ficus deltoidea extracts (4000mg/kg) from the 2nd hour onwards. The anti-inflammatory action demonstrated by both Aloe vera and Ficus deltoidea extracts are comparable to those of diclofenac treatment even though their efficacy is lesser. The findings indicate that Aloe vera and Ficus deltoidea powder extracts possess anti-inflammatory effect on carrageenan induced paw edema in vivo. Therefore, in conclusion, the Aloe vera and Ficus deltoidea powder extract have the potential in managing acute inflammation and serve as an alternative therapeutic agent.

 

KEYWORDS:  Aloe vera, Ficus deltoidea, Carrageenan, Anti-inflammatory, Mice, Paw edema.

 

 

 INTRODUCTION: 

Inflammation involves a localize complex defensive reaction of tissues in response to injury, infection, irritation, hypersensitivity, or toxicity. It is characterized by the presence of swelling, redness, heat, pain, and may cause a loss of function. Many evidence shows a close association between inflammation and chronic health conditions that suggests inflammation may lead to the pathophysiological condition of chronic diseases1. In general, the inflammatory response shares a common mechanism of action although the processes depend on the exact nature of the initial stimulus and its location in the body. The mechanism involves the recognition of harmful stimuli by the cell surface pattern receptor which triggers the activation of inflammatory pathway before inflammatory markers are released for the recruitment of inflammatory cells to the site2. The chemical mediators released during the inflammatory reaction include vasoactive amines such as serotonin and histamine, peptide in the form of bradykinin, and eicosanoids namely prostaglandins, leukotrienes, and thromboxane3. The current commercially available medication to treat acute and short term inflammation are usually local or systemic anti-inflammatory drugs such as corticosteroids and NSAIDs. These medications have various drawbacks, hence justify the need for better, safe, potent, non-toxic, or less toxic inflammatory drugs. Medicinal plants have been the central subject for treating various diseases nowadays and their isolated natural derivatives compounds are used in conventional medicine for the treatment of various inflammatory disorders4,5.

 

Aloe species or scientifically identified as Aloe vera or Aloe barbadensis have been used for thousands of years as traditional herbal medicine around the world6,7. Aloe vera bulk as well as its extracts are often used in cosmetic product, skincare, healthcare, food and beverages, and in medical industry as active ingredients for extra hygiene, therapeutic and rejuvenating purpose8. Both the gel and latex from the leaves of Aloe vera are used for different application. The clear gel from the plant is usually rubbed onto the skin as an ointment for treating wounds and burns, whereas the leaf’s green portion can be turned into juice, or dried and used as oral laxative9. Additionally, in many commercial products various forms of Aloe vera is added in beverages, capsules, distillates, powders and as flavouring10. Aloe vera is also best known for improving skin textures and prevent wrinkles11. Aloe vera contains several physiologically active compounds that shows anti-inflammatory effects, immunomodulatory, and wound healing properties. The bioactive compounds include polysaccharides, glycoproteins, anthraquinones, and low molecular weight biological components that might act alone or in concert12.

 

Ficus deltoidea, commonly known as Mistletoe Fig, or Mas Cotek in Malaysia, is a large shrub or small tree species native to the Southeast Asia and belongs to Moraceae family. The leaf of Ficus deltoidea measured from 3 to 5cm and width from 3 to 4cm, is deltoid in shape with a distinctive forked midrib. The leaf is light green and leathery with rusty golden spots on the upper side and yellowish green with black spots on the lower side, while the base of the leaf is cuneate and apex shape which is rounded or truncate13. Ficus deltoidea is studied for its diverse organic properties with the modern pharmacological studies showing that this plant has a wide range of beneficial attributes for human health. The use of this plant as an alternative medicinal herb is getting diverse recognition where Ficus deltoidea tea bags and capsules are commercially sold in the local market mainly in the Southeast Asia region. It is believed that the leaves of Ficus deltoidea contains high amount of magnesium, manganese, and potassium, hence tea made from this leaves are deemed as good source of daily minerals for human consumption14. Furthermore, the fruit from different varieties of Ficus deltoidea also showed effective antidiabetic properties15, 16. The pharmacological properties reported from the extracts of Ficus deltoidea includes anti-diabetic, antioxidant, anti-inflammation, wound healing, and anti-cancer17,18.

 

MATERIALS AND METHODS:

Materials:

The Aloe vera and Ficus deltoidea powder extracts were kind donation from the Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Malaysia and produced using spray drying process. The Aloe vera and Ficus deltoidea powder extracts were dissolved in normal saline to a dose of 4000mg/kg. The standard drug, diclofenac, was prepared freshly each time and is administered intraperitoneally. Diclofenac was administered in a volume of 10ml/kg/dose19.

 

Laboratory animal model:

White male Balb/c mice, weighing between 20 to 30g, were used in this study. The mice were housed in polypropylene cages in groups of five mice per cage, and were kept under controlled environment condition with free access to water and food ad libitium20,21. The ethical clearance for the animal study was obtained from the Animal Ethics Committee (AEC), Institute of Medical Science Technology Universiti Kuala Lumpur (FYP/AEC/MESTECH-UNIKL/2020/July).

 

Experimental design:

For the purpose of the study, the animals were divided into five groups with five animals in each group.

Group I (Control): Received normal saline 0.05ml through intraperitoneal injection + carrageenan (0.05 ml, sub-plantar).

Group II (Standard): Received standard drug diclofenac (10mg/kg) through intraperitoneal injection+ carrageenan (0.05ml, sub-plantar).

Group III (Treatment): Received Aloe vera treatment (4000mg/kg) through intraperitoneal injection + carrageenan (0.05ml, sub-plantar).

Group IV (Treatment): Received Ficus deltoidea treatment (4000mg/kg) through intraperitoneal injection + carrageenan (0.05ml, sub-plantar).

 

The chemical compound used to induce inflammatory reaction in the animals is carrageenan.

 

Carrageenan induced paw edema in mice:

Inflammation on the paw of the animals were induced by injecting carrageenan (0.1ml, 1%w/v in physiological saline) into the sub-plantar region of the left hind paw of each mouse22. Prior to the procedure, Aloe vera and Ficus deltoidea extracts were administered intraperitoneally to the animals in Group III and IV respectively, 30 minutes before carrageenan was injected. Normal saline and diclofenac were administered to the control and standard group respectively.

 

The thicknesses of the paw were measured using digital calliper. The initial paw thicknesses were measured at 0 hour. After that, the paw thicknesses were measured at 0.5, 1, 2, 3, 24 and 48hours after the carrageenan injection and the results were recorded and tabulated accordingly. The percentage of inhibition of edema was calculated using the following formula23:

 

                                           T - T0    

% Inhibition of edema = --------------- X 100

                                              T0

T: Thickness of paw in control group

T0: Thickness of paw edema in the treated groups.

 

Statistical analysis:

The value for edema measurement were analyzed using One-Way ANOVA followed by Dunnett’s test and are expressed as mean±standard deviation (SD). Differences between the mean value of the treated animals and control groups were considered statistically significant at p<0.05.

 

RESULT AND DISCUSSION:

The present study was designed to evaluate the anti-inflammatory effect of Aloe vera and Ficus deltoidea extracts in mice model. Carrageenan induced acute inflammation is one of the widely applied procedure and most suitable to evaluate the effectiveness of potential anti-inflammatory agents.

 

Based on Table 1, a gradual increase in the measurement of the mice paw edema was recorded by Group I (Normal saline) after carrageenan injection, and it reached the maximum size on the 3rd hour (3.08mm) as compared to the initial thickness in the beginning (1.66 mm). The effect of Aloe vera and Ficus deltoidea extracts on carrageenan induced inflammation in mice at different hours of study was compared to the measurement shown by Group I for the evaluation of anti-inflammatory reaction on the basis of percentage inhibition of paw edema. The time course of edema development after carrageenan injection onto the paw of the animals illustrates a biphasic effect. The first phase of inflammation was observed within two hours after the injection. It is believed that the increase vascular permeability in the first hour is implicated to the release of histamine and serotonin, whereas the second phase and onwards is related to the presence of kinins, and subsequently the release of prostaglandins, lysosomes, and protease24,25.

 

Based on figure 1, the presence of diclofenac in the physiological system appears to inhibit the development of edema significantly from 0.5 hour onwards. Aloe vera administered to the mice at the dose of 4000mg/kg did not indicate any significant inhibition on 0.5 hour, but it showed significant inhibition (p<0.05) from the 1st hour until 48 hours. Whereas, Ficus deltoidea did not demonstrate any inhibitory effects from 0.5 to 1 hour, but produced significant inhibition (p<0.05) from the 2nd hours onwards. Based on these findings, it can be inferred that the inflammation inhibitory effects of Aloe vera and Ficus deltoidea powder extract may be due to the inhibition of primary mediators such as histamine and serotonin in the first phase, and as a result, the cyclooxygenase pathways were inhibited in the second phase of the anti-inflammatory effect26,27.

 

 

Table 1: Effect of Aloe vera and Ficus deltoidea powder extract on carrageenan induced paw edema in mice.

Treatment Group (n=5)

Dose

0 hr

(mm)

0.5 hr

(mm)

1 hr

(mm)

2 hr

(mm)

3 hr

(mm)

24 hr

(mm)

48 hr

(mm)

Group I

Normal saline

1.66 ± 0.04

2.93 ± 0.11

2.96 ± 0.11

3.01 ± 0.10

3.08 ± 0.11

2.75 ± 0.17

2.25 ± 0.09

Group II

Diclofenac (10 mg/kg)

1.65 ± 0.04

2.39 ± 0.29

2.11± 0.23

2.00 ± 0.16

2.34 ± 0.24

1.84 ± 0.10

1.70 ± 0.04

Group III

Aloe vera

(4000 mg/kg)

1.63 ± 0.01

2.89 ± 0.10

2.72 ± 0.11

2.40 ± 0.11

2.27 ± 0.10

1.97 ± 0.16

1.84 ± 0.16

Ficus deltoidea

4000 mg/kg

1.69 ± 0.03

2.12 ± 0.03

1.97 ± 0.04

2.66 ± 0.06

2.44 ± 0.04

2.12 ± 0.04

1.97 ± 0.04

 

 

 

 

 

 

 

 

 

One way

ANOVA

F

2.43

12.68

33.96

71.58

32.13

49.66

30.10

df

3

3

3

3

3

3

3

p value

0.1

<0.001

<0.001

<0.001

<0.001

<0.001

<0.001

Values are expressed as mean of paw edema thickness ± standard deviation (SD). Time intervals were measured from the time of carrageenan injection. There is significant difference between normal saline (Group I), diclofenac (Group II), Aloe vera (Group III) and Ficus deltoidea (Group IV) powder extract at p<0.05. Intergroup comparison was performed using Dunnett multiple comparisons test. p<0.05 compared with normal saline (Group I) in the same duration.

 

The findings from this study are supported by previous report on Aloe vera inhibitory actions on the cyclooxygenase pathway and reduction of prostaglandin E2 production from arachidonic acid28,29. The phenolic compounds found in Ficus deltoidea, namely flavonoids, have also been reported to obstruct the protein kinase C, and L-arginine or nitric oxide (NO) pathways, before consequently halt the production of prostaglandin E­2 (PGE2) by suppressing the inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2)30.

 

Figure 1: Percentage of paw edema inhibition after treatment with diclofenac, Aloe vera and Ficus deltoidea powder extract on carrageenan induced paw edema at different hours. *p<0.05 compared with normal saline group (Group I).

 

Comparison between Aloe vera and Ficus deltoidea treatment to the diclofenac treated group proved that the control drug is still more effective than both Aloe vera and Ficus deltoidea powder extracts dosage of 4000 mg/kg in reducing the induced paw edema. However, the mice treated with both Aloe vera and Ficus deltoidea exhibited comparable anti-inflammatory effects to the treatment with diclofenac until the 48th hour period of the study. This suggests that the extracts administered to the animals are endowed with potent anti-inflammatory properties in vivo. β-sitosterol, campesterol, lupeol and cholesterol are found in aloe sterol, where these phytochemical constituents are known anti-inflammatory compounds in nature which can help in relieving inflammatory pain and function as natural analgesic28. The presence of flavonoids, polyphenols, and tannins in Ficus deltoidea has been previously reported31, and it is believed that several mechanism of actions could be responsible for the anti-inflammatory activities of Ficus deltoidea based on the existing phytochemical constituents26,30.

 

CONCLUSION:

The present study showed that Aloe vera and Ficus deltoidea powder extract possess anti-inflammatory properties against carrageenan induced paw edema in the white Balb/c mice. Carrageenan induced paw edema has been established as one of the inflammation model in order to assess the activity of natural product associated with pathological changes during inflammatory reaction, and the results of this study is an indication that Aloe vera and Ficus deltoidea powder extract are effective in combating acute inflammatory disorders. In addition, this study highlights the potential of inexpensive and effective alternatives as anti-inflammatory agents coming from natural resources, and with further research works could unravel the exact mechanism of actions involved in the anti-inflammatory activities of Aloe vera and Ficus deltoidea.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

 

ACKNOWLEDGEMENTS:

The authors gratefully acknowledge Universiti Kuala Lumpur through its FYP course for the financial support to complete this study, and the Institute of Bioproduct Development, Universiti Teknologi Malaysia (UTM), Malaysia for their kind materials donation.

 

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Received on 25.04.2024      Revised on 24.08.2024

Accepted on 01.11.2024      Published on 28.01.2025

Available online from February 27, 2025

Research J. Pharmacy and Technology. 2025;18(2):852-856.

DOI: 10.52711/0974-360X.2025.00125

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