Anti-Inflammatory Activity of Abutilon indicum Linn. Leaf

 

DK Golwala1*, LD Patel2, SB Bothara1, SK Vaidya 1, AR Sahu1 and S Kumar3

1C.U. Shah College of Pharmacy and Research, Wadhwan, Gujarat, India.               

2 SAL Institute of Pharmacy, Sola-Bhadaj Road, Ahmedabad, Gujarat, India.          

3 GHB College of Pharmacy, Anayad, Panchmahal, Gujarat, India.

*Corresponding Author E-mail: golwala@hotmail.com

 

ABSTRACT:

The present study reveal that the ethanolic and aqueous extracts of Abutilon indicum Leaf using two different doses of each extract in acute (Carrageenin induced hind paw edema) and subacute (Formaldehyde induced hind paw edema) inflammation induced in rats, p.o. administration. In acute inflammation the % protection AIE 300 mg/kg - 64.04 % and 100 mg/kg - 59.55%, while AIA 300 mg/kg - 46.66 % and 100 mg/kg - 32.88 %. In Subacute inflammation the % protection in 3rd h are AIE 300 mg/kg - 86 % and 100 mg/kg - 68 %, while AIA 300 mg/kg - 68 % and 100 mg/kg - 44 %.The % protection of both the extracts shows Abutilon indicum possess potent anti-inflammatory activity and it significantly reduces inflammation.

 

KEYWORDS: Abutilon indicum, Anti-inflammatory, carrageenin and formaldehyde-induced edema. 

 


 

INTRODUCTION:

Abutilon indicum species is commonly called as “Country mallow” in English, commonly known as “Thuthi” is distributed throughout the hotter parts of India. Abutilon indicum is known as Atibala in Sanskrit. It has been widely used as medicine in Ayurvedic system of medicine. Abutilon indicum (Malvaceae)1.

 

Phytoconstituents like β-Sitosterol (0.2 %)2,3, tocopherol oil (0.3%)3 were isolated. Abutilon indicum is rich of fatty acids like linoleic acid, oleic acid, palmitic acid, stearic acid and capric acid. Along with vanillin, p-coumaric acid, p-hydroxybenzoic acid, caffeic acid and fumaric acid. p-β-D-glucosycoxybenozoic acid, gluco-vanilloyl glucose, fructose, galactase, glucose, leucine, histidine, threonine, serine, glutamic acid and aspartic acid2,4. Two sesquiterpene lactones Alantolactone, isoalanto-lactone5. Gallic acid6, it also contains flavonoids like luteolin, chrysoseriol, luteolin-7-O-β gluco pyranoside, chrysoseriol 7-O-β gluco pyranoside, apigenin 7-O-β-gluco pyranoside, Quercetin 3-O-β-gluco pyranoside, Quercetin 3-O-α rhamnopyranosyl, β-gluco pyranoside7. The phytochemical studies on Abutilon indicum revealed the presence of steroids, sapogenins, carbohydrates, coumarins and flavonoids8.

 

Leaf decoction is used as eyewash, mouthwash in toothache and tender gums; gonorrhea and inflammation of the bladder.

 

The whole plant is used for anti-inflammatory, immuno stimulating effect and in piles. Flower paste is applied to boils and ulcers. Seeds are considered laxative in piles, aphrodisiac, in treatment of urinary disorders, gonorrhea and chronic cystitis9. It has been reputed in the siddha system of medicine as a remedy for jaundice, piles, ulcer and leprosy10.

 

In some places, juice from the leaves of the plant is used in combination with the liquid extract of Allium cepa to treat jaundice and hepatoprotective studies on experimental animals confirmed the above activity11,12. The plant is reported to have analgesic13-15, hepatoprotective16, hypoglycemic activity17, wound healing activity18, antidiabetic19, anti-diarrhoeal20, immunomodulatory21,22, antimalarial23, antifertility24, ACE inhibitory activity25, aqueous extract was found very potent to reduce blood sugar levels26 and antimicrobial activity27. Acute oral toxicity studies were reported 4000 mg/kg12,16However, there are no reports on the anti-inflammatory activity of Abutilon indicum Linn the present study is an attempt to validate it.

 

MATERIALS AND METHODS:

Plant material:

Abutilon indicum leaves were collected during the month of January 2009, from Muradabad, Uttar Pradesh, India and authenticated by Dr. P. Jayraman, Chennai. The fresh leaves were separated and kept for shade drying. Dried leaf material was powdered using mechanical grinder and passed through 60 mesh sieve to get the powder of desired coarseness. Powdered material was preserved in an air tight container.


Table 1: Effect of AIE and AIA on Carrageenin induced hind paw edema in rats.

Group

Treatment

Dose mg/kg

Carrageenin-induced paw edema volume (ml)

Percentage protection

Group I

Control

--

0.89 ± 0.12

 

Group II

ASA

300

0.22 ± 0.03***

75.28

Group III

AIE

100

0.36 ± 0.06**

59.55

Group IV

AIE

300

0.32 ± 0.05**

64.04

Group V

AIA

100

0.60 ± 0.09

32.58

Group VI

AIA

300

0.440±0.07

46.66

Values are expressed as mean ± SEM; n=6,  *** p<0.001, **p<0.01, *p<0.05 Drug/test substance Vs Control

 

Table 2: Effect of AIE and AIA on formaldehyde induced hind paw edema in rats.

Group

Treatment

Dose

(mg/

kg)

Paw edema volume

1st hour

%  REV

2nd hour

%

REV

3rdhour

%

REV

24th hour

%  REV

Group I

Control

--

0.18± 0.006

--

0.275±0.025

--

0.25± 0.016

--

0.99 ± 0.05

--

Group II

Diclofenac

sodium,

10

0.16±0.009

11.11

0.1±0.011***

63.63

0.025±0.007***

90

0.36 ± 0.04***

50.68

Group III

AIE

100

0.09±0.014**

50

0.195±0.026

29.09

0.08±0.006***

68

0.45 ± 0.02***

38.36

Group IV

AIE

300

0.035±0.004***

80.55

0.075±0.007***

72.72

0.035±0.007***

86

0.38 ± 0.03***

47.95

Group V

AIA

100

0.175±0.030

2.77

0.205±0.020

25.45

0.14±0.02

44

0.85 ± 0.03

14.14

Group VI

AIA

300

0.1±0.013*

44.44

0.105±0.012***

61.81

0.08±0.011***

68

0.53 ± 0.03**

27.39

%REV: percentage reduction edema volume. Values are expressed as mean ± SEM; n=6. *** p<0.001, **p<0.01, *p<0.05. Drug/extracts substance Vs control.

 

 


Preparation of extracts:

Dried Abutilon indicum leaf powder was subjected to successive extraction in a soxhlet extractor using ethanol and water28. The extracts were filtered and the filtrates were concentrated under reduced pressure to obtain the extracts as solid residues29.

 

Preliminary Phytochemical studies:

Preliminary phytochemical test of Abutilon indicum ethanolic and aqueous extracts were performed for phytochemical analysis of alkaloids, glycosides, carbohydrates, steroids, tannins, phenolic compounds, proteins, amino acids, saponins, flavonoids, terpenoids, gums and mucilages29,30.

 

Animals:

Male Wistar rats weighing 150-200 g of either sex were procured from Cadila Health Care Ltd., Dholka, Ahmedabad. All the animals were kept in standard polypropylene cages and maintained under standard conditions: temperature (24 ± 10 C), relative humidity (45-55 %) and 12:12 light: dark cycle. The animals were fed with standard rat pallet and water ad libitum. The animals were allowed to acclimatize to laboratory conditions 48 h before the start of the experiment. A group of 6 rats (150-200 gm) were taken and were treated as follows in both the models. Animals in Group I served as control were, treated with vehicle (water) orally. Group II received standard drug as per different models. Group III and Group IV received AIE at the dose levels of 100 mg/kg and 300 mg/kg p.o., respectively. Group V and Group VI received AIA at the dose levels of 100 mg/kg and 300 mg/kg p.o., respectively. All the experiments were conducted after obtaining permission from the Institutional Animal Ethics Committee (IAEC).

Anti-inflammatory activity evaluation:

Carrageenin-induced hind paw edema in rats:

The acute hind paw edema was produced by injecting 0.1 ml of Carrageenin (prepared as 1% suspension in 1% CMC) locally into the plantar aponeurosis of the right hind paw of rats31. AIE and AIA (100 mg/kg and 300 mg/kg p.o.) was administered to four different groups while Group I served as control received vehicle (1 ml/kg, p.o.).Group II received standard drug, acetylsalicylic acid (ASA, 300 mg/kg, p.o.). All the groups were administered there respective doses 1 h prior to the injection of carrageenin. The rat pedal volume up to the ankle joint was measured using plethysmometer at 0 h (just before) and 3 h after the injection of carrageenin. Increase in the paw edema volume was considered as the difference between 0 and 3 h. Percent inhibition of edema volume between treated and control groups was calculated as follows:

 

                Percent inhibition =  Vc Vt    × 100

                                                      Vc

Where, Vo = Volume of the paw of control at time‘t’. Vt = Volume of the paw of drug treated at time‘t’. Where Vc and Vt represent the mean increase in paw volume in control and treated groups, respectively.

 

Formalin induced paw edema:

Male or female Wistar rats with a body weight between 150-200 g are used. The animals are starved overnight. To ensure uniform hydration, the rats receive 5 ml of water by stomach tube (Group-I). The different groups of rats were administered with AIE and AIA (100 mg/kg and 300 mg/kg p.o.) dissolved or suspended in the same volume of water. Diclofenac sodium standard drug (10 mg/kg p.o.) was given to Group-II32.  Thirty minutes later; the all the groups were administered with an injection of 0.1 ml of 1% formalin into the plantar region of the left hind paw. The paw was marked with ink at the level of the lateral malleoulus and immersed in mercury up to this mark. The paw volume was measured plethysmographically immediately after drug administration and then at 1, 2, 3 and 24 h after formaldehyde injection. For comparison purpose, the volume of oedema at various prefixed time intervals was measured. The difference between paw volumes of the treated animals was measured and the mean oedema volume was calculated33. Compare the mean percent change in paw volume. Percentage reduction in oedema volume was calculated by using the formula,

 

                Percentage reduction =   Vo - Vt   x 100

                                                              Vo

Where, Vo = Volume of the paw of control at time‘t’. Vt = Volume of the paw of drug treated at time‘t’. From the data obtained, the mean oedema volume and percentage reduction in oedema was calculated.

 

Statistical Analysis:

The results were subjected to statistical analysis by using one- way ANOVA followed by Tukey- Kramer test to calculate the significance difference if any among the groups. P<0.05 was considered as significant.

 

RESULTS:

Preliminary Phytochemical studies:

The preliminary phytochemical screening of ethanolic and aqueous extract shows the presence of alkaloids, glycosides, carbohydrates, tannins, phenolic compounds, proteins, amino acids, saponins, flavonoids, terpenoids, gums and mucilages.

 

Carrageenin-induced hind paw edema in rats:

The mean increase in paw edema volume was about 0.89±0.12 ml in the vehicle-treated control rats. AIE (100 and 300 mg/kg, p.o.) significantly (p < 0.01) reduced the mean paw edema volume at 3 h after carrageenin injection. AIE (100 and 300 mg/kg, p.o.) exhibited anti-inflammatory activity in a dose-dependent manner with the percent inhibition of paw edema of 59.55 and 64.04 respectively, as compared with the control group. While AIA (100 and 300 mg/kg, p.o.) showed 32.58 and 46.66 respectively, percent inhibition of paw edema as compared with control. However, the standard drug, ASA (300 mg/kg, p.o.) showed highly significant (p < 0.001) anti-inflammatory activity with the percent inhibition of 75.28 % (Table 1).

 

Formalin induced paw edema:

AIE (100 and 300 mg/kg, p.o.) significantly diminished the mean paw edema volume at 1 h (50 % and 80.55 %, p<0.01 and p < 0.001 respectively); 2 h AIE 300 mg/kg p.o. (72.72 % p < 0.001), 3 h (68 % and 86 % respectively, p < 0.001) and 24 h (38.36% and 47.95% respectively, p< 0.001). AIA with a dose of 300 mg/kg showed significant activity during the experimental time period. The maximum inhibition of edema volume produced by AIE (300 mg/kg, p.o.) was almost comparable to that of Diclofenac Sodium (10 mg/kg, p.o.) (86 % versus 90 % at 3 h). Interestingly, the effect of AIE persisted up to a period of 24 h in contrast to Diclofenac sodium (Table 2).

 

DISCUSSION:

The present study demonstrated anti-inflammatory activity of ethanolic and aqueous extract of Abutilon indicum in acute inflammation (Carrageenin induced hind paw edema in rats) and subacute (Formalin induced paw edema), thereby indicating the possibility of developing the extract as potent anti-inflammatory therapeutic agent. The leaf of Abutilon indicum has been used in traditional system of medicine for treating various ailments like toothache, tender gums, gonorrhea, inflammation etc. In acute and subacute models of inflammation upon challenges by phlogistic stimuli, AIE and AIA, showed significant anti-inflammatory activity.

 

The edema and inflammation induced by carrageenin is showed to be mediated by histamine and 5-HT, during 1st h. after which increased vascular permeability is maintained by the release of kinins up to 2.30 h and from 2.30 to 6 h, the mediators appear to be prostaglandins, the release of which is closely associated with migration of leucocytes into the inflamed site34. The Carrageenin induced paw edema model in rats is known to be sensitive to cycloxygenase (COX) inhibitors and has been used to evaluate the effect of non-steroidal anti-inflammatory agents35. In autacoid-induced inflammations, AIE produced significant inhibitory activity against histamine, 5-HT and PGE2 induced hind paw edema in rats.

 

Inflammation induced by formaldehyde is biphasic, an early neurogenic component is mediated by substance P and bradykinin followed by a tissue mediated response where histamine, 5-HT, prostaglandins and bradykinin are known to be involved36. In the formaldehyde-induced inflammation, the AIE and AIA demonstrated significantly anti-inflammatory activity that lasted up to 24 h in contrast to Diclofenac sodium, which was effective only at 1.5 h, suggesting its long duration of action.

 

Here we compare the both extracts with two different doses in comparision we find out the both models that alcoholic extract of Abutilon indicum 300 mg/kg dose shows significant anti-inflammatory activity while comparing with the control and other dose of aqueous extract. In conclusion, the present study clearly showed that AIE 300 mg/kg possessed good anti-inflammatory activity and also scientifically validated the traditional use of this plant for treating inflammatory disorders in the folk medicine. During the course of present work it is observed that Abutilon indicum leaf extracts possess significant pharmacological activity and hence work should be extended to study to identify the possible mechanism of action as well as establishing the therapeutic value in the treatment of inflammatory diseases.

 

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Received on 25.08.2009          Modified on 23.10.2009

Accepted on 22.11.2009         © RJPT All right reserved

Research J. Pharm. and Tech. 3(1): Jan.-Mar. 2010; Page 183-186