Anti-Hemorrhoidal Activity of Leaf Extract of Adenia lobata (Jacq.) Engl. (Passifloraceae)

 

Konkon N’dri Gilles1,2, Adjoungoua Attoli Léopold1, Ouattara Djakalia2,  Simaga Dédéou1,  Koné Bamba1, N’guessan Kouakou Edouard2 and  Kouakou Tanoh Hilaire3*

1Laboratoire de Pharmacognosie, UFR des Sciences Pharmaceutiques et Biologiques, Université Cocody

01 BP V 34 Abidjan 01, Côte d’Ivoire;

2Laboratoire de Botanique, UFR Biosciences, Université Cocody, 22 BP 1414 Abidjan 22, Côte d’Ivoire;

3 Laboratoire de Biologie et d’Amélioration des Productions Végétales, UFR Sciences de la Nature, Université

d’Abobo-Adjamé, 02 BP 801 Abidjan 02, Côte d’Ivoire.

Corresponding author: tanohilaire@yahoo.fr

 

ABSTRACT:

Next to modern therapeutic, many plants resulting from the African traditional pharmacopeia are used by the traditional healers as anti-hemorrhoidal remedies. The aim of this study was to evaluate the anti-hemorrhoidal activity of leaves of Adenia lobata. This work has focused on the inflammatory aspect of the hemorrhoid. Induction of inflammation was made with kaolin mixed with arabic gum. Solutions of A. lobata leaves used with preventive measure delay the inflammatory process of hemorrhoidal disease and for curative cure hemorrhoidal inflammation. Also, the anti-inflammatory activity observed is due to flavonoids present in leaves of A. lobata. Preparation of solutions with cool water is more effective than that carried out with the hot water which denatures the active ingredients of the remedy. At the doses used, this remedy is deprived of acute toxicity and can be taken safety by patients. A six times higher solution concentration than that of the traditional healer gives a similar anti-inflammatory response to it. Thus, there is a minimum concentration of A. lobata leaves solutions from which the anti-inflammatory effect is maximal. The anti-hemorrhoidal action of leaves of A. lobata was established and tests on veinotonic aspect are in progress.

 

KEYWORDS: Adenia lobata; anti-hemorrhoidal activity; Côte d’Ivoire; leaf; traditional healer

 


INTRODUCTION:

In Côte d’Ivoire, for economic reasons, much of the population turned to folk medicine where the traditional healers propose different herbal remedies to cure their disease. The use of the plants for therapeutic purposes is reported in the literatures1-3. Hemorrhoid extremely common disease and its prevalence is nearly 30% of the population; this disease affects one adult in two older than 50 years4. Hemorrhoids are vascular cushions, consisting of thick sub mucosa containing both venous and arterial blood vessels, smooth muscle and elastic connective tissue5-7. While, everyone has this tissue, it is the enlargement, bleeding and protrusion due to venous insufficiency that create pathology and its manifestations are an extremely common medical problem that every physician should be prepared to treat.

 

The modern drugs used in the treatment of hemorrhoids are classified in two groups of which the therapeutic effects are complementary: anti-inflammatory drugs and the veinotonic ones with vitamin-p activity8-10. Also, treatment includes over-the-counter topical medications, surgery and herbal therapy11-13. A major component of a safe and effective therapy for hemorrhoids, often overlooked, is the use of plant extracts. Several plant extracts have been shown to improve microcirculation, capillary flow and vascular tone and strengthen connective tissue of the peri vascular amorphous substrate. Early intervention with plant therapies may prevent time-consuming and expensive complications of hemorrhoids14. In effect, extract of Ruscus aculeatus is effective in increasing venous tone because of its anti-inflammatory and astringent properties. The active biochemical constituent is proposed to be the saponin glycoside ruscogenin6. Adenia lobata is used extensively in the treatment of icterus, of the cephalous, of the otitis. Of other activities of this plant are signaled mainly the anti-hemorrhoidal activity15,16. In addition, several studies showed the presence of flavonoids, sterols and terpenoids1,2,17,18 in leaves of Adenia lobata. These bioactive compounds have a variety of properties and according some reports, they possess anti-inflammatory activities10,20. A number of traditional medicine products contain plant compounds. To date, complementary medicine continues to be essential to solve people´s health problems regardless of gender, marital status, education, socioeconomic status, place of residence and religious affinity21,22.

 

Despite the impressive pharmacologic activity of phytochemicals compound of adenia lobata, the benefits that the decoctions of this plant can play in hemorrhoid therapy are not highlighted. The purpose of this study is to investigate the use of decoction adenia lobata in reducing the inflammatory response in hemorrhoidal affection. In this study, the anti-hemorrhoidal activities, in particular the inflammatory aspect of hemorrhoid, were studied by activity test on mices in which we induced in experiments the disease using the phlogogene substances such as kaolin. Lastly, various doses of the decoction were administrated in order to define an effective daily posology.

 

MATERIAL AND METHODS:

Plant material:

Leaves of Adenia lobata (Jacq.) Engl. (Passifloraceae) were taken on the grounds of the University of Cocody (Abidjan- Côte d’Ivoire) and used in this study. It is a climbing liana. Plant can spring until the top of the highest trees. It is characterized by its foliar polymorphism but especially by the aspect of its stem which is covered with galls giving appearance of “toad skin” and whose size can reach that of the thigh23,24.

 

Animals:

For this study white mice were provided by vivarium of higher teacher training school and then kept in animalery of the Training and Research Unity of Pharmaceutical and Biological Sciences at University of Abidjan-Cocody. They are 10 weeks old. The mice were fed with FACI (Fabrication d’Aliments de Côte d'Ivoire) pellets, groundnuts and dried fish. Their drink was tap water.

 

Preparation of solutions:

Five (5) different solutions were used in this study:

1- Solution A: crude extract from leaves of Adenia lobata. This solution contains a mixture of flavonoids, sterols and polyterpenes at 26.0 mg/ml 2,17,25,26. It was prepared with cold distilled water;

2- Solution B: contains flavonoids extract from leaves of Adenia lobata at 26.0 mg/ml according to methods of plant flavonoids extraction27-29. It was prepared with hot distilled water;

3- Solution C: contains crude extract from leaves of Adenia lobata. This solution contains a mixture of flavonoids, sterols and polyterpenes at 4.30 mg/ml after phytochemistry analysis2,17,25,26. It was prepared with hot distilled water (formulation of traditional healer);

4- Solution D: contains flavonoids extract from leaves of Adenia lobata at 20.0 mg/ml according to methods of plant flavonoids extraction27-29. It was prepared with cold distilled water;

5- Solution E: kaolin suspension. It was obtained by dissolving 10 g of kaolin powder in a gum arabic solution at 5g/100 ml (in boiling water). The resulting suspension was injected around the edge of mice anus using a 1 ml syringe fitted with a needle according to the technique of Adjimi30 and Koné31.

 

Methods:

The demonstration of anti-inflammatory activity was performed using the method of Winter32. Briefly, it consists in injecting carrageenin under the plantar fascia of the hind leg of mice. It then produces an inflammatory reaction which is characterized by the appearance of an oedema. The intensity of the oedema is evaluated by the increase in paw volume expressed as a percentage compared to initial volume. Preventive administration by intraperitoneally way of an anti-inflammatory drug significantly reduced the development of oedema. In this study, induction of the inflammatory response was made by injecting a gum arabic and kaolin suspension (phlogogene substances that are cheap) at the rim of mice anus. The administration of the various solutions was then performed as follows:

 

1- Experiment 1: inflammatory induction of hemorrhoidal disease by injection of kaolin suspension:

Two (2) groups of 4 mices were used:

-group 1 or control group: injection of water distilled in various points around the anus of mices;
-group 2: injection of kaolin suspension on the edge of the anus of mices.

 

2- Experiment 2: anti-hemorrhoidal activity of solutions of Adenia lobata leaves:

Four (4) groups of 4 mices were used:

- Group 1: no treatment after inflammatory induction;

- Group 2: solutions were administered one hour before the induction inflammation at a dose of 0.20 ml/20 g of mice body weight;

- Group 3: solutions were administered 2 times daily at 0.20 ml/20 g of mice body weight of the 1st day of the inflammatory signs appearance until the 5th day.

- Group 4: solutions were administered 2 times daily at 0.20 ml/20 g of mice body weight at the onset inflammation signs until the 5th day.

 

At the end, a visual and magnifying glass appreciation of inflammatory regression until the total disappearance of the signs (mice completely cured) was conducted for 7 days. Similarly, changes can support over time such as inflammatory signs duration and latency time i.e. period between the injection time of kaolin suspension of the kaolin and appear it signs of the inflammation were raised.


RESULTS:

1- Inflammatory induction of hemorrhoidal disease by injection of kaolin suspension

Distilled water has caused no changes around the anus of mices. On the other hand, with the injection of the kaolin suspension, an inflammation was observed. The anus of the mices presents redness, prolapse and gaping as well as mucus (Table 1).

Table 1. Response of the mice after injection of the kaolin suspension

Group of mices

Observations

1

Healthy anus without change

 

2

Presence of inflammation :

- redness

-prolapse
-gaping
- mucus

 

2. Anti-hemorrhoidal activity of solutions of Adenia lobata leaves

Without treatment (group 1: control), the inflammation is induced one hour after the injection of the kaolin solution and lasts 7 days. All solutions administered to mices one hour before the induction of inflammation (group 2) had a latency time higher than 2 h. Solution A have the highest latency (2.25 h = 2h15 min) followed by solutions C and B. However, the solution D has the lowest latency time (2.10 h = 2h06 min). Signs of inflammation lasted 6 days in mices treated with solutions A, B and C. On the other hand, in mices treated with solution D, signs of inflammation lasted only 5 days (Table 2).

 

Table 2. Latency time and duration of inflammation in mice of group 2

 

Latence time (h)

Duration of

inflammation (day)

Solution A

2.25

6

Solution B

2.16

6

Solution C

2.23

6

Solution D

2.10

5

 

The results of the effect of the solutions administered to mices after the appearance inflammation signs are show in Table 3. Solutions A and C gave the most important latency times followed by the solutions D and C. Level of group 3, the duration of the inflammation was 4 days in mices treated with solutions A, C and D and 5 days in mices treated with solution B. Moreover, the duration of inflammation was 5 days in mices treated with solutions A, B and C while it is 3 days in mices treated with solution D.

 

Table 3. Latency time and duration of inflammation in mice of groups 3 and 4

 

Latence time (h)

D.I.  (day)

Group 3

Group 4

Solution A

2.43

4

5

Solution B

1.72

5

5

Solution C

2.5

4

5

Solution D

1.88

4

3

DI: duration of inflammation

 

The evaluation of inflammation regression until no signs (completely healed mices) was carried out during 7 days. Thus, on a set of 4 mices (group 4), the healing is from the 3rd day and is complete in all mices treated with solutions A and C on the 5th day. It is the same for the mices treated with solution B, but the full recovery of all mices is located on the 6th day. On the other hand, with solution D, the healing appears the 2nd day of treatment and mices are cured on the 3rd day of treatment (Table 4).

 

 

Table 4. Evaluation of healed mices during 7 days of treatment

 

Day 1

Day 2

Day3

Day 4

Day 5

Day 6

Day 7

Solution A

0

0

1

3

4

-

-

Solution B

0

0

1

2

3

4

-

Solution C

0

0

1

3

4

-

-

Solution D

0

1

4

-

-

-

-

 

DISCUSSION:
This work has focused on the inflammatory aspect of the hemorrhoid. The induction of inflammation was done with substances such as the kaolin and arabic gum well known for their phlogogene power. The inflammatory type of hemorrhoid appears with a latency time one hour on average. Adjimi30 reported a 30 min latency time with kaolin used as single substance phlogogene. On the other hand, this latency time increases to 2 h or longer when the mices were previously treated at least 30 min immediately with Adenia lobata solutions, before induction of hemorrhoidal disease. Thus, the use of preventive solutions of Adenia lobata appears to retard the inflammatory process of hemorrhoidal disease.

 

Without treatment, the inflammation was resolved in 8 days. The use of Adenia lobata solutions for curative, significantly change the duration of induced inflammation. Thus, it was noted a resorption time of 4 days for the solutions A and C; 5 days for the solution B and 3 days for solution D. It follows that these solutions are active in the inflammatory aspect of the hemorrhoidal disease. By comparing the activity of solutions A and B, we noted that the temperature of water used for the extraction plays a key role in the duration of resorption of inflammation. Indeed, analysis of Table 4 shows that the disappearance of inflammation occurs more rapidly with solution A than solution B for identical concentration of flavonoids. It should be noted that these two solutions differ one from the other by the presence of sterols and polyterpenes in solution B. Similarly, with the solution D containing only the total flavonoids, the cure was even faster. This seems to suggest that the anti-inflammatory drug activity related to the flavonoids present in leaves of Adenia lobata2,10,18,33. Indeed, according to the literature, the flavonoids (in particular rustoside and diosmin) enter in the treatment of hemorrhoidal disease. They work by increasing venous tonicity and capillary resistance, while reducing capillary permeability33-35. Flavonoids present in the plant leaves could have a mechanism action similar to those existing or new molecules to be identified which would be necessary for the marketing of new drugs. In the same way, sterols and polyterpenes would have a secondary anti-inflammatory activity20. Indeed, their presence in solution A would involve a synergy of action. They could also have a veinotonic action because of share their constitution, sterols and polyterpenes have a chemical structure near of that to the vitamin P, well-known for its veinotonic properties9,10. However, in the drug treatment of hemorrhoidal disease, veinotonic and anti-inflammatory action are complementary20; this could explain that solution A acts faster than solution B, which contains only flavonoids. It would be interesting to isolate these sterols and polyterpenes to study their anti-inflammatory and veinotonic activities in order to confirm the hypothesis. The administration of solutions A and C with different concentrations (26.0 mg/ml solution A and 4.30 mg/ml the solution C), rectal enema form, gives a similar anti-inflammatory response. Thus, for different concentrations, the anti-inflammatory activity observed is the same. In this study, a concentration 6 times higher than that of the traditional healer (solution C) gives an identical anti-inflammatory response to the latter. This would be due to the existence of an active minimal concentration from which anti-inflammatory effect is maximal19,36. The determination of this concentration will prevent the waste of Adenia lobata leaves but also to treat several patients with a small amount of leaves. In addition, for the same route of administration, responses observed are identical when the concentrations of leaves solutions are different. For this study, the rectal preparations which are administered as enema, there is an almost total rejection of solutions in a short time. Thus, in fact only the time is passed between the administration and the rejection that allows a contact solutions with the areas affected. This brief contact, repeated several times, regardless of the solution concentration, ends up inducing anti-inflammatory response. However, the enema did allow a sufficient contact time; it is understandable that the activity of solutions A and C is identical. Any extension of the contact time would promote sufficient impregnation of the areas affected by the solutions and thus optimize the discounted effect. It would be interesting to develop an other formulations such as ointments or the suppositories that would maintain constant contact and prolonged with the anal lesions37. This would increase the anti-inflammatory activity of Adenia lobata and significantly reduce the resorption time of inflammation.

 

ACKNOWLEDGEMENTS:

We thank Professor Aké-Assi, Director of the Floristic National Center of Côte d’Ivoire, for the botanical identification of Adenia lobata.

 

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Received on 24.08.2011          Modified on 04.09.2011

Accepted on 11.09.2011         © RJPT All right reserved

Research J. Pharm. and Tech. 5(1): Jan. 2012; Page 63-67