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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 9  |  Issue : 4  |  Page : 509-516

Assessment of anti-inflammatory activity of Tridax procumbens in wistar rats


1 Department of Pharmacology, Dev KumariRajaram Pharmacy ShikshaSansthan, Sitapur, India
2 Department of Pharmacology, Hind Institute of Medical Sciences, Sitapur, India
3 Department of Pharmacology, Goel Institute of Pharmacy and Sciences, Lucknow, India
4 Department of Pharmacology, Pt. Nagina College of Pharmacy, Azamgarh, Uttar Pradesh, India

Date of Submission09-Apr-2022
Date of Acceptance06-Dec-2022
Date of Web Publication29-Dec-2022

Correspondence Address:
Dr. Rajesh Kumar Suman
Department of Pharmacology, Hind Institute of Medical Sciences, Ataria, Sitapur 261303, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/mgmj.mgmj_45_22

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  Abstract 

Background: Tridax procumbens Linn. is a spreading herb found throughout India. Tridax procumbens is known for several potential therapeutic effects like antiviral, antioxidant, anti-infective, wound healing, insecticidal and anti-inflammatory, hepatoprotective, and antidiabetic. Novel therapies for inflammation are essential to overcome the adverse effects of existing anti-inflammatory drugs. Thus, this study was designed to evaluate anti-inflammatory activity in the experimental animal. Objective: To evaluate anti-inflammatory efficacy in experimental animalMaterials and Methods: Tridax procumbens plant leaves were taken for the study. The Plant extraction was prepared by using the Soxhlet apparatus. The ethanolic extract was used as a test drug. Indomethacin and Dexamethasone were used as control drugs. Wistar rats weighing 150g to -200g were used in the study. Results: At dosages of 200 and 400 mg kg-1, the ethanolic leaf extracts of Tridax Procumbens demonstrated substantial anti-inflammatory activity against Irish Moss-triggered inflammation. After 3 hours, 400 mgkg-1 showed a significant reduction in inflammation (48 percent), with the impact increasing after 3 hours (52 percent). The anti-inflammatory efficacy of Tridax Procumbens ethanol (EtOH) extracts was substantial and comparable to that of indomethacin (10 mg/kg). Tridax procumbens ‘s Ethanol (EtOH) leaf extract at a dose of 200 and 400 mgkg-1, demonstrated considerable anti-inflammatory action. The average weight of granulomatous surrounding tissue in the threading was considerably smaller in the Tridax procumbens extraction group after 7 days than in the comparison group. The 400 mg kg-1 dose was shown to be the most effective of the two. the greatest reduction in the production of granuloma tissue. Tridax procumbens at doses of 200 mg/kg and 400 mg/kg resulted in substantial reductions in granuloma weights of 38.16 ± 0.04 (7.4 percent inhibition) and 34.58 ± 0.04 (16.1 percent inhibition), respectively. The lower dose of 400 mg/kg resulted in a little lesser reduction in granumola weight than the conventional medication dexamethasone (28.92 ± 0.04). (29.8 percent inhibition). Conclusion: The present study concludes that Tridax Procumbens possess anti-inflammatory effects in animal models.

Keywords: Herbs, pharmacological activities, Tridax procumbens linn


How to cite this article:
Prakash S, Suman RK, Chandra G, Yadav P, Kumar M. Assessment of anti-inflammatory activity of Tridax procumbens in wistar rats. MGM J Med Sci 2022;9:509-16

How to cite this URL:
Prakash S, Suman RK, Chandra G, Yadav P, Kumar M. Assessment of anti-inflammatory activity of Tridax procumbens in wistar rats. MGM J Med Sci [serial online] 2022 [cited 2023 Feb 6];9:509-16. Available from: http://www.mgmjms.com/text.asp?2022/9/4/509/365979




  Introduction Top


Tridax procumbens extracts were reported to possess anti-inflammatory, analgesic, wound healing activity, and antidiabetic effects in animal models.[1] Tridax procumbens, commonly referred to as coat buttons, is best known as a widespread weed and pest plant.

Herbs make up a large portion of the most important forms of medicine. Herbs have been utilized for medicinal purposes since the dawn of mankind. In historical documents such as the Rigveda, the Bible, and the Quran, a few native drugs utilized in medical technology have been recorded. Over 6,000 years ago, the Chinese used flora and wildlife as medicine. Herbal medicine has been used in India for over a thousand years. Hippocrates, also known as the “Father of Medicine,” was the first to scientifically explain ailments. Indian medical systems include aromatherapy, Ayurveda, Homoeopathy, Tibetan medicine, and Naturopathy. Herbal treatment is a safe and effective technique to treat a variety of internal illnesses that most medical systems consider intractable and incurable. Its goal is to both prevent and relieve pain.[2]

Inflammation, also known as phlogosis, is the dysfunctional reaction of live tissue to wounds which results in the aggregation of blood cells and plasmatic fluid in a localized area. Many diseases can be caused, maintained, or aggravated, even though protection can be induced, maintained, or aggravated. It’s a complicated phenomenon involving biochemical and immunological influences. Inflammation can be identified by the following signs and symptoms: Calor(heat), dolor(pain), Functiolaesa(Lossoffunctions), Rubor(redness), and Tumor(Swelling) Three main forms of inflammation are known, the inflammation reaction happens in different phases, obviously regulated by separate pathways, depending on the host’s defense capacity and the length of the response.[3]

Various pharmacological activities of Tridax procumbens were reported in the last few years but its anti-inflammatory effects have not been confirmed in any scientific paper. The report presented in the study is an attempt to prove the pharmacological evaluation of the anti-inflammatory activity of Tridax procumbens leaves in rats. Novel therapies for inflammation are essential to overcome the adverse effects of existing anti-inflammatory drugs.[4] Thus, the present study has been designed to evaluate the anti-inflammatory activity of Tridax procumbens in experimental rats.


  Materials and methods Top


Plant collection and authentication

Plant collection

At the beginning of March 2021, Tridax procumbens plant leaves were harvested and eroded.

Authentication

The plant was identified Department of Pharmacognosy, Goel Institute of Pharmacy and Sciences, Lucknow.

Drying and pulverization of the plant material

The leaflets were cleansed to eliminate particulate matter and permitted to air dry in the sun for full dryness after collection and authentication. The leaf extract was then ground in a mixer without any moisture.

Plant extracts preparation

The fine powder was packed securely in the Soxhlet apparatus and recovered with solvent ethanol for 72 hrs., with intermittent shaking at 60°C. Evaporation condensed the extraction to a fraction of its initial volume Phytochemical research was conducted on the ethanolic extract of Tridax procumbens that resulted.

DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay

Blois first designed the DPPH radical scavenging test, which was used to check the antioxidant activity of the compounds. In a brief, 0.1 mM DPPH was produced in 95 percent ethanol. At a concentration of 100g/mL, this solution (1mL) was added to 3 mL of sample (methanolic leaf extract and copper nanoparticles) in ethanol. The mix was thoroughly stirred and placed away at room temperature for 30 minutes. The absorption was then determined at 517 nm with Spectrophotometry. The test was done three times using sodium bicarbonate as a reference standards reagent.[5]

The reaction mixture’s decreased absorbance indicated more free radicals scavenge ability. The percent DPPH scavenge efficiency was estimated using the equation below:

DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) scavenging effect (%) = A0 - A 1 / A0 × 100

Where A0 is the absorption of the control reactions and A1 is the absorption when the test or standard sample is present.[6]

HPTLC (High-Performance thin layer Chromatography) and TLC (Thin layer chromatography) analysis

The moving phase was 2:20.5:0.7:1 (volume by volume) methanol: formic acid: water: ethyl acetate: The sample solution (5.0 L con. 1 mgmL-1) was implemented to the plate as 9.9 mm band, using a specific computerized spray on bands syringe with a hundred and the preceding setting: length of band 10 mm, implementation rate 10 sec/L, range between 3 mm, length from the plate side. Utilizing the WIN CATS application.

Screening of phytochemicals

The ethanolic extract of Tridax procumbens was tested qualitatively for alkaloids, polysaccharides, glycosides, phenolic compounds, peptides, free amino acids, and triterpenes, among other elements.

Testforcarbohydrate

A minimal amount of extraction was mixed in 5 ml of water before being processed.

Molischtest

A few drops of - naphthol were added to the filter (20 percent in ethanol). Then 1 milliliter, of pure H2SO4 was injected with the sidewalls of the slanted test tubes, and the interaction was inspected for the creation of a violet ring.[7]

Glycosides and anthraquinones analysis - bontrager’stest

The hydroxylate was isolated with benzene after a little quantity of ethanolic extract was hydrolyzed with HCL for a few hrs. in boiling water. The benzene layer was exposed to a diluted ammonia solution and the production of red-pink color was noticed.

Legal test

The extraction was diluted in pyridine and turned basic with some drop of 10 percent sodium hydroxide before adding newly produced sodium nitroprusside and watching for blue color development.

Test for flavonoids ammonia test

Membrane filter strips were immersed in a weak aqueous extract, ammoniated, and the change in color from white -to yellow was noted.

Tests for phenolic and tannins compounds

The extractions were divided into 3 parts and diluted in distilled water One portion received 10% sodium chloride, another received 1% gelatin, and the third received gelatin salt reagent. The existence of tannins was indicated by precipitate with the former or both gelatin salt reagents. A high false test is shown by precipitate with a salt solution. The introduction of several drops of diluted 1 percent FeCl3 to extracted sample, that is resulted in a bluish or greenish-black coloring, verified positive tests.

Test for proteins and amino acids

In purified water, a tiny quantity of samples was transferred and processed.

  • (a) Biuret’stest


Add 3 drops of 0.002 percent copper sulphate to the ammoniated alkaline filter and look for a red or violet color.

  • (b) Millon’stest


6–7 droplets of Millon reagents (1 g Hg + 9 ml of fuming nitric acid solutions) were added to 2 ml of filtrate, and crimson precipitates were detected.

  • (c) Ninhydrin test


To concentrate tannins, a CH3COOPb solution was applied to the filtrate and then filtration occurs. The filtrate was placed on a chromatoy sheet, coated with ninhydrin reagents boiled at 100oC for 5 min., and the red / violet color was seen.

  • (d) Xanthoprotein test


Several drops of strong nitric acid were added to the filtrate and yellow color was detected on the side of the test tubes.

Tests for triterpenes and sterols

Till the saponification was complete, the extract was vortexed with alcoholic KOH. After that, the mixture was dissolved in d.H2O and diethyl ether was used to extract it. The unsaponifiable substance was submitted to the following tests after the ethereal extraction was evaporated.

Liebermann-burchard’stest

The ether-soluble remnant mixed in CHCl3 and several droplets of acetic anhydride were applied, accompanied by several droplets of strong sulphuric acid from the test tube’s sidewalls, to see if a blue to blue-red color formed.

Salkowski’sreaction

2 mL strong H2SO[4] was added to the ether soluble residual, and the creation of a yellowish ring at the connection, which develops red after two min., was seen.

Ethical approval

The study was conducted after receiving approval from the commission for control and supervising of animals (CPCSEA) and the institutional animal ethics committee (IAEC). The IAEC approval number was 2014/Po/Re/S/18 CPCSEA.

Animals

This research employed Wistar rats weighing 150g to 200g. The animals came from animals housed at Goal College Pharmacy Lucknow, India. The animals were randomly assigned to patients treated and housed in polypropylene enclosures with paddy husks as bedding when they arrived. The animals were kept at a temp of 24°C with a relative humidity of 30 - 70 percent. The A12:12 light: day cycles were used. Every animal was given free access to the water and was fed commercial pelleted chow. All experimental techniques and methods adopted in this work were approved by Goal College of Pharmacy’s Institutional Animal Ethics Committee.

Acute toxicity studies

Toxicological testing’s accepted by Organisation for Economic Co-operation and Development (OECD) guidelines. Toxicity tests were performed on the Wistar rats that were chosen. The creatures were split into 4 groups, each with three animals. Before the acute experimental method, the animals were fed overnight. Orally, rats were given extract in doses of 300 and 400 mg.kg-1 body weight. The animals were watched constantly for the very first four hours after treatment for changes in behavior such as ataxia, hyperactivity, convulsions, salivation, tremor, diarrhea, tiredness, and sleep. They also were monitored for 15 days after receiving the medicine to see if there was any mortality. To test anti-inflammatory activities in rats, 1/10 and 1/5 of the maximum tolerable dose (300 and 400 mg.kg-1, of body weight) of ethanolic leaf extracts of Tridax procumbens were used.[8]


  ANTI-INFLAMMATORY ACTIVITY Top


Irish moss-induced paw edema in rats

The rat (150g - 200g) were put into 4 subgroups (n = 6) for this study. Group I was given normal water and diet. whereas Group II was given Indomethacin (10 mg.kg-1). The ethanolic extracts of Tridaxprocumbenswere given vocally to Groups III and IV at doses of 200 mg/kg and 400 mg.kg-1, respectively.

  • G1st: Normal Control (NC)


  • G2nd:Indomethacin(10 mg/kg)


  • G3rd: Test Drug I (Ethanolic extract leaf Tridaxprocumbens 200 mg)


  • G4th:TestDrugII(EthanolicextractleafTridaxprocumbens 400 mg)


The paw width was measured at 1h, 2h,3h,and4h after Irish Moss vaccination by using digital Vernier calipers Acute inflammation was induced in the mice by administering 0.1 ml of 1 percent (w/v) Irish Moss solution into t sub planter area of the right hind paw. The animals were given the medication 1 hour before the Irish Moss was given to them.[9] Digital vernier callipers were used to assess paw thickness at 1, 2, 3, and 4 hours after the Irish Moss administration.

Cotton pellets induced granuloma method in rats

Each cotton pellet weighed 5 milligrams and was sterilized. The pellets were injected subcutaneously injection via a skin cut on the backs of the animal while they were undergoing ether anesthesia. Group II received Dexamethasone (1 mg.kg-1), while Groups III and IV received 200 mg.kg-1 and 400 mg.kg-1 of ethanolic extracts of Tridax procumbens, respectively.

  • G1st: Normal Control (NC)


  • G2nd:Dexamethasone(1 mg/kg)


  • G3rd: Test Drug I (Ethanol leaves extracts Tridaxprocumbens)


  • G4th:TestDrugII(Ethanolic leaves extractsTridaxprocumbens)


This test medication was given once a day for seven days. Diethyl ether was used to sacrifice the rat on the eighth day. The granulomas were taken out and quantified.


  Results Top


The present study is evaluating the anti-inflammatory activity of the Tridax Procumbens plant which is tested in the rat. The inflammation produced in Rat is shown in Figure. [Figure 1]. Extraction of plant compound was carried out using the Soxhlet apparatus for extraction as shown in the figure. [Figure 2]
Figure 1: Shows: Inflammation n rat paw

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Figure 2: Soxhlet apparatus for extraction

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The phytochemical analysis of the test plant was carried out. The yield analysis of plant extract shows the presence of Hexane, Ethanol, and water in 32%, 45.6%, and 15.6% respectively. [Table 1]
Table 1: Physiochemical analysis of the extract

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DPPH (2,2-Diphenyl-1-Picrylhydrazyl) analysis for antioxidants shows the presence of Hexane, Water, and ethanol-specific absorbance which is present in Table. [Table 2]
Table 2: Antioxidant analysis of extracts

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The preliminary phytochemical evaluation of the ethanolic extract of Tridax procumbens shows Alkaloid, tannin, phenolic compounds, sterol, saponin, proteins, amino acids, and a large amount of flavonoid are found in this plant. [Table 3]
Table 3: Phytochemical screening of tridax procumbens

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Anti-inflammatory activity

Inflammatory diseases are globally identified as the major cause of morbidity across the population. Treatment of chronic inflammatory diseases like rheumatoid arthritis and inflammatory bowel diseases is still a challenge due to the lack of safe and effective drugs. Finding a safe and effective drug to control inflammation has been a challenge and therefore, many animal models have been developed for the evaluation of drugs having anti-inflammatory properties.

Rats with irish moss-triggered paw edema

The anti-inflammatory activity of Tridax procumbens ethanolic leaf extracts on Irish Moss-triggered hind paw edema. At dosages of 200 and 400 mgkg-1, the ethanolic leaf extracts of Tridax Procumbens had substantial anti-inflammatory activity against Irish Moss-triggered inflammation. After 3 hours, the concentration of 400 mgkg-1 showed a reduction of 48 percent, with the impact increasing after 3 hours (52 percent). The anti-inflammatory efficacy of Tridax Procumbens. Ethanolic extracts were substantial and comparable to that of indomethacin (10 mg/kg). [Table 4]
Table 4: Anti-inflammatory activity of ethanolic (EtOH) extracts of Tridax procumbens on Irish Moss triggered paw edema method in Wistar rats

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The anti-inflammatory activity of Tridax procumbens ethanolic leaf extracts was studied in Wistar rats utilizing the cotton pellet-generated granuloma method. Tridax procumbens ethanolic leaf extract at a dose of 200 and 400 mgkg-1, demonstrated considerable anti-inflammatory action. The average weight of granulomatous surrounding tissue in the threading was considerably smaller in the Tridax procumbens extraction group after 7 days than in the comparison group. The 400-mgkg-1 dose was shown to be the most effective of the two. the greatest reduction in the production of granuloma tissue. Tridax Procumbens at doses of 200 mg/kg and 400 mg/kg resulted in substantial reductions in granuloma weights of 38.16 ± 0.04 (7.4 percent inhibition) and 34.58 ± 0.04 (16.1 percent inhibition), respectively. The lower dose of 400 mg/kg resulted in a little lesser reduction in granumola weight than the usual medication dexamethasone (28.92 ± 0.04). (29.8 percent inhibition). [Figure 3] and [Figure 4]
Figure 3: Tridaxprocumbensethanolic leaf extract has anti-inflammatory action in Wistar rats using the Irish Moss generated paw edoema technique

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Figure 4: Tridaxprocumbensethanolic leaf extract has anti-inflammatory action in Wistar rats using the Irish Moss triggered paw edoema technique. The results are given as a percentage of inhibition

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Granuloma treatment in rats triggered by cotton pellets

The anti-inflammatory activity of Tridax procumbens ethanolic leaf extracts was studied in Wistar rats utilizing the cotton pellet-generated granuloma method. Tridax Procumbens ‘s ethanolic leaf extract at a dose of 200 and 400 mgkg-1, demonstrated considerable anti-inflammatory action. The average weight of granulomatous surrounding tissue in the threading was considerably smaller in the Tridax Procumbens extraction group after 7 days than in the comparison group. The 400-mgkg-1 dose was shown to be the most effective of the two. the greatest reduction in the production of granuloma tissue. Tridax procumbens at doses of 200 mg/kg and 400 mg/kg resulted in substantial reductions in granuloma weights of 38.16 ± 0.04 (7.4 percent inhibition) and 34.58 ± 0.04 (16.1 percent inhibition), respectively. The lower dose of 400 mg/kg resulted in a little lesser reduction in granumola weight than the usual medication dexamethasone (28.92 ± 0.04). (29.8 percent inhibition). [Table 5] [Figure 5] and [Figure 6]
Table 5: Anti-inflammation efficacy of an ethanolic extract of Tridax procumbens cotton pellets in Wistar rats with granuloma pouches

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Figure 5: Tridaxprocumbensethanolic leaf extracts have anti-inflammatory effects on cotton pellet-induced granuloma in rats

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Figure 6: On cotton pellet-induced granuloma in rats, Ethanolic leaves extract of Tridaxprocumbens has an anti-inflammatory effect. The percentage inhibition is used to express the results

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  Discussion Top


The ethanolic extract of Tridax procumbens exhibited anti-inflammatory action in a rat paw edema experimental model caused by Irish Moss. Leukocyte movement to wounded tissues is widely established to be an important element of the inflammation response. The initial inflammatory reaction is mediated by histamine and serotonin, but the protracted reaction is mediated by kinins and prostaglandins. Several plants’ anti-inflammatory properties have been linked to their significant sterol/triterpene or flavonoid contents. In mice with Irish Moss-triggered paw inflammation, ethanolic extract of Tridax procumbens demonstrated significant anti-inflammatory activity.[10] The inflammatory granuloma is a common reaction to a prolonged inflammation condition and the mass of the pellets is closely connected to the granulomatous tissue.

The present study demonstrates the anti-inflammatory activity of Tridax procumbent in rat paw edema Irish Moss and cotton pellet-produced granuloma experimental models. Prabhu et al[4] also studied the anti-inflammatory and analgesic activity of Tridax procumbent and similar results as per our present study were reported. Prolonged inflammation is caused by the proliferation of proliferative cells. These cells might be granuloma-like or stretched out. In cotton pellet-produced granuloma, the Tridax procumbens extract demonstrated potent anti-inflammatory action and may be useful in a chronic severe inflammatory state. It demonstrated its ability to reduce the amount of fibroblast and the production of collagen and mucopolysaccharide throughout the formation of granuloma tissue.


  Conclusion Top


The ethanolic leaf extract of Tridax procumbens has been shown to possess significant anti-inflammatory effects in rats.

Ethical consideration

Ethical approval was taken from the commission for control and supervising of animals (CPCSEA) and the institutional animal ethics committee (IAEC), application number IAEC the inspection is carried out (2014/Po/Re/S/18 CPCSEA).

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Kakkar M, Behl T, Cruz CV, Makeen HA, Albratty M, Alhazmi HA, et al. Tridax procumbens ameliorates streptozotocin-induced diabetic neuropathy in rats via modulating angiogenic, inflammatory, and oxidative pathways. Evid Based Complement Alternat Med 2022;2022:1795405.  Back to cited text no. 1
    
2.
Petrovska BB Historical review of medicinal plants’ usage. Pharmacogn Rev 2012;6:1-5.  Back to cited text no. 2
    
3.
Madri JA Inflammation and Healing. In: Kissane JM, editor. Anderson’s Pathology. 9th ed. St. Louis: C.V. Mosby; 1990. p. 67-10.  Back to cited text no. 3
    
4.
Prabhu VV, Nalini G, Chidambaranathan N, Kisan SS Evaluation of anti inflammatory and analgesic activity of tridax procumbens linn against formalin, acetic acid and cfa induced pain models. Int J Pharm Pharm Sci 2011;3:126-30.  Back to cited text no. 4
    
5.
Patel JK, Patel PY Botanical therapeutics: Discovery, development and manufacture-prospects and constraints. J Nat Remedies 2007:7:19-30.  Back to cited text no. 5
    
6.
Sharma P Investigation on photodecomposition of standardised ethyl acetate fraction of katha. Pharmacogn. J 2020;12:815-20  Back to cited text no. 6
    
7.
Khandelwal KR Textbook of Practical Pharmacognosy. 5thed. Pune: Nirali Prakashan; 2004. p. 149-55.  Back to cited text no. 7
    
8.
Saini NK, Singhal M Anti-inflammatory, analgesic and antipyretic activity of methanolic tecomaria capensis leaves extract. Asian Pac J Trop Biomed 2012;2:870-4.  Back to cited text no. 8
    
9.
Okokon JE, Nwafor PA Antiinflammatory, analgesic and antipyretic activities of ethanolic root extract of croton zambesicus. Pak J Pharm Sci 2010;23:385-92.  Back to cited text no. 9
    
10.
Perez RM, Perez S, Zavala MA, Salazar M Anti-inflammatory activity of the bark of hippocratea excelsa. J Ethnopharmacol 1995;47:85-90.  Back to cited text no. 10
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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