Large scale screening of commonly used Iranian traditional medicinal plants against urease activity

  • Farzaneh Nabati1,

    Affiliated with

    • Faraz Mojab2,

      Affiliated with

      • Mehran Habibi-Rezaei3,

        Affiliated with

        • Kowsar Bagherzadeh1,

          Affiliated with

          • Massoud Amanlou1, 4 and

            Affiliated with

            • Behnam Yousefi5Email author

              Affiliated with

              DARU Journal of Pharmaceutical Sciences201220:72

              DOI: 10.1186/2008-2231-20-72

              Received: 17 July 2012

              Accepted: 17 July 2012

              Published: 31 October 2012

              Abstract

              Background and purpose of the study

              H. pylori infection is an important etiologic impetus usually leading to gastric disease and urease enzyme is the most crucial role is to protect the bacteria in the acidic environment of the stomach. Then urease inhibitors would increase sensitivity of the bacteria in acidic medium.

              Methods

              137 Iranian traditional medicinal plants were examined against Jack bean urease activity by Berthelot reaction. Each herb was extracted using 50% aqueous methanol. The more effective extracts were further tested and their IC50 values were determined.

              Results

              37 plants out of the 137 crude extracts revealed strong urease inhibitory activity (more than 70% inhibition against urease activity at 10 mg/ml concentration). Nine of the whole studied plants crude extracts were found as the most effective with IC50 values less than 500 μg/ml including; Rheum ribes, Sambucus ebulus, Pistachia lentiscus, Myrtus communis, Areca catechu, Citrus aurantifolia, Myristica fragrans, Cinnamomum zeylanicum and Nicotiana tabacum.

              Conclusions

              The most potent urease inhibitory was observed for Sambucus ebulus and Rheum ribes extracts with IC50 values of 57 and 92 μg/ml, respectively.

              Keywords

              Urease inhibitor Iranian traditional medicinal plants Sambucus ebulus Rheum ribes Screening of natural products

              Introduction

              Ureases (urea amidohydrolases, EC (3.5.1.5) are a group of widespread enzymes in nature, classified as the most proficient enzymes (with proficiency more than 1014), stand as protagonist in biochemistry for several reasons. Urease was the first ureolytic enzyme obtained and named in the late nineteenth century, with landmark significance in enzymology as the first enzyme crystallized (in 1926 by Sumner) to approve the proteinous nature of the enzymes [1]. Also, as ascertained by Dixon et al. in 1975, urease was the first enzyme shown to possess nickel ions in its active site, essential for activity [2]. Since its substrate; urea is pervasively available in nature, urease was important to provide organisms with nitrogen in the form of ammonia for growth [3]. Despite the diversity in the molecular structures of urease, the amino acid sequences of the active sites are principally similar in all of the known them and consequence of this fact is the same catalytic mechanism. The active sites are always located in α subunits and contain the binuclear nickel centre, in which the Ni–Ni distances range from 3.5 to 3.7 Angstrom [4].

              Urease as the most characteristic feature of Helicobacter pylori constitutes 5–10% of the bacteria’s proteins. H. pylori a microaerophilic, gram-negative spiral bacterium which was first detected in 1984 by Marshall et al, is one of the most common chronic bacterial pathogens in humans [5]. Approximately more than 50% of people in the world are infected with it, and its prevalence is significantly higher in developing countries in compare with the developed ones. H. pylori infection is an important etiologic impetus usually leading to chronic gastritis, gastro duodenal ulcer and low grade gastric mucosa-associated lymphoid tissue lymphoma. Epidemiological data show that high H. pylori infection rate, result in the incidence of gastric cancer and adenocarcinoma [6, 7]. Urease catalyzes the hydrolysis of urea to produce ammonia and carbon dioxide, and the most crucial role is to protect the bacteria in the acidic environment of the stomach [8]. It has been also reported that ammonia and monochloramine, which is a reaction product of ammonia and hypochlorous acid, exhibit potent toxicity in gastric epithelium [9]. Moreover, it has been demonstrated that H. pylori lacking urease activity are incapable of causing infection in animal models. Thus, it is most likely that urease is essential for bacterial colonization and perhaps the pathogenesis of related disease in vivo.

              World Health Organization (WHO) has categorized H. pylori as a class 1 carcinogen [10]. Fortunately, its eradication with antibiotics can result in ulcer healing, prevent peptic ulcer recurrence and reduce the prevalence of gastric cancer in high-risk populations. However, it is not always successful because of its resistance to one or more antibiotics and other factors such as poor patient compliance, undesirable side effects of the drugs and significant cost of combination therapy [11]. Wolle et. al. reported that approximately 20% of the patients undergoing antibiotics therapy would experience therapeutic failure [12]. In developing countries, since the application of antibiotics is still under a poor management as a whole, there is a growing need for finding new anti-H. pylori agents that can hopefully eradicate the invasion and presence of survived H. pylori strains to avoid relapse of gastric ulcer. Hence, a considerable variety of studies involving tests for medicinal plants showing antimicrobial activity and discrepant susceptibility test results are available due to variations in the methods and conditions used for its susceptibility testing.

              One of the best sources of new substances to treat H. pylori is natural products and their derivatives [13]. Variety of techniques such as synthesizing [6], and also molecular modeling and virtual screening methods [14, 15] have been applied to find possible urease inhibitors. The biological activity of plant-derived substances may be considered as a source of new anti-H. pylori drugs come from different classes of compounds and are characterized by the diversity of their structures. Therefore, almost all traditional Iranian herbal medicines that are used as remedies and sold as medicines to manage different diseases were screened to discover possible plant-derived urease inhibitors.

              Methods

              Materials

              Sodium nitroprusside (sodium pentacyanonitrosyloferrate III) and urease (EC 3.5.1.5) from Jack beans were purchased from Sigma (St. Louis, MO, USA). All other chemicals were of analytical reagent grade from Merck. Deionized water was used in all experiments. Potassium phosphate buffer (100 mM), pH 7.6 was prepared in distilled water.

              The studied plants were collected from local medicinal herb shops, Tehran, Iran (June 2010) and were identified by one of our authors of the presented article (F. Mojab). The authenticated samples were deposited in the Herbarium of Shahid Beheshti University of Medical Sciences.

              Extract preparation

              10 g of air-dried and powdered plant material was extracted in 10 ml, 50:50 methanol: water at room temperature for 24 hrs. The resulting liquid extract was filtered and concentrated to dryness under reduced pressure. The dry extracts were stored at -20°C till used [16].

              Determination of urease activity

              All extracts were tested for urease inhibitory at concentration of 10 mg/ml by the modified spectrophotometric method developed by Berthelot reaction [17]. For herbal extracts that were proven to exert significant inhibition and also for positive controls, inhibitory assays were performed. The plant extracts were tested in a concentration range of 0 to 10 mg/ml. Hydroxyurea was used as standard inhibitor.

              The solution assay mixture consisted of urea (30 mM) and (100 μl) crud extract with a total value of 950 μl. The reactions were initiated by the addition of 50 μl of urease enzyme solution in phosphate buffer (100 mM, pH 7.6, 1 mg/ml). Urease activity was determined by measuring ammonia concentration after 15 minutes of enzymatic reaction. The ammonia was determined using 500 μl of solution A (contained 5.0 g phenol and 25 mg of sodium nitroprusside in 500 ml of distilled water) and 500 μl of solution B (contained of 2.5 g sodium hydroxide and 4.2 ml of sodium hypochlorite 5% in 500 ml of distillated water) at 37°C for 30 minutes. The absorbance was read at 625 nm. Activity of uninhibited urease was designated as the control activity of 100%.

              Data processing

              The extent of the enzymatic reaction was calculated based on the following equation:
              I % = 1 T / C * 100 http://static-content.springer.com/image/art%3A10.1186%2F2008-2231-20-72/MediaObjects/40199_2012_Article_72_Equa_HTML.gif

              Where I (%) is the inhibition of the enzyme, T (test) is the absorbance of the tested sample (plant extract or positive control in the solvent) in the presence of enzyme, C (control) is the absorbance of the solvent in the presence of enzyme. Data are expressed as mean ± standard error (SD) and the results were taken from at least three times.

              Determination of IC50 values

              IC50 values (concentration of test compounds that inhibits the hydrolysis of substrates by 50%) were determined by studying the extracts urease inhibitory activity at their different concentrations in comparison to their individual positive control employing spectrophotometric measurement. IC50 values were obtained from dose-response curves by linear regression, using Graphpad software, prism 5.

              Results and discussion

              Medicinal plants as an appropriate and renewable source of active chemical compounds can be used as templates to discover new lead compounds. Doxorubicin, vincristine, and taxol, are examples of these herbal compounds which are clinically applied. According to the literature, 50% of commercially presented medicines in 1985 was from herbal origins [18]. Gastrointestinal diseases, especially gastric, duodenal and peptic ulcer, arise from different factors, in particular microbial agent H. pylori. Common multi-drug therapies not only have side effects, but are also expensive. On the other hand, the probability of drug resistance occurrence and disease retrogression is quite concerning. Already reported studies have shown that herbal compounds have the ability to prevail this microbe. Among the studied herbal essences and extracts, many did not exceed the study level due to production limits, toxicity and impossibility of drug form preparation. Majority of the researches have focused on ways to inhibit the bacteria growth or it’s elimination from the culture, while a few has particularly concerned inhibition of urease enzyme which is responsible for the bacteria defense system against the stomach very acidic medium.

              Specific inhibition or reduction of urease enzyme activity would result in an increased sensitivity of the bacteria in acidic medium and therefore it’s natural elimination by stomach acidic condition or the body immune system.

              In the presented study, urease enzyme inhibition potency of 137 herbal extracts was investigated from which 37 extracts have shown inhibitory activity up to more than 70% in the concentration of 10 mg/ml (Table 1). Further examinations and IC50 determination revealed that Sambucus ebulus, Rheum ribes, Pistachia lentiscus, Myrtus communis, Myristica fragrans, Areca catechu, Cinnamomum zeylanicum, Citrus aurantifolia and Nicotiana tabacum extracts inhibit urease enzyme in concentrations less than 500 μg/ml. It should also be mentioned that C. zeylanicum, M. chamomilla and M. spicata are already used as gastrointestinal remedies and this research has proved that these herbs can inhibit urease activity and prevent gastric upsets. Names of the studied plants and the 37 final more active extracts are presented in Table 2. As it is shown, the most potent urease inhibitory was observed for S. ebulus and R. ribes with IC50 values less than 100 μg/ml.
              Table 1

              Urease inhibitory activity of plants extract at concentration of 10 mg/ml

               

              Scientific name

              Plant family

              Common name in English

              Common name in Persian

              Part used in traditional

              Inhibition (%)

              1.

              Abrus precatorius

              Fabaceae

              Paternoster Seed

              Cheshm-e khorus

              Seed

              9.21 ± 0.04

              2.

              Acacia Senegal

              Fabaceae

              Gum Arabic

              Samgh-e arabi

              Gum

              12.81± 0.09

              3.

              Acanthophyllum squarrosum

              Asparagaceae

              Soap Root

              Chubak

              Root

              14.15 ± 0.02

              4.

              Alpinia officinarum

              Zingiberaceae

              Galangal

              Khulanjan

              Rhizome

              41.75 ± 0.05

              5.

              Althaea officinalis

              Malvaceae

              Hollyhoch

              Khatmi

              Flower

              20.94 ± 0.06

              6.

              Alyssum homolocarpum

              Brassicaceae

              Madword & Pepper Weed

              Qodume

              Seed

              13.57± 0.12

              7.

              Amaranthus lividus

              Amaranthaceae

              Cock's Comb Seed

              Tokhm-e tajkhorus

              Seed

              17.48 ± 0.11

              8.

              Anethum graveolins

              Apiaceae

              Dill Seed

              Tokhm-e shevid

              Seed

              37.50 ± 0.03

              9.

              Apium graveolens

              Apiaceae

              Celery Seed

              Tokhm-e karafs

              Seed

              2.43 ± 0.01

              10.

              Aquilaria sinensis

              Thymelaeaceae

              Agarwood

              Udeqamari

              Fruit

              32.03 ± 0.08

              11.

              Arctium Lappa

              Asteraceae

              Burdock Root

              Bâbââdam

              Root

              19.99 ± 0.08

              12.

              Areca catechu

              Arecaceae

              Betel Nuts

              Fufel

              Fruit

              96.67 ± 0.01

              13.

              Artemisia absinthium

              Asteraceae

              Worm Wood

              Afsantin

              Herb

              52.50 ± 0.06

              14.

              Artemisia dracunculus

              Asteraceae

              Tarragon

              Tarkhon

              Leaf

              57.53 ± 0.03

              15.

              Asperugo procumbens

              Boraginaceae

              German Madwort

              Bâdranjbuye

              Herb

              12.43 ± 0.02

              16.

              Astragalus arbusculinus

              Fabaceae

              Sarcocola

              Anzarut

              Gum

              17.68 ± 0.06

              17.

              Astragalus gossypinus

              Fabaceae

              Gum Tragacanth

              Katirâ

              Gum

              1.33 ± 0.02

              18.

              Bambusa vulgaris

              Poaceae

              Golden Bamboo

              Tabâshir sadaf

              Secretions

              12.81 ± 0.04

              19.

              Brassica nigra

              Brassicaceae

              Mustard

              Khardel

              Seed

              27.63 ± 0.01

              20.

              Calendula officinalis

              Asteraceae

              Marigold

              Hamishe bahar

              Flower

              0.16 ± 0.06

              21.

              Calendula sp.

              Asteraceae

              Marigold

              Hamishe bahar

              Flower

              8.21 ± 0.07

              22.

              Camellia sinensis

              Theaceae

              Green Tea

              Chây-e sabz

              Leaf

              89.40 ± 0.02

              23.

              Camellia sinensis

              Theaceae

              Green Tea

              Châyeparsefid

              Twig

              90.45 ± 0.01

              24.

              Cannabis sativa

              Cannabaceae

              Hemp Seed

              Shâhdane

              Seed

              9.71 ± 0.02

              25.

              Capsicum annuum

              Solanaceae

              Red Pepper

              Felfel-e qermez

              Fruit

              99.01 ± 0.01

              26.

              Carthamus tinctorius

              Asteraceae

              Saf

              Golrang

              Flower

              50.78 ± 0.04

              27.

              Cassia angustifolia

              Fabaceae

              Senna

              Sena

              Leaf

              3.29 ± 0.03

              28.

              Celosia cristata

              Amaranthaceae

              Cockscomb

              Gol-e halva

              Flower

              82.55 ± 0.03

              29.

              Centaurea sp.

              Asteraceae

              Centaurea

              Gol-e gandom

              Flower

              70.33 ± 0.02

              30.

              Chenopodium botrys

              Amaranthaceae

              Lamb's Quarter

              Dermane-e torki

              Herb

              15.13 ± 0.04

              31.

              Cichorium intybus

              Asteraceae

              Chicory

              Kâsni

              Herb

              40.55 ± 0.04

              32.

              Cinchona officinalis

              Rubiaceae

              Cinchona

              Gne gne

              Bark

              67.03 ± 0.02

              33.

              Cinnamomum camphora

              Lauraceae

              Camphre

              Kâfur

              Camphor

              10.14 ± 0.08

              34.

              Cinnamomum cassia

              Lauraceae

              Cassia

              Salikhe

              Bark

              91.19 ± 0.02

              35.

              Cinnamomum zeylanicum

              Lauraceae

              Cinnamon

              Darchin

              Bark

              84.22 ± 0.05

              36.

              Citrus aurantifolia

              Rutaceae

              Limu Fruit

              Limu ammâni

              Fruit

              99.02 ± 0.02

              37.

              Citrus aurantium

              Rutaceae

              Bitter Orange Peel

              Khalâl-e nârenj

              Rind

              1.43 ± 0.05

              38.

              Citrus bigardia

              Rutaceae

              Orange

              Gol-e nârenj

              Twig

              24.31 ± 0.03

              39.

              Colchicum macrophyllum

              Colchicaceae

              Colchicum Corms

              Suranjan

              Corm

              9.44 ± 0.08

              40.

              Commiphora molmol

              Burseraceae

              Myrrh

              Morr-e Makki

              Gum

              8.22 ± 0.04

              41.

              Crataegus microphylla

              Rosaceae

              Hawthorn

              Sorkhe valik

              Flower

              82.19 ± 0.03

              42.

              Curcuma zedoaria

              Zingiberaceae

              Zedoary

              Zorombad

              Seed

              4.70 ± 0.06

              43.

              Cuscuta epithymum

              Convolvulaceae

              Hellweed

              Aftimun

              Herb

              9.66 ± 0.01

              44.

              Cymbopogon

              Poaceae

              Lemongrass

              Putar

              Root

              14.02 ± 0.03

              45.

              Descureania

              Brassicaceae

              Flixweed Seed

              Khakshir

              Seed

              21.81 ± 0.01

              46.

              Diplotaenia damavendica

              Apiaceae

              Diplotaenia

              Gozal

              Seed

              12.59 ± 0.06

              47.

              Doronicum bracteatum

              Asteraceae

              Doronicum

              Darunj-e aqrabi

              Herb

              10.73 ± 0.01

              48.

              Dracaena cinnabari

              Asparagaceae

              Dragon Blood

              Khone siyavosh

              Gum

              49.49 ± 0.13

              49.

              Dracocephalum

              Lamiaceae

              Moldavian Balm

              Badrashbi

              Twig

              3.95 ± 0.01

              50.

              Echinophora platyloba

              Apiaceae

              Echinophora

              Khosharize

              Herb

              17.48 ± 0.01

              51.

              Echium amoenum

              Boraginaceae

              Ox tongue Flower

              Gol-e gâvzabân

              Flower

              31.66 ± 0.02

              52.

              Elaeagnus angustifolia

              Elaeagnaceae

              Oleaster

              Senjed

              Fruit

              4.67 ± 0.14

              53.

              Elaeagnus angustifolia

              Elaeagnaceae

              Oleaster

              Gol-e senjed

              Flower

              27.45 ± 0.01

              54.

              Elettaria cardamomum

              Zingiberaceae

              Cardamon

              Hel sabz

              Fruit

              13.16 ± 0.04

              55.

              Elletaria cardamomum

              Zingiberaceae

              Cardamon

              Hel sefid

              Fruit

              6.80 ± 0.07

              56.

              Elletaria cardamomum

              Zingiberaceae

              Cardamon

              Hel siyah

              Fruit

              5.78 ± 0.071

              57.

              Equisetum arvense

              Equisetaceae

              Horse Tail

              Dom-e asb

              Stem

              52.35 ± 0.05

              58.

              Eruca sativa

              Brassicaceae

              Rocket

              Tokhm-e mandâb

              Seed

              13.28 ± 0.05

              59.

              Eucalyptus sp.

              Myrtaceae

              Eucalyptus

              Okaliptus

              Leaf

              47.92 ± 0.01

              60.

              Euphorbia sp.

              Euphorbiaceae

              Euphorbia

              Gav kosh

              Herb

              68.94 ± 0.03

              61.

              Ferula assa-foetida

              Umbelliferae

              Assa-Foetid

              Anqoze

              Gum

              34.07 ± 0.04

              62.

              Helicteres isora

              Malvaceae

              Screw Tree Pod

              Bahmanpich

              Fruit

              8.18 ± 0.02

              63.

              Heracleum persicum

              Apiaceae

              Cow Parsnip Friut

              Golpar

              Fruit

              10.27 ± 0.02

              64.

              Hibiscus gossypifolius

              Malvaceae

              Rose Mallow

              Chay-e Makki

              Herb

              96.28 ± 0.02

              65.

              Humulus lupulus

              Cannabaceae

              Hops

              Râzak

              Twig

              54.85 ± 0.02

              66.

              Hypericum perforatum

              Hypericaceae

              St.John's Wort

              Alaf-e chay

              Herb

              97.99 ± 0.02

              67.

              Juglans regia

              Juglandaceae

              Walnut Shell

              Pust-e vasat-e gerdo

              Septum

              93.62 ± 0.01

              68.

              Juglans regia

              Juglandaceae

              Walnut Shell

              Pust-e gerdo

              Rind

              1.27 ± 0.06

              69.

              Juniperus Sabina

              Cupressaceae

              Sabine

              Abhal

              Fruit

              19.63 ± 0.01

              70.

              Lactuca sativa

              Asteraceae

              Lettuce

              Tokhm-e Kâhu

              Seed

              2.93 ± 0.04

              71.

              Lawsonia inermis

              Lythraceae

              Henna

              Hana

              Leaf

              54.00 ± 0.06

              72.

              Levisticum officinalis

              Apiaceae

              Lovage

              Anjadân romi

              Seed

              10.00 ± 0.06

              73.

              Linum usitatissimum

              Linaceae

              Lineseed

              Tokhm-e katan

              Seed

              2.71 ± 0.18

              74.

              Malabaila secacule

              Apiaceae

              Parsnip

              Dogho

              Root

              18.18 ± 0.04

              75.

              Malva sylvestris

              Malvaceae

              Common Mallow

              Gol-e panirak

              Flower

              14.15 ± 0.05

              76.

              Matricaria chamomilla

              Asteraceae

              Chamomile

              Bâbon-e shirazi

              Herb

              87.21 ± 0.01

              77.

              Melissa officinalis

              Lamiaceae

              Balm

              Barangbu

              Herb

              46.22 ± 0.05

              78.

              Mentha spicata

              Lamiaceae

              Mint

              NaAna

              Leaf

              93.89 ± 0.01

              79.

              Myristica fragrans

              Myristicaceae

              Nutmeg

              Joz-e buya

              Fruit

              78.19 ± 0.01

              80.

              Myrtus communis

              Myrtaceae

              Myrtle

              Murd

              Leaf

              72.99 ± 0.01

              81.

              Nasturdium officinalis

              Brassicaceae

              Watercress

              Boolâgoti

              Leaf

              74.00 ± 0.03

              82.

              Nerium Oleander

              Apocynaceae

              Nerium

              Gol-e kharzahre

              Flower

              84.62 ± 0.01

              83.

              Nicotiana Tabacum

              Solanaceae

              Tobacco

              Tutun

              Leaf

              52.77 ± 0.03

              84.

              Nicotiana tabacum

              Solanaceae

              Tobacco

              Tutun

              Stem

              75.26 ± 0.05

              85.

              Nymphaea alba

              Nymphaeaceae

              White Lotus

              Gol-e nilofar

              Flower

              97.86 ± 0.01

              86.

              Ocimum basilicum

              Lamiaceae

              Basil

              Reyhan-e banafsh

              Leaf

              19.61 ± 0.05

              87.

              Ocimum basilicum

              Lamiaceae

              Basil

              Reyhan-e sabz

              Leaf

              0.41 ± 0.01

              88.

              Oenothera biennis

              Onagraceae

              Evening Star

              Gol-e maghrebi

              Flower

              3.95 ± 0.04

              89.

              Olea europea

              Oleaceae

              Olive Leaf

              Barg-e zyton

              Leaf

              72.30 ± 0.01

              90.

              Orchis latifolia

              Orchidaceae

              Oriental Salp

              SaAlab-e panjei

              Root

              18.90 ± 0.02

              91.

              Orchis mascula

              Orchidaceae

              Male Orchis

              SaAlab-e qolvei

              Root

              3.16 ± 0.04

              92.

              Papaver Rhoeas

              Papaveraceae

              Corn Poppy

              Pust-e shaghayegh

              Rind

              27.25 ± 0.12

              93.

              Papaver Rhoeas

              Papaveraceae

              Corn Poppy

              Gol-e shaghayegh

              Flower

              97.50 ± 0.01

              94.

              Papaver somniferum

              Papaveraceae

              Opium Poppy

              Khashkhash

              Seed

              4.79 ± 0.03

              95.

              Papaver somniferum

              Papaveraceae

              Opium Poppy

              Khashkhash

              Fruit

              35.95 ± 0.02

              96.

              Passiflora caerulea

              Passifloraceae

              Passion Flower

              Gol-e sâAty

              Flower

              46.90 ± 0.008

              97.

              Pelargonium graveolens

              Geraniaceae

              Geranium

              Barg-e atr

              Leaf

              92.19 ± 0.01

              98.

              Pelargonium graveolens

              Geraniaceae

              Rose Pelargonium

              Gol-e atr

              Flower

              96.87 ± 0.02

              99.

              Pterocarpus rubra

              Fabaceae

              Mukwa

              Sandal-e sorkh

              Bark

              91.75 ± 0.01

              100.

              Petroselinum hortense

              Apiaceae

              Parsley Seed

              Tokhm-e jafari

              Seed

              50.35 ± 0.03

              101.

              Pistachio lentiscus

              Anacardiaceae

              Lentisk Pistache

              Mastaki

              Gum

              92.37 ± 0.01

              102.

              Pistacia vera

              Anacardiaceae

              Pistachio Nut Shell

              Pust-e peste

              Rind

              97.71 ± 0.01

              103.

              Plantago major

              Plantaginaceae

              Great Plantain

              Bârhang

              Seed

              4.69 ± 0.11

              104.

              Polyporus officinalis

              Fomitopsidaceae

              White Agaric

              Ghariqun

              Fungi

              19.97 ± 0.06

              105.

              Portulaca oleracea

              Portulacaceae

              Common Purslane Seed

              Tokhm-e khorfe

              Seed

              7.19 ± 0.03

              106.

              Prunus persica

              Rosaceae

              Peach

              Barge-e holo

              Fruit

              9.47 ± 0.11

              107.

              Punica granatum

              Lythraceae

              Pomegranate Flower

              Golnar

              Flower

              99.90 ± 0.01

              108.

              Punica granatum

              Lythraceae

              Pomegranate

              Golnar

              Rind

              99.90 ± 0.01

              109.

              Quercus infectoria

              Fagaceae

              Oak Gall

              Qolqaf

              Gall

              53.97 ± 0.02

              110.

              Quercus infectoria

              Fagaceae

              Oak Fruit Hull

              Jaft

              Rind

              98.84 ± 0.02

              111.

              Rheum ribes

              Polygonaceae

              Rhubarb

              Rivâs

              Root

              98.93 ± 0.01

              112.

              Rosa centifolia

              Rosaceae

              Damask Rose

              Gol-e sorkh

              Flower

              97.51 ± 0.01

              113.

              Rosa foetida

              Rosaceae

              Rosa Lutea

              Gol-e zard

              Flower

              89.19 ± 0.023

              114.

              Rosmarinus angustifolia

              Lamiaceae

              Pine Rosemary

              Rozmary-e aklilaljabal

              Leaf

              22.51 ± 0.02

              115.

              Rubia tinctorium

              Rubiaceae

              Madder Root

              Ronas

              Root

              37.31 ± 0.02

              116.

              Ruta graveolens

              Rutaceae

              Rue

              Sodab

              Leaf

              27.91 ± 0.04

              117.

              Saccharum officinarum

              Poaceae

              Sugar Cane

              Shekar-e sorkh

              Mann

              35.04 ± 0.06

              118.

              Salix aegyptiaca

              Salicaceae

              Aegyption Willow

              Bidmeshk

              Flower

              17.05 ± 0.02

              119.

              Salix sp.

              Salicaceae

              Whitewillow

              Pust-e bid

              Bark

              38.10 ± 0.01

              120.

              Salvia hydrangea

              Lamiaceae

              Mountain Sage

              Gol-e arune

              Flower

              91.09 ± 0.01

              121.

              Salvia macrosiphon

              Lamiaceae

              Willd Sage Seeds

              Thokhm-e marv

              Seed

              2.86 ± 0.04

              122.

              Sambucus ebulus

              Adoxaceae

              Dwarf Elder

              Tarâsit

              Fruit

              99.70 ± 0.01

              123.

              Santalum album

              Santalaceae

              Sandalwood

              Sandal-e sefid

              Bark

              58.69 ± 0.02

              124.

              Satureja hortensis

              Lamiaceae

              Savory Seed

              Tokhm-e marze

              Seed

              35.77 ± 0.04

              125.

              Scrophularia striata

              Scrophulariaceae

              Striata Figwort

              Mokhallace

              Stem& Flower

              16.47 ± 0.05

              126.

              Sinapis alba

              Brassicaceae

              White Mustard

              Khardal-e sefid

              Seed

              39.77 ± 0.06

              127.

              Spinacia oleracea.

              Amaranthaceae

              Spinach Seed

              Tokhm-e esfenaj

              Seed

              19.76 ± 0.04

              128.

              Taraxacum sp.

              Asteraceae

              Dandelion

              Ghasedak

              Flower

              14.83 ± 0.01

              129.

              Thymus kotschyanus

              Lamiaceae

              Kotschyam Thyme

              Avishan

              Herb

              17.94 ± 0.01

              130.

              Tilia platyphyllos

              Malvaceae

              Linden

              Zirfun

              Leaf & Flower

              25.79 ± 0.01

              131

              Trigonella foenum-graecum

              Fabaceae

              Fenugreek Seed

              Tokhm-e shanbalile

              Seed

              44.02 ± 0.02

              132.

              Triticum sativum

              Poaceae

              Wheat

              Sabos-e ghandom

              Husk

              16.14 ± 0.04

              133.

              Tussilago farfara

              Asteraceae

              Colt′s-foot

              Pakhari

              Herb

              69.08 ± 0.01

              134.

              Veratrum album

              Melanthiaceae

              White Hellebore

              Kharbogh

              Leaf

              96.85 ± 0.06

              135.

              Verbascum georgicum

              Scrophulariaceae

              Mullein

              Dom-e gav

              Leaf

              30.40 ± 0.03

              336.

              Verbascum sp.

              Scrophulariaceae

              Mullein

              Marg-e mâhi

              Fruit

              0.82 ± 0.05

              137.

              Ziziphus vulgaris

              Rhamnaceae

              Jujube

              Annâb

              Fruit

              26.34 ± 0.01

              138.

              Hydroxyurea

              -------------

              ------------

              ----------

              Reference compound

              100 ± 0.01

              Table 2

              IC 50 and medicinal uses of most active plants

               

              Scientific name

              Effects & medicinal uses

              IC50(μg/ml)

              Std. Error log IC50

              1.

              A. catechu

              Anthelmintic, gastric tonic

              216

              0.01

              2.

              C. cristata

              Styptic, depurative, sedative, constipating, antibacterial, febrifuge,

              6175

              0.68

              3.

              C. annuum

              Anti flatulence, gout, gastric tonic, paralysis

              751

              0.14

              4.

              C. aurantifolia

              Appetitive, anti-flatulence, analgesic

              432

              0.06

              5.

              C. cassia

              Gastric tonic, anti-spasmodic, anti-flatulence

              867

              0.05

              6.

              C. microphylla

              Anti-flatulence, gastric tonic

              665

              0.14

              7.

              C. sinensis

              Anti-bacterial, anti diarrhea, diuretic, astringent, reduce cholesterol

              579

              0.04

              8.

              C. zeylanicum

              Gastric tonic, anti-flatulence, ‘anti-spasmodic

              361

              0.02

              9.

              C. sinensis

              Anti-diarrhea, diuretic, astringent, anti-bacterial, reduce cholesterol

              1314

              0.04

              10.

              Cetaurea sp.

              Anti-inflammatory, astringent, emmenagogue, sedative

              5152

              0.05

              11.

              H. gossypifolius

              Analgesic, anti-tussive, demulcent, diuretic, febrifuge, highly emollient, slightly laxative and odontalgic, anti-inflammations and laryngitis,

              819

              0.01

              12.

              H. perforatum

              Astringent, analgesic, anti-inflammator, anti-anxiety aphrodisiac

              3509

              0.10

              13.

              J. regia

              Anti-inflammatory, astringent, anti-spasmodic

              1271

              0.08

              14.

              M. chamomilla

              Anti-inflammation, appetitive, and aids digestion and sleep, acts as a diuretic and nerve tonic.

              3188

              0.02

              15.

              M. fragrans

              Anti-flatulence appetitive ‘anti-spasmodic’ antiseptic, analgesic, anti-inflammatory

              215

              0.15

              16.

              M. spicata

              Analgesic, Anti-spasmodic, anti-flatulence

              7822

              0.17

              17.

              M. communis

              Antiseptic, disinfectant, expectorant, deodorizer

              170

              0.04

              18.

              N. officinale

              Diuretic, expectorant, purgative, hypoglycemic, odontalgic, stimulant, tonic and stomachic

              2055

              0.19

              19.

              N. alba

              Astringent, antiseptic, anesthetic, aphrodisiac, sedative, used for gastrointestinal disorders and jaundice

              820

              0.19

              20.

              N. Oleander

              Dermatitis, abscesses, eczema, psoriasis, sores, warts, corns, ringworm, scabies, herpes, skin cancer, asthma, dysmenorrheal, epilepsy, malaria,

              9877

              0.26

              21.

              N. tabacum

              Anti-spasmodic, diuretic, sedative, sialagogue

              473

              0.15

              22.

              O. europea

              Hypotensive, diuretic, hypoglycemic

              2857

              0.06

              23.

              P. granatum(Rind)

              Hypoglycemic, anti-cancer, anthelmintic

              1484

              0.10

              24.

              P. granatum (Flower)

              Hypoglycemic, anti-cancer, anthelmintic

              1331

              0.11

              25.

              P. graveolens

              Anti-inflammatory, antiseptic, aromatherapy, astringent, anti- cancer, sedative

              976

              0.03

              26.

              P. graveolens

              Analgesic, anti-Bacterial, anti-Depressant, anti-inflammatory, antiseptic, astringent, diuretic, insect repellent, refreshing, relaxing, sedative, styptic, tonic

              1242

              0.14

              27.

              P. lentiscus

              Antibacterial

              121

              0.03

              28.

              P. Rhoeas

              Anodyne, emmenagogue, emollient, expectorant, hypnotic, sedative, tonic

              5636

              0.04

              29.

              P. rubra

              Astringent, tonic

              930

              0.06

              30.

              P. vera

              Aphrodisiac, anti-anxiety

              4687

              0.12

              31.

              Q. infectoria

              Gingivitis, infectoria, anti-diabetic, anti-tremorine, local anesthetic, antiviral, antibacterial, antifungal.

              1214

              0.12

              32.

              R. centifolia

              Anti-inflammatory, antispasmodic, aphrodisiac, astringent, depurative, laxative, analgesic, appetitive

              544

              0.07

              33.

              R. foetida

              Heart diseases, digestive, skin diseases, muscular pains, anti-parasite

              2441

              0.19

              34.

              R. ribes

              Appetitive, astringent, anti bacteria, anti depressive and used to treat diabetes, hemorrhoids, ulcer, diarrhea

              92

              0.06

              35.

              S. hydrangea

              anti-flatulence, astringent, anti-spasmodic

              2960

              0.11

              36.

              S. ebulus

              Anti-inflammatory, antinociceptive, anti-cancer, anti-angiogenic, anti-oxidative

              57

              0.05

              37.

              V. album

              Analgesic, anthelmintic, cathartic, emetic, expectorant, hypnotic

              1037

              0.07

              38.

              hydroxyurea

               

              37

              0.02

              S. ebulus (Figure 1) is a native perennial herb of the Adoxaceae family [19]. It has been prescribed in traditional medicines for the treatment of inflammatory reactions, such as hemorrhoid, bites and sore-throat. In addition, S. ebulus has been shown to have anti-inflammatory, antinociceptive, anti-cancer, anti-angiogenic and anti-oxidative activities. Ebulitin, ebulin 1, flavonoid, anthocyanin and other components have been isolated from S. ebulus and identified as active ingredients of biological and pharmacological activities [20]. The anti-H. pylori effect of the S. ebulus extract was observed by using the agar dilution method [13].
              http://static-content.springer.com/image/art%3A10.1186%2F2008-2231-20-72/MediaObjects/40199_2012_Article_72_Fig1_HTML.jpg
              Figure 1

              Sambucus ebulus in flowering stage[19].

              R. ribes (Figure 2) is a hardy perennial, cultivated in some temperate countries for its edible red leaf stalks [21]. It is used to treat diabetes, hemorrhoids, ulcer, diarrhea, and expectorant activity reported. The efficacy and safety of a hydroalcohlic extract of R. ribes in treatment of mild to moderate major depression disorder has been investigated and the observations show some anti depressive effects. The methanolic extract of R. ribes have demonstrated anti-ulcer activity comparable with standard drugs cimetidine [22].
              http://static-content.springer.com/image/art%3A10.1186%2F2008-2231-20-72/MediaObjects/40199_2012_Article_72_Fig2_HTML.jpg
              Figure 2

              Rheum ribes leaf[21].

              According to strong inhibitory activity of the herbs presented in Table 2, simultaneous application of theses herbs and the medicines prescribed in gastrointestinal disease therapies would fasten the treatment. Additionally, isolation of active compounds and further investigation of each isolated compound against urease activity would lead to new chemical structures which may have the potency to inhibit urease activity.

              Declarations

              Acknowledgements

              We would like to thank Fereshteh Keyghobadi for her cooperation in the practical herbal testes. The financial supports of the Research Council of the Tehran University of Medical Science, University of Tehran and Iran National Science Foundation (INSF) are gratefully acknowledged.

              Authors’ Affiliations

              (1)
              Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences
              (2)
              Department of Pharmacognosy, School of Pharmacy, Shahid Beheshti University of Medical Sciences
              (3)
              School of Biology, College of Science, University of Tehran
              (4)
              Medicinal Plants Research Center, Tehran University of Medical Sciences
              (5)
              School of Advanced Medical Technologies, Tehran University of Medical Sciences

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              Copyright

              © Nabati et al.; licensee BioMed Central Ltd. 2012

              This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://​creativecommons.​org/​licenses/​by/​2.​0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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