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有梗石龙尾甲醇提取物抗志贺菌病作用的体内和体外研究(英文)



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DOI:10.3736/jcim20120509
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Subhadra S,Gade G,Ravindran VK,Emani VS,
Parre S,Banji D.In vitro and in vivo evaluation of the
inhibitory effect of Limnophila indica (Linn.)Druce
on shigelosis.J Chin Integr Med.2012;10(5):538-
545.
Subhadra S,Gade G,Ravindran VK,Emani VS,
Parre S,Banji D.有梗石龙尾甲醇提取物抗志贺菌病
作用的体内和体外研究.中西医结合学报.2012;10
(5):538-545.
Received November 11,2011;accepted January 6,
2012;published online May 15,2012.
Ful-text LinkOut at PubMed.Journal title in PubMed:
Zhong Xi Yi Jie He Xue Bao.
Correspondence:Sandhya Subhadra,PhD;Tel:+91-
90-10055005;E-mail:sanpharm@gmail.com
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Original Experimental Research实验论著
In vitro and in vivo evaluation of the inhibitory effect
of Limnophila indica(Linn.)Druce on shigelosis
Sandhya Subhadra,Gouthami Gade,Vinod Kombath Ravindran,Vidya Sravanthi Emani,
Saikumar Parre,David Banji
Department of Pharmacognosy,Nalanda Colege of Pharmacy,Cherlapaly,Nalgonda 508001,Andhra
Pradesh,India
OBJECTIVE:The research aimed to evaluate the anti-Shigela and antacid activities of the
methanolic extract of Limnophila indica.
METHODS:The whole plant of L.indica was extracted using methanol and then subjected to
preliminary chemical screening.The in vitro antibacterial screening on two Gram-positive and
two Gram-negative bacteria as wel as three Shigelaspecies of which two bacteria were
antibiotic-resistant were evaluated by disc difusion method.Castor oil-induced diarrhoea on
Wistar albino rats was performed by using loperamide as a standard control.The in vitro
antacid activity was tested by artificial stomach model.The neutralization eficiency,capacity,
volume and hydrogen ions consumed were also evaluated.
RESULTS:The preliminary chemical screening on methanolic extract of L.indica showed the
presence of phenolic compounds,flavonoids,alkaloids,fats and oils.It was proved to be a
potent antibacterial agent against the four bacterial strains.Screening against Shigela species
revealed that it was a powerful antibacterial agent towards antibiotic-resistant Shigela
species.In the case of in vivo antidiarrheal activity,the plant has shown a dose-dependent
activity and the lowest dose at 100 mg/kg gave a much better result than loperamide(P<
0.01).The in vitro antacid study showed a mild activity.
CONCLUSION:As the plant L.indica has been proved to be a competent antibacterial as
wel as a compeling antidiarrheal agent with mild antacid activity,this plant can be very wel
suggested to be an eminent substitute for the various synthetic anti-dysentery and antidiarrheal
agents available in the market.
KEYWORDS:Limnophila;Shigela dysenteriae;antidiarrheals;antacids;dyspepsia;rats
·835· 中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
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 Dysentery epidemics have worsened the burden
of endemic shigelosis in developing countries.
Approximate 140 milion cases and576 000 deaths
occur annualy due to Shigela infection in children
under five years old worldwide[1].Bacterial dysentery
is the most widespread and acute infectious enteritis
of human and nonhuman primates caused by bacteria
of the genus Shigella.It usualy causes recurrent
bloody mucoid stools accompanied by abdominal
cramps and rectal pain.Life-threatening problems of
shigelosis are haemolytic uremic syndrome,
encephalopathy,colonic ulceration and perfora-
tion,shigelemia,toxic megacolon,intestinal
stenosis and obstruction,persistent diarrhea and
severe malnutrition[1].Treatment includes the usage
of synthetic drugs such as antibiotics and anti-
dysenteric drugs[2].Plants are imperative source
of potentialy useful structures for the expansion
of new therapeutic agents in which the initial step
is in vitro antibacterial activity[3].Limnophila
indicais an aquatic or marshy submerged,aromatic,
short,erect diffusely branched herb which belongs
to the family Scrophulariaceae.The identified chemical
constituents of the plant include flavonoids like
(2S)-5,7,3′,4′-tetramethoxyflavanone and 5,7,2′,5′-
tetramethoxyflavone,7-omethylwogonin,skulcapflavon
1,5-hydroxy-7,20-dimethoxyflavone and hydroxy-
6,8-dimethoxy-3,4-methylenedioxyflavone[4].The
traditional medical characteristics of the plant
include carminative,antiseptic,anti-filariasis,anti-
dysentery and anti-dyspepsia activities
[5].Since
the bacterial dysentery is a major epidemic which
affects al age groups and most of the bacteria are
prone to antibiotic resistance,the need of identifying
the potent antibacterial compounds has become
obligatory.L.indica is a traditionaly important
plant which is said to be a potent and unique plant
used by the folklore of Indian subcontinent for
dysentery and dyspepsia.The present research
exploration was consequently undertaken to evaluate
the anti-Shigella activity by in vitro and in vivo
studies.Furthermore,the in vitro antacid screening
was also performed on artificial stomach model to
determine its antacid potency.
1 Materials and methods
1.1 Plant materials and preparation Plant was
colected from the forest region of Thalakona in
Chittoor district,India and was authenticated by
retired Professor A.Laxma Reddy,Department of
Botany,Nagarjuna Government Colege,Nalgonda,
India.The plant herbarium was prepared and
deposited in the Department of Pharmacognosy,
Nalanda Colege of Pharmacy,India for future
reference.The plant was identified as Limnophila
indica (Linn.)Druce(family Scrophulariaceae)
(Voucher No:NCOP-NLG/phcog/10-11/038).
1.2 Procurement of microorganisms The bacterial
strains Pseudomonas aerugenosa MTCC741,Bacilus
subtilis MTCC441,Escherichia coli MTCC443,
Staphylococcus aureus MTCC 96 and Shigella
flexneri MTCC1457 were procured as freeze-dried
form from the Microbial Type Culture Colection,
Chandighar,India.Shigella dysentery and Shigela
boydii were the two pathogenic antibiotic-resistant
bacterial strains procured from the Microbiology
Laboratory of Kamineni Institute of Medical
Sciences,Nalgonda,Andhra Pradesh,India.
1.3 Animals Wistar albino rats of either sex
weighing 150to200 g were procured from National
Institute of Nutrition,Hyderabad,Andhra Pradesh,
India.The experimental protocol was approved
by the institutes animal ethics committee under
the reference No. NCOP/IAEC/approval/36/
2011 and the experimental studies were conducted
according to their rules and regulations.The animals
were housed in metabolic cages bedded with rice
husk under standard environmental conditions and
had free access to standard pelet diet and water
ad libitum.
1.4 Plant extraction and preliminary chemical
screening The powdered plant material weighing
500 g was defatted with petroleum ether and then
extracted with methanol for 72 h at 45 ℃.The
obtained extract was then concentrated under
vacuum using rotary vacuum evaporator and subjected
·935·中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
to preliminary chemical screening to identify the
active chemical constituents[6,7].
1.5 In vitro anti-Shigella activity An initial
screening of antibacterial activity was performed
on two Gram-positive organisms,B.subtilis and
S.aureus,and two Gram-negative organisms,
P.aerugenosa and E.coli to evaluate the anti-
bacterial potency of the plant.Kirby-Bauers
method[8 ]was folowed using the standardized
sterile paper disc agar diffusion method.Al Petri
dishes and graduated measuring pipettes were
heat-sterilized in an autoclave at 120℃for 1 h.
Media were steam-sterilized at 121℃ (103 kPa)
for 20 min in an autoclave.Al plates were prepared
with equal thickness of nutrient agar.Concentra-
tions of 25,50,75 and 100 mg/mL solutions of
methanol crude extracts of L.indica were
prepared and 30μL of each test solution was
incorporated into the sterile disc,and then placed
on the solidified nutrient agar plates inoculated
with the microorganisms.Gentamycin(1 mg/mL)
and methanol were employed as standard drug and
normal control groups,respectively.The plates
were then incubated at 37 ℃ for 24 h.Al the
tests were performed in triplicates and the results
were expressed as mean of the inhibition
diameters (mm)[9].The in vitro anti-Shigella
screening against S.flexineri,S.dysentery and
S.boydii was also performed as per the above
mentioned procedure.In this work,instead of the
standard drug gentamycin,ceftazidime(1 mg/mL)
was selected as the reference drug since S.dysentery
and S.boydii were resistant to gentamycin.
1.6 Determination of minimal inhibitory concen-
tration The minimal inhibitory concentration
(MIC )was determined as per Shafi[10],with
modifications.The stock solution with 1 mg/mL
concentration was prepared and the serial dilutions
were prepared and incorporated into the discs.
These discs were dried and placed in the solidified
nutrient agar media inoculated with the test
organisms P.aerugenosa,E.coli,B.subtilis,
S.aureus,S.flexneri,S.dysentery and S.boydii.
The size of the inoculums used was approximately
20 microorganisms,and then incubated at 37℃for
24 h.
1.7 Safety evaluation The safety evaluation
studies were performed on female nuliparous and
non-pregnant Swiss albino mice.Each animal was
between 8 to 12 weeks old and the weight was 25
to 30 g.Twelve animals were taken and divided
into four groups with three in each group.The
animals in groupⅠto groupⅣ were respectively
given the methanol extract of 5,50,300 and
2 000 mg/kg oraly in single dose by gavages using
stomach tube.The animals were then kept under
observation and were frequently checked for alergic
reactions,toxic symptoms and abnormalities for
one week[11].
1.8 In vivo antidiarrheal activity The in vivo
antidiarrheal screening by castor oil-induced
diarrhea in Wistar albino rats was performed.
Animals were housed individualy with free access
to food and water under standard condition and
the basal food intake,and then the animals were
starved for 18 h prior to starting the biological
activity test.The rats were grouped into four
groups with six in each according to their weights
and housed in separate localy fabricated metabolic
cages.Group Ⅰ was designated as the normal
control receiving normal saline(1 mL/kg),group
Ⅱreceived 3 mg/kg loperamide as the standard,
groupsⅢandⅣ were given graded doses of 100
and 200 mg/kg (per oral)methanol extract of
L.indica,respectively.Al the samples were
given 1 h prior to the oral administration of
cathartic agent of 2 mL castor oil.Oral adminis-
tration of castor oil was facilitated by the use of a
stomach tube.The animals were monitored for
12 h for the consistency of stool and the frequency
of defecation.At the end of this period,the total
number of faecal matter for each group and the
number of diarrhoeic (wet)faeces which were
recorded and the mean value for each group were
calculated[12,13].
1.9 In vitro antacid screening by modified artificial
stomach model This work was performed on
indigenously self-fabricated modified artificial
stomach model.The 136 air bubbles per minute
were used to mimic the peristaltic movements of
the stomach.
1.9.1 Preparation of artificial gastric juice Two
grams of salt and 3.2 mg of pepsin enzymes were
dissolved in 500 mL of water,and then 7 mL of
hydrochloric acid and adequate water were added
to make a 1 000 mL solution of the artificial
gastric acid at pH1.2[14].
1.9.2 Effect of temperature on pH About 90 mL
of extract were used to determine the effect of
temperature on the pH of the artificial gastric
acid.The temperatures taken into account were
at 29℃ (room temperature)and 37℃ (human
body temperature).The pH values of the control
solution (water)and sodium bicarbonate were
also determined.
1.9.3 Neutralizing effect of extract on artificial
gastric acids Totaly 90 mL of each test solution
were added to 100 mL of artificial gastric juice at
pH1.2.The pH value was determined to examine
the neutralizing effect
[14].
1.9.4  Determination of the duration of the
neutralizing effect Totaly 90 mL of each test
solution were added to 100 mL of the artificial
gastric juice at pH1.2 in the container of the
artificial stomach at 37 ℃ and the aerator was
arranged with 136 air bubbles per minute.Artificial
gastric juice at pH 1.2 was pumped into the
container of artificial stomach at 3 mL/min,and
it was pumped out at 3 mL/min at the same time.
A pH meter was connected to continuously
·045· 中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
monitor the changes of pH in the container of the
artificial stomach.The duration of neutralization
effect was determined when the pH value was
returned to its initial of pH1.2[14].
1.9.5 Determination of neutralizing capacity by
Fordtrans in vitro titration model A total of
90 mL of the test sample were placed in 250 mL
of beaker and warmed to 37℃.An aerator was
arranged with 136 air bubbles per minute to imitate
the stomach movements.The test samples were
titrated with the artificial gastric juice to the end
point of pH3.0.The consumed volume (V)of
the artificial gastric juice was measured.The total
consumed hydrogen ion (mmol)was measured as
0.063 096 mmol/mL×V (mL)[15].
1.10 Statistical analysis Data were expressed as
mean±standard deviation.The difference between
experimental groups was compared by one-way
analysis of variance folowed by Dunnetts t-test.
The software of Graph Pad Instat was used for
analyzing the multiple comparison tests.The
differences were considered to be statisticaly
significant when P<0.01.
2 Results
2.1 Plant methanolic extraction and preliminary
phytochemical screening The yield of methanolic
extract obtained from500 g of the dried powdered
plant of L.indica was found to be 14.5% weight
ratio.When this crude extract was subjected to
preliminary chemical screening,it showed the
presence of phenol compounds,flavonoids,alkaloids,
fats and oils.
2.2 In vitro anti-Shigelaactivity The antibacterial
studies revealed that the methanol extract of
L.indica was a potent antibacterial agent.It was
observed that the extract at 100 mg/mL showed
more potent antibacterial activity than gentamycin at
1 mg/mL against S.aureus and E.coli.The
extract has shown similar antibacterial potency as
that of gentamycin with the bacterial strain
P.aerugenosa and B.subtilis(Table 1).
 L.indica showed an excelent anti-Shigella
activity.It was observed that S.flexineri was
found to be highly sensitive to the extract than the
standard drug among the three Shigella species.
The zone of inhibition obtained for the extract at
100 mg/mL was significantly larger than that of
the ceftazidime at 1 mg/mL.S.boydii was found
to be almost equaly susceptible to the methanol
extract at 100 mg/mL as wel as ceftazidime.While
S.dysentery was found to be least vulnerable to the
methonal extract (Table 2).Compared with the
normal control group,both the methanol extract
of L.indica and ceftazidime were found to be more
anti-Shigela (P<0.01).
2.3 MIC of the methanol extract of L.indica 
The MIC revealed that 230,233 and 235μg/mL
were the minimum concentrations of the plant
extract required to inhibit the growth of S.flexinari,
S.dysentery and S.boydi respectively.The extract
can be categorized as moderately active as per
(Table 3)MIC value obtained for P.aerugenosa
and S.aureus while the value obtained for E.coli
and B.subtilis (Table 4)fel under the category
of weakly active.
Table 1 Antibacterial activity of methanol extract of Limnophila indica
(Mean±standard deviation)
Group  n
Zone of inhibition(mm)
Bacillus subtilis
Staphylococcus
aureus
Pseudomonas
aerugenosa
Escherichia coli
Normal control  3  0.54±1.00  1.20±2.00  1.50±1.53  0.85±0.58
Gentamycin(1mg/mL) 3  15.67±1.16** 14.67±0.58** 14.67±0.58** 15.00±0.00**
Limnophila indica methonal extract(25mg/mL ) 3  1.67±1.16** 11.67±1.53** 11.67±1.53** 9.33±1.53**
Limnophila indica methonal extract(50mg/mL ) 3  7.33±1.16** 12.00±0.00** 11.67±0.58** 11.67±0.58**
Limnophila indica mehonal extract(75mg/mL ) 3  10.67±1.16** 13.00±1.00** 12.67±1.16** 14.33±0.58**
Limnophila indica methonal extract(100mg/mL ) 3  12.00±2.00** 15.00±1.00** 14.67±1.16** 20.00±1.00**
  **P<0.01,vs normal control group.Sterile discs were loaded with 30μL/disc of respective compounds.
Table 2 In vitro anti-Shigellaactivity of methanol extract of Limnophila indica
(Mean±standard deviation)
Group n
Zone of inhibition(mm)
Shigella flexneri  Shigella dysentery  Shigella boydii
Normal control  3  0.68±0.58  1.00±0.58  0.62±1.00
Ceftazidime(1mg/mL) 3  15.00±0.00** 28.00±1.00** 20.00±0.00**
Limnophila indica methonal extract(25mg/mL ) 3  11.33±0.58** 11.00±0.00** 10.33±0.58**
Limnophila indica methonal extract(50mg/mL ) 3  12.33±0.58** 12.33±0.58** 12.33±0.58**
Limnophila indica mehonal extract(75mg/mL ) 3  14.33±0.58** 14.66±0.58** 14.33±0.58**
Limnophila indica methonal extract(100mg/mL ) 3  18.33±0.58** 18.67±0.58** 19.00±0.00**
  **P<0.01,vs normal control group.Sterile discs were loaded with 30μL/disc of respective compounds.
·145·中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
Table 3 Minimum inhibitory concentration for grading
the methanol extract of Limnophila indica
Minimum inhibitory concentration(μg/mL ) Grade
≤20 More active
20to 100 Active
100to 250 Moderately active
250to 500 Weakly active
>500 Inactive
Table 4 MIC value obtained for the antimicrobial activity of
methanol extract of Limnophila indica
Microorganism  MIC(μg/mL)
Bacillus subtilis  335
Staphylococcus aureus  180
Pseudomonas aerugenosa  190
Escherichia coli  225
Shigella flexneri  230
Shigella dysentery  233
Shigella boydii  235
  MIC:minimum inhibitory concentration.
2.4 Safety evaluation Safety evaluation revealed
that plant extract was safe since none of the
animals showed any kind of toxic symptoms or
behavior changes up to the dose of 2 000 mg/kg.
2.5 In vivo antidiarrheal activity The antidiarheal
activity in castor oil-induced model revealed that
the methanol crude extract of L.indica possessed
substantial antidiarrheal activity. Upon 12 h of
observation,the complete cessation of wet motions
was seen in animals treated with 200 mg/kg of
methanol extract by 6 h.The consistency of wet
motion was found to be semisolid in nature for the
animals in 200 mg/kg of methanol extract group.
In the case of animals treated with 100 mg/kg
methanol extract of L.indica,the latency period
of passing the motion had increased drasticaly,
while the complete cessation of wet motions was
not observed even after 12 h.The consistency of
motion in the first three hours of observation
revealed the watery nature,but later the frequency
of passing the motion reduced and the consistency
turned to semisolid nature.The animals treated with
standard drug of loperamide produced watery loose
motions in the initial 3 h and later the consistency
changed to semisolid nature.This group of animals
did not show100%termination of wet motions.
The animals treated with normal saline were
found to be in complete diarrheic condition til the
10th hour,while the later condition of motion improved
to semisolid nature(Table 5).
Table 5 Antidiarrheal property of methanol extract of Linmophila indica
(Mean±standard deviation)
Group n  Total number of defecation  Number of wet defecation
Control(1mL/kg saline) 6  11.40±0.24  8.00±0.32
Standard(3mg/kg loperamide ) 6  2.20±0.58△△ 0.60±0.24△△
Limnophila indica methanol extract(100mg/kg) 6  1.60±0.40△△ 0.20±0.20△△
Limnophila indica methanol extract(200mg/kg) 6  0.80±0.37△△ 0
  △△P<0.01,vs control group.
2.6 In vitro antacid screening by modified artificial
stomach model
2.6.1 Effect of temperature on pH The pH
values of the test concentrations,standard and
control at 29 and 37 ℃ were determined to
recognize the effect of temperature on pH.It was
observed that there were no alterations in the pH.
2.6.2 Neutralizing effect of the extract on the artificial
gastric acids The plant extract at 200 mg/kg was
found to increase in pH compared with the control
group (P<0.01).The standard drug of sodium
bicarbonate at 100 and 200 mg was determined to
be highly efficient compared with the normal
control group(P<0.01).See Figure 1.
2.6.3 Duration of neutralizing effect tested by
modified Vatiers artificial stomach model The
time taken by 100 and 200 mg/kg of methanol
crude extract of L.indica to neutralize the pH
showed that the extract had a moderate activity.
The sodium bicarbonate at 200 mg/kg exhibited
an excelent duration as compared with the control
group (P<0.01).See Figure 2.
2.6.4 Determination of neutralizing capacity by
Fordtrans in vitro titration model The volume of
artificial gastric juice consumed by the plant
extract was decreased compared with the sodium
bicarbonate,which revealed that the plant had a
mild antacid activity.The crude extract at 200 mg/kg
consumed much less than that of the sodium bicar-
bonate at the same concentration (P<0.01,see
Figure 3).Numbers of hydrogen ion consumed by
200 mg/kg of the plant extract showed that the
plant is mildly antacid(Figure 4).
Figure 1 Neutralization effect of methanol extract of
Linmophila indica on artificial gastric juice
C:Control group;S1:100mg/kg sodium bicarbonate group;S2:
200mg/kg sodium bicarbonate group;E1:100mg/kg Limnophila
indica methanol extract group;E2:200mg/kg Limnophila indica
methanol extract group.Data are expressed as mean±standard
deviation;n=6;▲▲P<0.01,vs control group.
·245· 中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
3 Discussion
 Shigelosis is a disease that affects a large proportion
of human population especialy the children.The
increase in antibiotic resistance of microorganism
against conventional drugs has necessitated the
search for new,efficient and cost-effective ways
for the control of infectious diseases.The results of
different studies provided evidence that some
medicinal plants might be a potential source of new
antibacterial agents[15]. L.indica is a potent
medicinal herb used by the folklore of India for
the treatment of dysentery,diarrhoea,pestilent
fever and dyspepsia.Hence in this context the anti-
dysenteric property was evaluated to establish its
traditional use scientificaly.The in vitro anti-
bacterial studies on routine bacterial strains as
wel as three Shigellaspecies revealed that the
methanol extract is a persuasive inhibitor of these
microorganisms.Interestingly,it was observed
that the plant extract produced a potent cidal
effect on the two bacterial strains namely
S.dysentery and S.boydii which were procured as
antibiotic-resistant strains.The chemical screening
of the methanol extracts of whole plant showed
the presence of phenol compounds,flavonoids,
alkaloids,fats and oils.This antibacterial potency
of L.indica can be attributed to the chemical
entities either singly or in combination by producing a
synergetic effect.Phenol compounds are thought
to be toxic to microorganisms,inhibiting the
enzymes which are essential for the growth of
microorganism.The literature survey revealed the
phenol-or polyphenol-like flavonoids to be potent
antimicrobial agents.The modes of action exhibited
by these compounds were degrading of the cel
wal,interacting with the composition and disrupting
the cytoplasmic membrane,damaging membrane
protein,interfering with membrane-integrated
enzymes,causing leakage of celular components,
coagulating cytoplasm,depleting the proton motive
force,changing fatty acid and phospholipids
constituents,impairing enzymatic mechanism for
energy production and metabolism,and altering
nutrient uptake and electron transportation[16,17].
Alkaloids are wel established for the antibacterial,
antifungal and antiparasitic properties.Approximately
300 alkaloids are reported to be antimicrobial in
nature[18].From the in vitro antibacterial studies it
was proved that L.indica possessed an excelent
antibacterial activity against Gram-positive and
Gram-negative organisms.Further investigation on
three Shigela species like S.flexinari,S.dysentery
and S.boydii revealed that the methanol extract
is a potent inhibitor of these microorganisms.
Hence further work on in vivoantidiarrheal property
was performed to explore the probable antidiarrheal
activity.Castor oil induces diarrhoea due to the
presence of ricinoleic acid.This acid with the
help of lipase irritates the intestinal mucosa and
produces inflammation with the release of prostaglandin
and nitric oxide that stimulate motility and secrete
electrolyte and water[19].It was observed that
·345·中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5
L.indica exhibited a more significant antidiarrheal
activity than the standard drug of loperamide.
The plant has shown a dose-dependent activity.It
was observed that the lower dose of 100 mg/kg
exhibited a beter antidiarrheal activity than loperamide
at 3 mg/kg.Since ricinolic acid produces hypersecretory
response,it can be aleged that the antidiarrheal
potency of the methanol extract of L.indica may
be through this anti-secretory mechanism.It is
wel documented that tannins,flavonoids,alkaloids,
saponins,reducing sugar,sterols and terpenes are
potent chemical entities with persuasive antidiarrheal
activity.These constituents are present in the
extract and may be responsible for this potent
pharmacological effect.It is wel recognized that
taking loperamide may cause a lot of side effects
like drowsiness,dizziness,fatigue,constipation
and vomiting.Moreover,this medicament is not
recommended for children under two[20].Plant-based
compounds have enormous therapeutic potential
as they can serve the purpose without any side
effects that are often associated with synthetic or
semisynthetic compounds.Hence it can be suggested
that L.indica can be a suitable alternative for
loperamide as it can be easily taken by any age
group.Acute dysentery or diarrhoea may often
lead to acidity or dyspepsia.Hence the in vitro
antacid study was performed on artificial stomach
model,and the results revealed that the plant
extract possessed mild antacid activity.The methanol
extract of the whole plant of L.indicashowed mild
neutralization efficiency with less duration of
neutralizing effect.The neutralizing capacity
with respect to the total volume of gastric juice
and hydrogen ions consumed was found to be very
less when compared with the standard drug of
sodium bicarbonate.
4 Conclusion
 As the plant was determined to be a strong
antibacterial drug against antibiotic-resistant organisms
as wel as a strong antidiarrheal drug compared
with the standard drug of loperamide which is
reported to have several side effects,it suggested
that this plant can be a potent substitute for synthetic
antidiarrheal and anti-dysentery drugs.
5 Acknowledgements
 The authors are thankful to the management of
Nalanda Colege of Pharmacy for providing al the
facilities in the library as wel as in the laboratory.
They express their deep sense of gratitude to
MTCC,Chandigarh and Microbiology Laboratory,
Kamineni Institute of Medical Sciences,Nalgonda,
India for providing the microbial cultures to perform
their research work.
6 Competing interests
 The authors declare that they have no competing
interests.
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有梗石龙尾甲醇提取物抗志贺菌病作用的体内和体外研究
Sandhya Subhadra,Gouthami Gade,Vinod Kombath Ravindran,Vidya Sravanthi Emani,Saikumar Parre,
David Banji
Department of Pharmacognosy,Nalanda Colege of Pharmacy,Cherlapaly,Nalgonda 508001,Andhra Pradesh,India
目的:探讨有梗石龙尾甲醇提取物抑制志贺菌病的作用。
方法:首先将有梗石龙尾甲醇提取物做初步化学筛选。使用2种革兰阴性菌、2种革兰阳性菌,2种抗生素耐
药的志贺杆菌和1种非抗生素耐药的志贺杆菌经纸片扩散法进行体外抗菌筛选。蓖麻油诱发 Wistar大鼠
腹泻后,以洛哌丁胺作为标准对照药进行有梗石龙尾甲醇提取物止泻实验。通过人工胃模型测定有梗石龙
尾甲醇提取物的体外抗酸作用,并且对其中和胃酸的效率、容量、体积和氢离子消耗进行评估。
结果:有梗石龙尾甲醇提取物中含有酚类、黄酮类、生物碱等化学成分,以及脂肪和油脂。有梗石龙尾甲醇提
取物被证明对包括抗生素耐药的志贺杆菌在内的4个菌株具有较强的抑制作用,并且其抗腹泻作用具有剂
量依赖性。当服用最低剂量100mg/kg时,其抗腹泻效果比标准对照药洛哌丁胺更好(P<0.01)。体内实
验结果显示有梗石龙尾甲醇提取物具有轻微的抗酸作用。
结论:有梗石龙尾甲醇提取物具有轻微的抗酸活性以及强效的抗菌、止泻作用,这对于开发抗痢疾、抗腹泻作
用的草药具有一定的意义。
关键词:石龙尾属;志贺菌,痢疾;止泻药;抗酸剂;消化不良;大鼠
·545·中西医结合学报2012年5月第10卷第5期 Journal of Chinese Integrative Medicine,May 2012,Vol.10,No.5