In vivo, in vitro, and in silico evaluation of the analgesic, antidiarrheal, and anthelmintic activities of methanolic extract of Syzygium grande (Wight) Walp

Objective: The present study was undertaken to evaluate the possible analgesic, neuropharmacological, anti-diarrheal, and cytotoxic activities of the ethanol extract of leaves of Solanum sisymbriifolium Lam. (Family: Solanaceae). Materials and Methods: The analgesic activity was measured by acetic acid-induced writhing inhibition test. The neuropharmacological activities were evaluated using hole cross, hole board, and elevated plus-maze test and the anti-diarrheal activity was assessed using castor oil-induced diarrhea inhibition method. Brine shrimp lethality bioassay was carried out for assessing the cytotoxicity of the ethanol extract of the leaves. Except cytotoxic activity, all the tests were conducted on mice.Results: The extract at oral doses of 200 and 400 mg/kg body weight showed highly significant (p<0.001) decrease in number of writhing, 52.1±0.66 and 4.4±0.64 compared with the control (78.6±0.29) with the percentage of inhibitions of writhing response were found to be 33.72% and 94.40%, respectively. Compare with the control, the extract at both doses showed significant sedative effect in hole cross test. In hole board test, the extract exhibited highly significant (p<0.001) anxiolytic activity at dose of (200 mg/kg), while the same activity was observed at dose of 400 mg/kg in elevated plus-maze test. The extract showed highly significant (p<0.001) anti-diarrheal activity in a dose-dependent manner. With the extract, significant lethality to brine shrimp was found with LC50 value of 61.66±0.9 μg/ml, which was comparable with the positive control (LC50: 11.89±0.8 µg/ml).Conclusion: The results from the present studies support the traditional uses of this plant part and could form the basis of further investigation including compound isolation.

The aim of this study was to phytochemically investigate the ethanolic extract of Hibiscus sabdariffa Linn. calyces and to evaluate the analgesic, anti-microbial and anti-diarrheal activities of this. The calyces of Hibiscus sabdariffa were separated from the other plant parts and sun dried and extracted using ethanol and phytochemically and pharmacologically evaluated.Different Phytochemical tests were performed for phytochemical screening and for determining the functional groups. Different methods like Disc diffusion method for antimicrobial activity determination, Castor oil induced diarrhea in mice method for the antidiarrheal study and Acetic acid induced writhing test for analgesic activity were employed. Phytochemical screening of the calyces of Hibiscus sabdariffa ensured the presence of alkaloid, flavonoids, saponins, tannins in the crude ethanolic extract. The peripheral analgesic activity was evaluated by acetic acid induced Writhing method. The extract produced 66.85% (p&lt; 0.001) inhibition of writhing in mice at the dose of 500-mg/kg body weight, which is comparable to diclofenac sodium (78.45% (p&lt;0.001) at the dose of 25mg/kg). The anti-diarrheal activity of the crude extract of Hibiscus sabdariffa was evaluated using the model of castor oil induced diarrhea in mice. The crude etanolic extract of Hibiscus sabdariffa (calyces) showed a marked antidiarrhoeal activity at dose of 500 mg/kg-body weight as compared to the standard antidiarrhoeal agent loperamide (dose:50mg/kg-body weight). Hibiscus sabdariffa caused an increase in latent period i.e. delayed the onset of diarrhoeal episode and decreased the frequency of defecation. Anti-microbial activity was tested using a number of micro-organisms. The peripheral analgesic activity of the ethanolic extract of the calyces of Hibiscus sabdariffa against acute inflammatory pain was significantly high as compared to potent inhibitory activity of Diclofenac (25mg/kg). Therefore, it is likely that the ethanolic extract at a dose of 400mg/kg might suppress the formation of these substances or antagonize the action of these substances and thus exerts its analgesic activity in acetic acid-induced writhing test. Anti-diarrheal activity was present in the ethanolic extract (500mg/kg) which indicate that the drug in decreased intestinal motility. This plant sample didn’t show any antimicrobial activity. DOI: http://dx.doi.org/10.3329/ijpls.v1i3.12978 International Journal of Pharmaceutical and Life Sciences Vol.1(3) 2012

Aeschynomene aspera (Family- Fabaceae), a traditionally used medicinal plant, is aquatic, perennial, erect subshrub up to 200 cm tall with compound leaves and locally it is called Shola (Bangladesh), Laugaun (India). The study started with phytochemical screening of ethanolic leaf extract of A. aspera followed by bioactivity study such as in-vitro antioxidant activity by free radical (DPPH) scavenging assay, estimation of total phenolic as well as total flavonoid contents and the analgesic, antidiarrheal and anthelmintic activities. Analgesic, antidiarrheal and anthelmintic activities were evaluated by acetic acid induced writhing inhibition in mice, castor oil induced diarrheal episode in mice and using living parasites Haemonchus contortus, respectively. The presence of carbohydrate, reducing sugar, glycosides, tannins, alkaloids, steroids, gums and flavonoids was estimated with the help of phytochemical screening. In DPPH scavenging assay the extract showed IC50 value of 86.14μg/mL where ascorbic acid showed 12.02μg/mL. It also exhibited total phenolic and flavonoid contents as 461.09 GAE/100g and 297.20mg Quercetin/100g of dried plant extract. About 36% and 61% writhing inhibitions were observed in mice at 250 and 500mg/kg doses of A. aspera leaf extract, respectively. Anthelmintic activity showed by the extract was dose dependent. The extract also showed 46.01 and 71.68% inhibition of defecation in mice at doses 250 and 500 mg/kg, respectively. All these suggest that the A. aspera extract possesses antioxidant, analgesic, anthelmintic and antidiarrheal activities. Bangladesh Pharmaceutical Journal 23(2): 109-116, 2020

Syzygium genus is extensively used in managing different diseases such as antimicrobial, anti-diarrheal, antidiabetic, analgesic, anti-inflammatory, antioxidant, and anticancer activities. The present study aims to investigate phytochemicals by screening tests and GC-MS analysis and evaluate the antioxidants, cytotoxicity, analgesic, and anti-diarrheal activity of the extracts of the leaves of Syzygium reticulatum. The leaves of the S. reticulatum were extracted with hexane and methanol by the maceration process successively, denoted as SRH and SRM. The extract’s phytochemical analyses found flavonoids, alkaloids, saponins, tannins, glycosides, and carbohydrates. The GC-MS analysis of the methanol extract identified 8 compounds. The major components were identified as Diisooctyl phthalate (96.903 %), 2-methyltetracosane (1.094 %), and others with fewer amounts. The result of the antioxidant study of the extracts as total phenolic content of SRH 8.92 ± 0.62 and SRM 340.107 ± 11.11 mg/g, respectively, total flavonoids content of SRH 7.85 ± 0.71 and SRM 105.16 ± 0.93 mg/g, IC50 values of the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay of SRM was 14.67 µg/mL, SRH was 41.26 µg/mL and ascorbic acid 14.08 µg/mL; the total antioxidant capacity of SRH was 21.33 ± 2.52 mg/g, whereas SRM was 353.71 ± 4.59 mg/g. The results of these studies suggest that methanol extract has a more potent antioxidant capacity than hexane extract. The methanol extract also exhibited higher toxicity (LC50 value 49.99 µg/mL) compared to hexane extract (LC50 value 370.348 µg/mL), as revealed by the Brine shrimp nauplii cytotoxic study. An analgesic study of the methanol extract found that a lower dose (250 mg/kg) has more significant analgesic activity than the higher dose and standard drug aspirin. At 90 min, the hot plate’s highest percentage of latency elongation was 110.32, while the acetic acid-induced writhing inhibition was 63.04 %. At a dose of 500 mg/kg, the methanol extract also shown good efficacy in mice suffering from castor oil and magnesium sulfate-induced diarrhea. The findings from the study of the leaves of the S. reticulatum exhibited that methanol extracts have significant antioxidant, cytotoxic, analgesic, and anti-diarrheal activity. HIGHLIGHTS The extract’s phytochemical analyses found flavonoids, alkaloids, saponins, tannins, glycosides, and carbohydrates The GC-MS analysis of the methanol extract identified 8 compounds. The major components were identified as Diisooctyl phthalate (96.903 %), 2-methyltetracosane (1.094 %) The antioxidant and cytotoxicity studies suggest that methanol extract has a more potent antioxidant and cytotoxic capacity than hexane extract The findings from in-vivo study of the leaves of the reticulatum also exhibited that methanol extracts have significant analgesic, and anti-diarrheal activity GRAPHICAL ABSTRACT

The purpose of the current study was to investigate the in vivo (analgesic, antidiarrheal, neurological, and cytotoxic) and in vitro (antioxidant, antimicrobial, thrombolytic and anthelmintic) activity of different fractions of methanolic extract of Momordica charantia. The antioxidant property was evaluated by DPPH radical scavenging assay, while antimicrobial activity was examined against three Gram (+) and one Gram (-) bacteria. Thrombolytic and anthelmintic activities were evaluated by using human blood serum and by recording paralysis and death time in earthworm, respectively. Cytotoxic activity was investigated in brine shrimp nauplii. Analgesic and antidiarrheal activities were evaluated in Swiss albino mice and neurological effect was evaluated by open field and Elevated plus-maze test (EPM). All fractions (n-hexane, carbon tetrachloride and chloroform) possess significant (p<0.05) cytotoxic activity. In case of thrombolytic activity, the highest concentration of methanolic extract produced a remarkable percentage of clot lysis (46.12%). The concentration of 1000 μg/ml produced a significant antibacterial activity against Gram positive Staphylococcus aureus and Gram negative E. coli. Aqueous fraction at a dose of 400 mg/kg body weight, was found to show promising analgesic activity. In case of antidiarrheal and anthelmintic activity, plant extract showed dose-dependent activity. Methanolic extract and its fractions failed to produce any neurological effect in both methods. The overall results of the study tend to suggest that the methanolic extract and its fractions have promising pharmacological activities.

The ethanolic extract of the dried fruit of Solanum nigrum Linn. (Family: Solanaceae) was assessed for its possible analgesic, antidiarrhoeal, antimicrobial, antioxidant and cytotoxic activity. Phytochemical screening of the ethanolic extract revealed the presence of carbohydrate, alkaloids, tannins, saponins, steroids, glycosides, and gums. In acetic acid induced writhing in mice, the ethanolic extract (250 and 500mg/kg) exhibited significant (p&lt;0.05 &amp; p&lt;0.01) inhibition of writhing reflex 51.39% and 66.67% respectively compared to standard diclofenac sodium. The fruit extract showed a significant (P&lt;0.01 and P&lt;0.001) antidiarrhoeal activity against castor oil induce diarrhoea in mice in which it decreased the frequency of defecation and increased the mean latent period at the dose of 250mg/kg and 500mg/kg body weight. The ethanolic extract showed moderate antibacterial activity against both gram-positive and gram-negative bacteria. In the qualitative antioxidant assay using DPPH (1, 1-diphenyl-2-picryl hydrazyl) the extract showed free radical scavenging properties. In the brine shrimp lethality test, the extract showed cytotoxicity significantly with LC50= 63.10μg/ml and LC90= 160μg/ml. All the results tend to justify the traditional uses of the plant and require further investigation to identify the chemicals responsible for these effects. Key words: Solanum nigrum Linn; analgesic activity; antidiarrheal activity; antimicrobial activity; antioxidant activity; cytotoxic activity. DOI: 10.3329/sjps.v3i1.6796S. J. Pharm. Sci. 3(1): 38-45

The methanolic crude extract of Lagerstroemia speciosa roots was investigated for its possible analgesic and anti-diarrhoeal activities in experimental animal model. Phytochemical screening of the methanolic extract of L. speciosa roots showed the presence of alkaloids, flavonoids, saponins, tannins and reducing sugar. In acute toxicity study, no mortality or toxic reaction was recorded in animal model after administration of the L. speciosa roots extract. Analgesic activity was evaluated by using acetic acid induced writhing inhibition method in Swiss albino mice. In peripheral method of anti-nociception, the methanolic crude extract of L. speciosa roots showed significant analgesic activity. At the dose of 200 and 400 mg/kg body weight, the extract produced 35.38% and 53.85% (P&lt;0.001) of writhing inhibition, respectively compared to standard diclofenac sodium (70.77% inhibition). The extract had also anti-diarrhoeal activity in castor oil induced method and inhibited the mean number of defecation by 32.75% (P&lt;0.01) and 51.72 % (P&lt;0.001) at the dose of 200 and 400 mg/kg body weight, respectively. The latent period for the extract treated group also increased significantly. DOI: http://dx.doi.org/10.3329/dujps.v13i1.21860 Dhaka Univ. J. Pharm. Sci. 13(1): 57-62, 2014 (June)

Plants are the priceless sources of bioactive natural compounds. Medicinal plants have been used since time immemorial in both developing and developed countries. Flemingia macrophylla (Willd.) is widely used as a hereditary medicines. The current study was designed to observe the analgesic, antidiarrheal and CNS depressant activities of methanol extract of the leaves of F. macrophylla (MEFM) and its petroleum ether (PEFM), dichloromethane (DFM), chloroform (CFM) and aqueous (AFM) fractions. The analgesic activities was assessed by acetic acid induced writhing method at doses 200- and 400- mg/kg body weight. The CNS-depressant effect was assayed by phenobarbitone sodium-induced sleeping time test. The anti-diarrheal activity of the extract was evaluated using castor oil-induced diarrhea in mice. The crude extract displayed significant peripheral analgesic activity at both test doses with 56.72- to 59.70-% inhibition of writhing responses, respectively. In CNS-depressant test, the extract revealed its activity in a dose dependent manner. In screening for antidiarrheal activity, the extract exhibited 20.83 and 41.67 % inhibition of defecation at 200- and 400-mg/kg bw, respectively whereas the standard loperamide (50 mg/kg bw) displayed 70.83 % inhibition of defecation. Bangladesh Pharmaceutical Journal 23(2): 141-145, 2020

Background. Elaeocarpus serratus L. (Family: Elaeocarpaceae) is a tropical fruit tree, traditionally used in the treatments of poisoning, diarrhea, arthritis, and other diseases. Objectives. The current study was performed to conduct the analgesic, antidiarrheal, and hypoglycemic activity of E. serratus in mice model using methanolic bark crude extract. Methods. To assess the peripheral and central analgesic activity, the acetic acid-induced writhing and tail immersion methods were respectively used. The castor-oil mediated antidiarrheal method was used to assess the antidiarrheal activity whereas, the tail tipping technique was conducted to determine the hypoglycemic activity of the plant extract. Results. In the peripheral analgesic assay, the methanolic bark crude extract of E. serratus significantly inhibits the number of writing 69.77% (200 mg/kg) and 73.26% (400 mg/kg) respectively (p&lt;0.05) which was strongly comparable with standard NSAID drug diclofenac sodium 75.58% (p&lt;0.05). Similarly, it shown a significant tail flicking response for 30 minutes, 60 minutes and 90 minutes of central analgesic activity assay. In antidiarrheal activity assay, the E. serratus substantially reduced the number of diarrheal feces 64.26% (200 mg/kg, p&lt;0.05) and 78.57% (400 mg/kg, p&lt;0.05) which was also comparable with the positive control loperamide. The hypoglycemic activity of E. serratus extract was not convincing. Conclusions. Our investigation demonstrated the significant analgesic and antidiarrheal activities of methanolic bark extract of E. serratus (200 and 400 mg/kg) in mice model.

Hevea brasiliensis, a plant belonging family Euphorbiaceae. In Brazil this plant is not only use for medicinal purpose but also for cosmetics purpose. The present study was aimed to study analgesic and antidiarrheal activity of methanol extract of Hevea brasiliensis. Analgesic activity was evaluated by acetic acid- induced writhing method and antidiarrheal by gastrointestinal motility method (charcoal meal test) in mice. Phytochemical evaluation was carried out by qualitative analysis. For analgesic evaluation, the extract (250 mg and 500 mg) showed significant activity compared to control diclofenac Na. On the other hand, for antidiarrheal activity the extract (250 mg and 500 mg) significantly reduce charcoal propulsion. The phytochemical evaluation showed significant presence of Alkaloids, Carbohydrates, Glycosides, Saponins, Phytosterols, Proteins and amino acids, Fats &amp; fixed oils. It’s concluded that the extract possesses both analgesic and antidiarrheal activity and containing wide range of phytochemicals.

BackgroundIn the current study in vitro antioxidant, thrombolytic and in vivo analgesic, anti-inflammatory, anti-diarrheal activities of methanol extract of Ixora cuneifolia leaves (MEIL) were investigated.MethodsThe antioxidant activity was determined by using DPPH free radical scavenging method at doses 25, 50, 100, 200, 400 and 800 μg/ml. Total phenolic content was determined at doses of 50, 100, 150, 200 and 250 μg/ml. Whereas thrombolytic activity was determined by clot lysis method. The analgesic activity was conducted in Swiss albino mice at doses of 250 and 500 mg/kg body weight by hot plate method and acetic acid induced writhing method. Carrageenan induced rat paw edema method was used to evaluate anti-inflammatory activity. For anti-diarrheal activity castor oil induced method was used at the same dose mentioned earlier.ResultsInitial phytochemical study revealed the presence of flavonoids, saponins, gums, terpenoids, steroids and reducing sugars. The extract possessed significant (p < 0.05-0.001) free radical scavenging properties (IC50 for DPPH scavenging activity is 57.64 64 μg/ml and also contained marked amount of phenols 46.44 ± 1.022 mg/gm (Gallic acid equivalents). In clot lysis activity the extract showed potent lysis capacity (48.41 %). In the hot plate method the extract produced a significant (p < 0.05) dose dependent reduction of thermally induced pain. The extract also showed significant (p < 0.001) inhibition of the total number of writhing produced in the acetic acid induced writhing method. Highest inhibition of writhing was 66 % at dose of 500 mg/kg body weight. In carrageenan induced paw edema test the extract at doses 250 and 500 mg/kg produced significant (p < 0.001) reduction in the volume of paw edema at the 6th hour following its administration. In case of anti-diarrheal activity a dose dependent reduction of fecal dropping was observed. Where the maximum reduction observed at the 2nd hour at 500 mg/kg dose (68.42 %) while the standard drug Loperamide produces 86.76 % reduction of fecal dropping.ConclusionsThe findings of our study suggested that the methanol extract of Ixora cuneifolia possess a significant antioxidant, thrombolytic, analgesic, anti-inflammatory and anti-diarrheal activity.

Background and Objectives: Microcos paniculata is widely distributed throughout the Bangladesh which belongs to the family Tiliaceae. The present study was considered to determine anti-diarrheal, analgesic, membrane stabilizing and thrombolytic activities with evaluation of phytochemical nature of methanolic extract of M. paniculata steam bark. Methods: Methanolic extract of this plant part was prepared at room temperature and the concentrated methanolic extracts used for the phytochemical screenings by using appropriate and proper procedure for the investigation of phytoconstituents. The extract was then used for investigation of in-vitro thrombolytic and membrane stabilizing activity. The in-vivo antidiarrheal and analgesic activities were done according to the method of castor oil and acetic acid induce writhing respectively. Results: Phytochemical study reveal the presence of flavonoids, diterpenes, alkaloids, saponin, tannin and phenols. During the estimation of anti-diarrheal activity plant extract showed most significant inhibition of 63.30% and 56.70% diarrhea at 400 mg/kg and 200 mg/kg, respectively. Analgesic test showed pain inhibition of 41.36% and 32.0% at 500 mg/kg and 250 mg/kg, respectively in contrast to standard drug (Ibuprofen) showed 46.64% inhibition of pain. The membrane stabilizing study showed that the inhibition of hemolysis increases respectively when the dose increased. Similar result was found in thrombolytic activity which means that when the dose increased, the thrombolytic activity of the plant extract increased. Conclusion: By considering this result, it can be concluded that M. paniculata may lead to find novel drugs for cardiovascular and diarrheal patient and also bring hope for patients suffering with diverse pain. Keywords: Microcos paniculata, phytochemical screening, thrombolytic, anti-diarrheal, analgesic, clot lysis.

Purpose: To assess the central nervous system (CNS) depressant, analgesic and antidiarrheal activities of the dried seed crude extracts of Dimocarpus longan Lour in rodents.Methods: Selected pharmacological effects of the ethanol (ENLS), petroleum ether (PELS), chloroform (CHLS) and ethyl acetate (EALS) extracts of D. longan fruit seeds were investigated. CNS depressant activity was evaluated by open field and hole cross tests; analgesic activity by acetic acid-induced writhing test and formalin-induced licking test; and anti-diarrheal activity was assessed in castor oil and magnesium-induced diarrhea rat model. The extracts were given orally in a rat model at doses of 200 and 300 mg/kg body weight. Normal saline served as control in all experiment. In CNS depressant test, diazepam (1 mg/kg) was used as reference drug while indomethacin (10 mg/kg) and loperamide(2 mg/kg) were used as standard drugs in analgesic and antidiarrheal tests, respectively.Results: In hole cross method, EALS showed the most effective depressant effect, viz, 1.17±0.17 for 200 mg/kg dose and 0.83±0.31 number of movements for 300 mg/kg dose after 120 min (p &lt; 0.01), whereas in the open field test, all the extracts exhibited significant (p &lt; 0.01) depressant effect in relation to positive control, diazepam. In acetic acid-induced pain test, PELS gave the lowest number of writhing (2.83±0.307) and the highest inhibition (88.45 %, 300 mg/kg dose) which was statistically significant. All the extracts also significantly (p &lt; 0.01) suppressed licking activity in both phases of the formalin-induced licking test, in contrast to indomethacin. In the antidiarrheal tests, diarrheal suppression was highest at 300 mg/kg dose for all the extracts, compared with loperamide in both castor oil and magnesium sulphate induced diarrhea model.Conclusion: The extracts of Dimocarpus longan tested demonstrated significant CNS depressant, analgesic and antidiarrheal activities in a rodent model.Keywords: Dimocarpus longan Lour, CNS depressant, Analgesic, Anti-diarrheal.

The current study was designed to evaluate the antidiarrheal and analgesic activities of the methanol extract of Bouea oppositifolia (Roxb.) Adelb. leaves through iv vivo studies in Swiss Albino mice. Oral administration of the extract at doses of 200- and 400-mg/kg of body weight demonstrated statistically significant (p &lt; 0.05) antidiarrheal activities in castor oil-induced diarrheal mice. The extracts and standard loperamide alleviated diarrhea by margins of 31.82-, 45.45- and 68.17%, respectively. Potential of the plant extract to effectuate analgesia was ascertained both centrally and peripherally. In tail immersion method for the determination of central analgesic activity, the plant extract at both doses enhanced pain tolerance to maximum extents of 192.76% and 221.09%, respectively, compared to standard morphine (419.57%) after 90 minutes of sample administration. Comparable levels of central analgesia were also observed for both doses of the plant extracts at 30 and 60 minutes of pharmacological intervention and the p value of less than 0.05 illustrated statistical significance of the activity. On a similar note, potent peripheral analgesic activities were observed for both doses of the extract as evident by 45.45- and 54.55% inhibitions of acetic acid induced writhing responses, respectively. Compared to the standard acetylsalicylic acid characterized by 78.18% inhibition, the peripheral analgesic activity of the plant was found to be statistically significant (p &lt; 0.05). The results of the study are indicative of the presence of potentially bioactive phytoconstituents with antidiarrheal and analgesic properties which might lead to newer drug candidates in future. Bangladesh Pharmaceutical Journal 23(2): 167-171, 2020

The present study was aimed for the evaluation of analgesic, anti-diarrheal and anti-depressant activities of methanol crude extract of Bridelia verrucosa leaves in Swiss albino mice. The analgesic activity was assessed by formalin-induced paw licking method, where the crude extract of B. verrucosa (400 mg/kg, b.w.) exhibited 54.55% reduction of licking response in mice as compared to 69.09% reduction exhibited by standard acetylsalicylic acid. In the castor oil-induced diarrhea in mice, the plant extract at the dose of 400 mg/kg, b.w., demonstrated 57.67% reduction of diarrheal feces in mice. The antidepressant activity of the plant extract was evaluated through phenobarbitone-Na induced sleeping time test. The methanol extract of B. verrucosa and its different Kupchan fractions were also subjected to screenings for membrane stabilizing and cytotoxic activities using acetylsalicylic acid and vincristine sulphate as standard, respectively. Bangladesh Pharmaceutical Journal 22(1): 50-55, 2019