ABSTRACT
The present study used in vitro and in silico techniques, as well as the metabolomics approach to char-acterise α-glucosidase inhibitors from different fractions of Clinacanthus nutans. C. nutans is a medicinal plant belonging to the Acanthaceae family, and is traditionally used to treat diabetes in Malaysia. n-Hexane, n-hexane: ethyl acetate (1:1, v/v), ethyl acetate, ethyl acetate: methanol (1:1, v/v), and methanol fractions were obtained via partitioning of the 80％ methanolic crude extract. The in vitro α-glucosidase inhibitory activity was analyzed using all the fractions collected, followed by profiling of the metabolites using liquid chromatography combined with mass spectrometry. The partial least square (PLS) statistical model was developed using the SIMCA P +14.0 software and the following four inhibitors were obtained:(1) 4,6,8-Megastigmatrien-3-one; (2) N-Isobutyl-2-nonen-6,8-diynamide; (3) 1′,2′-bis(acetyloxy)-3′,4′-didehydro-2′-hydro-β, ψ-carotene; and (4) 22-acetate-3-hydroxy-21-(6-methyl-2,4-octadienoate)-olean-12-en-28-oic acid. The in silico study performed via molecular docking with the crystal structure of yeast isomaltase (PDB code: 3A4A) involved a hydrogen bond and some hydrophobic interactions be-tween the inhibitors and protein. The residues that interacted include ASN259, HID295, LYS156, ARG335, and GLY209 with a hydrogen bond, while TRP15, TYR158, VAL232, HIE280, ALA292, PRO312, LEU313, VAL313, PHE314, ARG315, TYR316, VAL319, and TRP343 with other forms of bonding.
ABSTRACT
Clinacanthus nutans (C. nutans) leaf extracts have been widely used by cancer patients in Malaysia and local practiceclaims a cure to cancer. There were several studies done to determine the cytotoxicity potency of C. nutans extracts onvarious types of cells. However, there is still lacking on the knowledge regarding the combination effect of C. nutanswith anticancer drugs. Thus, the study was carried out to determine the cytotoxicity potency of C. nutans extracts andpaclitaxel (PTX) alone and, in combination on MDA-MB-231 cells. The cells were treated with 100% ethanol extract ofC. nutans (CNE) and water extract of C. nutans (CNA), PTX and combination of both extracts and PTX for 72 hours andthe cytotoxic activity was determined using SRB assay. Result showed that CNE had little cytotoxic activity, whereas CNAshowed no cytotoxic activity on MDA-MB-231 cells. For combination treatment of C. nutans extracts and PTX, only CNEshowed significant enhanced PTX-induced cytotoxicity (p < 0.05), meanwhile CNA inhibited PTX-induced cytotoxicitysignificantly (p < 0.05). As a conclusion, CNE was able to increase PTX potency to inhibit the viability of MDA-MB-231cells.
ABSTRACT
@#Traditionally, Clinacanthus nutans (CN) or locally named as ‘Belalai Gajah’ is one of the herbal plant claimed to beable to treat cancer. The aimd of this study are to extract, isolate and characterize the active anticancer compoundfrom CN and to determine the mode of cell death induced by the compound. Bioassay guided fractionation was done onthe CN extract by using column chromatography. The cytotoxicity activities of these fractions toward HeLA cells wereexamined by MTT assay. The nuclear morphology was examined by Hoechst 33258 staining and the cell cycle arrestwas evaluated by propium iodide staining using flow cytometry. The presence of active compound in the chosen fractionwas determined by Liquid Chromatography Mass Spectrometry (LCMS). Out of 16 fractions collected, Fraction 11(F11)showed the lowest IC50 value with 27 ± 2.6 µg/mL. The value of IC50 for F11 towards normal cell, NIH 3T3 cell and L929cell, were 70 ± 4.0 µg/mL and 45 ± 1.5 µg/mL respectively. These values were higher than tamoxifen, therefore indicatingthat tamoxifen is more toxic towards normal cells compared to F11. Nuclear morphology of HeLA cell displayed DNAfragmentation, nuclear condensation and formation of apoptotic bodies upon treatment with F11 for 24 hours. The cellcycle distribution of HeLA cell treated with F11 was arrested at G1 phase. The active compound identified to potentiallypossess the anticancer property is 19-Oxo-all-trans-retinoic acid. In conclusion, 19-Oxo-all-trans-retinoic acids fromF11 of the CN extract, is a potential anticancer agent for cervical cancer.
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@#Clinacanthus nutans (C. nutans), a well-known ethnopharmacological plant consumed for its medicinal purposes by Southeast Asian communities. C. nutans is said to possess antipyretic, inflammatory, antiedemic as well as analgesic properties and used traditionally in treating various skin ailments, Herpes infection, cancer and diabetes. The young leaves of this C. nutans are consumed in Malaysia for maintaining health. In this study, the proliferative activity of human gingival fibroblast cells (HGF-1, ATCC®CRL-2014™, USA) treated with the ethanol extract obtained from C. nutans leaves at three different concentrations (250, 125 and 62.5 µg/ml) was compared with the untreated cells using alamarBlue assay. The proliferative activity of HGF-1 using alamarBlue assay showed that the cells treated with 62.5 μg/ml of ethanolic extract of C. nutans leaves exhibited increased proliferation compared to the other groups and hence does not exhibit any cytotoxicity on HGF-1.
ABSTRACT
Clinacanthus nutans Lindau is known as snake grass belonging to the Acanthaceae family. This plant has diverse and potential medicinal uses in traditional herbal medicine for treating skin rashes, insects and snake bites, lesions caused by herpes simplex virus, diabetes, and gout in Malaysia, Indonesia, Thailand and China. Phytochemical investigations documented the varied contents of bioactive compounds from this plant namely flavonoids, glycosides, glycoglycerolipids, cerebrosides and monoacylmonogalatosylglycerol. The pharmacological experiment proved that various types of extracts and pure compounds from this species exhibited a broad range of biological properties such as anti-inflammatory, antiviral, antioxidant, and anti-diabetic activities. The findings of toxicity study showed that extracts from this plant did not show any toxicity thus it can be used as strong therapeutic agents for specific diseased conditions. However, further experiments on chemical components and their mode of action showing biological activities are required to elucidate the complete phytochemical profile and assess to confirm their suitability for future drugs. This review summarizes the medicinal uses, phytochemistry and pharmacology of this plant in order to explore its therapeutic potential and gaps necessitating for prospected research work.
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Objective: To evaluate the monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG) from Clinacanthus nutans (C. nutans) for their in vitro antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay. Methods: MGDG and DGDG were extracted with chloroform from C. nutans leaves. MGDG and DGDG were separated from chloroform crude extract using column chromatography, characterized by thin layer chromatography and quantified by high performance liquid chromatography. The anti HSV-1 and 2 activity against pre-treatment and post-treatment of the compounds was evaluated using plaque reduction assay. The cytotoxicity of the extract and the compounds on Vero cells were performed by MTT assay. Results: MGDG and DGDG obtained by column chromatography showed identical profiles as standard MGDG and standard DGDG using thin layer chromatography and high performance liquid chromatography. MGDG and DGDG from C. nutans showed 100% inhibition of HSV-1 replication at the post step of infection at noncytotoxic concentration with IC
ABSTRACT
Clinacanthus nutans Lindau is known as snake grass belonging to the Acanthaceae family. This plant has diverse and potential medicinal uses in traditional herbal medicine for treating skin rashes, insects and snake bites, lesions caused by herpes simplex virus, diabetes, and gout in Malaysia, Indonesia, Thailand and China. Phytochemical investigations documented the varied contents of bioactive compounds from this plant namely flavonoids, glycosides, glycoglycerolipids, cerebrosides and monoacylmonogalatosylglycerol. The pharmacological experiment proved that various types of extracts and pure compounds from this species exhibited a broad range of biological properties such as anti-inflammatory, antiviral, antioxidant, and anti-diabetic activities. The findings of toxicity study showed that extracts from this plant did not show any toxicity thus it can be used as strong therapeutic agents for specific diseased conditions. However, further experiments on chemical components and their mode of action showing biological activities are required to elucidate the complete phytochemical profile and assess to confirm their suitability for future drugs. This review summarizes the medicinal uses, phytochemistry and pharmacology of this plant in order to explore its therapeutic potential and gaps necessitating for prospected research work.
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Objective: To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities. Methods: Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted withn-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay. Results: Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) μg/mL, (44.50±2.66) μg/mL, (64.93±7.00) μg/mL, respectively where as those of C. siamensis were (60.00±11.61) μg/mL, (55.69±4.41) μg/mL, (37.39±5.85) μg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanolC. nutans leaves extracts were (72.62±12.60) μg/mL, (65.19±21.45) μg/mL, (65.13±2.22) μg/mL, respectively where as those of C. siamensis were (46.52±4.08) μg/mL, (49.63±2.59) μg/mL, (72.64±6.52) μg/mL, respectively. Conclusions: The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.
ABSTRACT
<p><b>OBJECTIVE</b>To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities.</p><p><b>METHODS</b>Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted with n-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay.</p><p><b>RESULTS</b>Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) µg/mL, (44.50±2.66) µg/mL, (64.93±7.00) µg/mL, respectively where as those of C. siamensis were (60.00±11.61) µg/mL, (55.69±4.41) µg/mL, (37.39±5.85) µg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanol C. nutans leaves extracts were (72.62±12.60) µg/mL, (65.19±21.45) µg/mL, (65.13±2.22) µg/mL, respectively where as those of C. siamensis were (46.52±4.08) µg/mL, (49.63±2.59) µg/mL, (72.64±6.52) µg/mL, respectively.</p><p><b>CONCLUSIONS</b>The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.</p>
Subject(s)
Humans , Acanthaceae , Chemistry , Genetics , Antiviral Agents , Chemistry , Pharmacology , Flowers , Chemistry , Cell Biology , Genetics , Herpesvirus 1, Human , Herpesvirus 2, Human , Phenotype , Plant Extracts , Chemistry , Pharmacology , Plant Leaves , Chemistry , Cell Biology , Genetics , Simplexvirus , Viral Plaque Assay , Virus ReplicationABSTRACT
The effect of crude extract of Clinacanthus nutans (CN) was studied to determine the antiviral activity against varicella-zoster virus (VZV) with three different treatments. Specifically the effects studied were that of CN extract on (I) cells before infection (pre-treatment); (II) virus infected cells (post-treatment); and (III) virus directly (inactivation assay). After treatment, the virus was detected by methods of DNA hybridization and plaque reduction assay. It was shown that CN had an effect on VZV depending on concentration and methods of treatment. Via DNA hybridization, the ID50 (50% inhibitory dose) of pre-treatment, post-treatment, and inactivation assay was by weight per volume dilution 1:2,000, 1:6,000 and > 1:18,000, respectively; by plaque reduction assay, they were 1:2,000, 1:4,800 and 1:9,600, respectively. From the present findings, based on the result of inactivation assay, it was recognized that the in vitro antiviral activity of CN might be a direct interaction of the extract with the virus.