Clinacanthus Nutans Research Articles

English

Clinacanthus nutans is an economically important medicinal plant that can be found grown in many countries in the Asian region. Useful medicine properties such as anti-cancer, anti-bacteria, and anti-viral, backed by its high content of phytochemical compounds such vitexin, isovitexin, stigmasterol and lupeol has increased the demand for C. nutans in the market. Extensive work had been carried out on its content and pharmacological activity using fresh samples but limited studies for in vitro cultures of C. nutans. No genome size estimation has been done for C. nutans. Objective of this study was to analyse nuclear DNA content of C. nutans using flow cytometry. Preparation of different nuclei isolation buffer and stoichiometric DNA staining using propidium iodide was carried out. The genome size of C. nutans was estimated using Glycine max cv. Polanka as internal standard and its genome size was compared with in vitro plantlets of C. nutans. Flow cytometry analysis revealed that nuclear 2C DNA of C. nutans content is estimated at 1.75 ± 0.006 pg. Coefficient of variation in flow cytometric analysis was within the limit of 5% implying that the results were reliable with the Tris.MgCl2 being the best nuclei isolation buffer. No significant difference was observed from field grown and in vitro C. nutans. This finding will assist further in genome size evolution analysis of Clinacanthus spp. and to determine polyploids for increased active compounds and biomass. Key word: Flow cytometry; Nuclear DNA content, Clinacanthus nutans.

Cytotoxic and Antiviral Activities of Clinacanthus nutans Crude Extract

Clinacanthus nutans, also known as snake grass, is a local medicinal plant that displays antimicrobial, antiviral, and anticancer immunomodulatory activities. Whole dried plants were macerated with methanol for crude extract preparation. The focus of this study was to assess the cytotoxicity and antiviral activities of the C. nutans crude extract. Cytotoxicity screening against Vero cells using MTT assay showed that the CC50 value of the crude extract of C. nutans was 1.625 mg/mL. Plaque reduction assays were carried out to evaluate the antiviral activity of the extract against herpes simplex virus type 1 (HSV-1). These included post-treatment, pre-treatment, and virucidal assays. The selective indices (SI = CC50 / EC50) of the extract in post-treatment, pre-treatment, and virucidal assays were 11, 6, and 3, respectively. Our results indicate that C. nutans has a promising potential to be explored as a source of potent antiviral products.

A Narrative Review on the Phytochemistry, Pharmacology and Therapeutic Potentials of Clinacanthus nutans (Burm. f.) Lindau Leaves as an Alternative Source of Future Medicine.

The application of natural products and supplements has expanded tremendously over the past few decades. Clinacanthus nutans (C. nutans), which is affiliated to the Acanthaceae family, has recently caught the interest of researchers from the countries of subtropical Asia due to its medicinal uses in alternative treatment for skin infection conditions due to insect bites, microorganism infections and cancer, as well as for health well-being. A number of bioactive compounds from this plant’s extract, namely phenolic compounds, sulphur containing compounds, sulphur containing glycosides compounds, terpens-tripenoids, terpens-phytosterols and chlorophyll-related compounds possess high antioxidant activities. This literature search yielded about one hundred articles which were then further documented, including the valuable data and findings obtained from all accessible electronic searches and library databases. The promising pharmacological activities from C. nutans leaves extract, including antioxidant, anti-cancer, anti-viral, anti-bacterial, anti-fungal, anti-venom, analgesic and anti-nociceptive properties were meticulously dissected. Moreover, the authors also discuss a few of the pharmacological aspect of C. nutans leaves extracts against anti-hyperlipidemia, vasorelaxation and renoprotective activities, which are seldom available from the previously discussed review papers. From the aspect of toxicological studies, controversial findings have been reported in both in-vitro and in-vivo experiments. Thus, further investigations on their phytochemical compounds and their mode of action showing pharmacological activities are required to fully grasp both traditional usage and their suitability for future drugs development. Data related to therapeutic activity and the constituents of C. nutans leaves were searched by using the search engines Google scholar, PubMed, Scopus and Science Direct, and accepting literature reported between 2010 to present. On the whole, this review paper compiles all the available contemporary data from this subtropical herb on its phytochemistry and pharmacological activities with a view towards garnering further interest in exploring its use in cardiovascular and renal diseases.

Chemical characterization, antiproliferative and antifungal activities of Clinacanthus nutans

Clinacanthus nutans Lindau (Family: Acanthaceae) is a medicinal herb widely distributed in the tropic and subtropic areas of Asia. C. nutans is traditionally consumed as vegetable or herbal tea, as well as a folk medicine for anticancer and antifungal activities. However, to date, chemical constituent responsible for observed health beneficial effects of this medicinal plant is not clear. In the current study, 32 compounds (1−32), including three new megastigmanes (1–3) were isolated from the aerial parts of C. nutans. Their structures were elucidated on the basis of comprehensive NMR, MS, and CD spectroscopic data analysis, as well as chemical hydrolysis. Among the isolates, cycloartane triterpenoids (9, 10, and 12) displayed moderate anti-proliferative effects against HepG2 cell growth with IC50 values ranging from 9.12 to 19.89 μM. Data obtained from flow cytometry analysis and western blotting assays revealed that compounds 9 and 12 induced apoptosis of HepG2 cells by modulating the expression of proteins associated to mitochondrial-mediated apoptotic pathway. Furthermore, megastigmanes 1, 2, 7, and 8 enhanced the anti-Candida albicans activity of amphotericin B (AmB), supporting the synergistic effects between megastigmanes and AmB. This is the first report of anticancer and antifungal potential of cycloartane triterpenoids and megastigmanes in C. nutans, which shed useful insights on the relationship between C. nutans's chemical constituent and its beneficial effects to health. Findings from this study support further development of this medicinal plant for potential pharmaceutical applications.

Study of antibacterial activity of Streptomyces sp. SS473 isolated from plants against ESBL-producing Escherichia coli

The broad spectrum β-lactamase-producing E. coli(ESBL) is a dangerous bacterial pathogen in humans due to its resistance to many antibiotics. This is especially serious in the context of a limited number of new antibiotics for treating bacterial infections. This leads to a global public health threat and places an urgent need for new antibiotics. In this study, the authors investigated the antibacterial properties of an actinomyces strain isolated from the plant Clinacanthus nutans against the ESBL-producing E. coli strains. These actinomyces strains were designated as SS473. Moreover, SS473 showed a broad spectrum of antibacterial activity on several clinically isolated pathogenic bacteria. Culture media have different effects on the antibacterial activity of SS473. In stability tests, the antibacterial activity of strain SS473 remained at a temperature up to 80oC but was lost at pH 3 and 13. By contrast, the antibacterial activity was not affected by UV and protease treatments. Based on the results of morphological identification with specific media for Streptomyces and molecular identification on 16S rRNA gene, strain SS473 was suggested to belong to the Streptomyces genus and was named Streptomycessp. SS473. The results in this study will pave the way for the following research on the identification of secondary metabolites having antibacterial activity and their biosynthetic pathways in Streptomyces sp. SS473 in the future

Osteogenic Activity of Lupeol Isolated from Clinacanthus nutans Lindau: Activity and Mode of Action

Clinacanthus nutans Lindau has been traditionally used for healing of bone fragility, but the mechanism of actions has not been clarified yet. In this study, the bone regeneration activity of lupeol derived from C. nutans was assessed using an in vitro model of osteoblast cells MC3T3-E1. The finding revealed that the compound was not significantly toxic to osteoblast cells at concentration of ≤40 μg/mL. Lupeol demonstrated the osteogenic activity through enhancement of alkaline phosphatase (ALP) of osteoblast cells up to 31.2%, 21%, and 12% at concentrations of 5, 10, and 20 µg/mL, respectively ( p < 0.05). Besides, the mineralization activity was increased up to 170, 230, 185, and 117% at concentration of 5, 10, 20, and 40 μg/mL, respectively ( p < 0.05). The marker genes related to osteoblast differentiation evaluated on the expression level in the presence of lupeol, including collagen I (col 1), osteopontin (opn), osterix (osx), and runx2, showed upregulated expression in all the test genes ( p < 0.05). The Western blot analysis demonstrated a clear effect of lupeol on expression of p38/p-p38, and ERK/p-ERK proteins involved in the MAPK signaling pathway. Thus, lupeol isolated from C. nutans exhibited the osteogenic activity by enhancing expression of important markers of osteogenesis, as well as affected the MAPK signaling pathway relating to osteoblast differentiation. This is the first report on the detailed mechanism of action of lupeol on bone regeneration and also explains for the traditional use of this medicinal plant for bone healing.

Expression Pattern of Genes in Condyloma Acuminata Treated with Clinacanthus nutans Lindau Cream versus Podophyllin.

Clinacanthus nutans Lindua (C. nutans), a strong antiviral traditional medicine, can be used to treat condyloma acuminata (CA) caused by the human papillomavirus (HPV). However, its molecular mechanism for CA elimination is unknown. Herein, we conducted a randomized clinical trial to evaluate the effectiveness of C. nutans and its molecular mechanism compared with podophyllin, the gold standard treatment. Using a randomized block design, six patients were treated with C. nutans and podophyllin for four weeks. Efficacy of drugs was assessed by size reduction of the warts and HPV viral load quantification using droplet digital PCR. The gene expression profiling of CA was analyzed using NanoString Technology. After the podophyllin and C. nutans treatments, CA lesion sizes were reduced to 97.0% and 84.4% clearance, and the HPV viral loads were reduced by 74.0% and 46.6%, respectively. The gene expression pattern of immune profiling showed that 23 genes (i.e., HLA-DPB, CCL3, CXCL2, CXCR2, and OSM) were significantly differentially expressed by podophyllin, whereas 2 genes (IFNL1 and IRF2) were remarkably expressed by C. nutans. In inflammatory profiling, 108 genes (i.e., CXCL2, IL8, and STAT3) were highly expressed by podophyllin, but none of genes were observed to change expression by C. nutans. These results suggested that podophyllin may reduce the HPV infection through a mechanism related to proinflammatory response. In addition, C. nutans was found to suppress the HPV infection through mechanism related to the activation of immune response. This study shows novel therapeutic mechanisms of podophyllin and C. nutans. It is suggested that C. nutans might be used as an alternative treatment for CA treatment.

Evaluation of Cytotoxic Mechanisms of Clinacanthus Nutans Extracts in Cancer Cells

Background: The preparations of commercially available Clinacanthus nutans (Burm.F) Lindau (Acanthaceae) (CN) leaf are gaining attention as an alternative cancer treatment, particularly in South East Asia. Multiple studies have suggested that CN has potential anticancer activities; however, the mechanism of these activities has remained elusive. Objective: This study evaluated the cytotoxic mechanisms of CN extracts in cancer cells. Methods: CN extracts were prepared from either fresh or dried leaves, using different solvents. Cytotoxicity of CN extracts was tested on the A549 (lung cancer), HeLa (cervical cancer), MCF-7 (breast cancer) and MDA-MB-231 (breast cancer) cell lines using the MTT assay. Flow cytometry was used to assess changes in the cell cycle profile, while Western blotting was used to examine microtubule stability. Finally, the mode of cell death was investigated using the Annexin V-FITC Apoptosis Detection Kit. Results: Aqueous Fresh (AQF) extract was prepared to simulate the ethnomedicinal use of CN, and reduce cell viability of MCF-7 cells with IC50 = 1.71 mg/mL. Some CN extracts have the ability to inhibit the proliferation of four different cancer cell lines after a 24 hour treatment. Annexin V assay results showed that acetone extracts of CN induced increments in the percentage of apoptotic cell death. However, flow cytometry results showed that the cancer cell cycle profile was not affected. Similarly, immunoblotting results also indicate that microtubule dynamics in MCF-7 cells were not altered. However, the aqueous extract, prepared to simulate the current ethnomedicinal use of CN leaves in cancer treatment, did not significantly inhibit cancer cell proliferation with an IC50 = 1.71 mg/mL. Conclusions: This study was the first to show that microtubules in cancer cells remain dynamic after treatment with CN extracts, effectively ruling interference of microtubule dynamics as the mode of cell death. AMD extract showed the highest effects of MCF-7 cell proliferation.

Immunomodulatory potential of Clinacanthus nutans extracts in the co-culture of triple-negative breast cancer cells, MDA-MB-231, and THP-1 macrophages.

Triple-negative breast cancer is the main type of breast carcinoma that causes mortality among women because of the limited treatment options and high recurrence. Chronic inflammation has been linked with the tumor microenvironment (TME) in breast cancer progression. Clinacanthus nutans (CN) has gained much attention because of its anticancer properties, but its mechanism remains unclear. We aimed to study the qualitative phytochemical content and elucidate the cytotoxicity effects of CN on human triple-negative breast cancer (TNBC), MDA-MB-231 and human macrophage-like cells such as THP-1 by using sulforhodamine B (SRB) assay. As highly metastatic cells, MDA-MB-231 cells can migrate to the distal position, the effect of CN on migration were also elucidated using the scratch assay. The CN effects on ameliorating chronic inflammation in TME were studied following the co-culture of MDA-MB-231/THP-1 macrophages. The cytokine expression levels of IL-6, IL-1β and tumor necrosis factor-alpha (TNF-α) were determined using ELISA assays. The results showed that both ethanolic and aqueous CN extracts contained alkaloid, phenol and tannin, flavonoid, terpenoid, glycoside and steroid. However, saponin was only found in the aqueous extract of CN. CN was not cytotoxic to both MDA-MB-231 and THP-1 cells. The ability of MDA-MB-231 to migrate was also not halted by CN treatment. However, CN ethanol extract decreased IL-6 at 25 μg/mL (p = 0.02) and 100 μg/mL (p = 0.03) but CN aqueous extract increased IL-6 expression at 50 μg/mL (p = 0.08) and 100 μg/mL (p = 0.02). IL-1β showed decreased expression after treated with CN ethanol and CN aqueous both at 25 μg/mL (p = 0.03). TNF-α were significantly decreased after CN ethanol treatment at concentration 25- (p = 0.001), 50- (p = 0.000) and 100 μg/mL (p = 0.000). CN aqueous extract slightly inhibited TNF-α at all 25-50- and 100 μg/mL (p = 0.001, p = 0.000, p = 0.000, respectively). Overall, CN acts by ameliorating the pro-inflammatory condition in the TME and may be a potential strategy for its anticancer mechanism on highly metastatic breast cancer condition. The major pathways that link both cancer and inflammation were NF-κB and STATs thus further study on the upstream and downstream pathways is needed to fully understand the mechanism of CN extracts in cooling the inflamed TME in breast cancer.

Mode-Dependent Antiviral Activity of Medicinal Plant Extracts against the Mosquito-Borne Chikungunya Virus.

The lack of specific treatment for chikungunya fever makes the need for anti-chikungunya virus agents more crucial. This study was conducted to evaluate 132 extracts obtained by sequential solvent extraction from 21 medicinal plants for cytopathic effect inhibitory activity using virus-infected Vero cells in two different sample introduction modes. Among the extracts, 42 extracts (31.8%) from 12 plants in the concurrent mode and three extracts (2.3%) from a plant in the non-concurrent mode displayed strong cytopathic effect inhibitory activity (cell viability ≥70%). Viral load quantification analysis unveiled that the extracts of Clinacanthus nutans (chloroform, ethyl acetate, and ethanol), Hydrocotyle sibthorpioides (ethanol), and Ocimum americanum (ethanol and methanol) hindered the release of viral progeny from the infected cells while the extracts of Ficus deltoidea (ethanol), Gynura bicolor (water), H. sibthorpioides (water), and O. americanum (chloroform and ethyl acetate) blocked the entry of virus into the cells. The extracts of Diodella sarmentosa (ethyl acetate), Diplazium esculentum (chloroform, ethyl acetate, and ethanol), and G. bicolor (ethanol) possessed virucidal effect and caused 5.41-log to 6.63-log reductions of viral load compared to the virus control. The results indicate that these medicinal plants are potential sources of anti-chikungunya virus agents that have varied modes of action.

Volatile Profiling Aided in the Isolation of Anti-Proliferative Lupeol from the Roots of Clinacanthus nutans (Burm. f.) Lindau

Isolation of anti-proliferative compounds from plants is always hindered by the complexities of the plant’s nature and tedious processes. Clinacanthus nutans (Burm. f.) Lindau is a medicinal plant with reported anti-proliferative activities. Our study aimed to isolate potential anti-proliferative compounds present in C. nutans plant. To start with, for our study, we came up with a strategy by first profiling the volatile compounds present in the leaf, stem and root of C. nutans using GC-MS. Comparing the plant’s volatile profiles greatly narrowed down our target of study. We decided to start with the isolation and characterization of a pentacyclic terpenoid, i.e., lupeol from the roots of C. nutans, as this compound was found to present abundantly in the roots compared to the leaf or stem. We developed a simple maceration and re-crystallization method, without the necessity to go through the fractionation or column chromatography for the isolation of lupeol. Characterizations of the isolated compound identified the compound as lupeol. The anti-proliferative activity of the isolated lupeol was further investigated against the MCF-7 cell line, which showed comparable anti-proliferative activity with the authentic lupeol and camptothecin. Our strategy to profile every part of the plant first, followed by selection of the most suitable plant part and targeted compound proved useful for further isolation and characterization bioactive compound from C. nutans.

Uji Aktivitas Antinosiseptif Kombinasi Ekstrak Daun Dandang Gendis [Clinacanthus nutans (Burn F) Lindau] Dan Daun Bakung (Crinum asiaticum L.) secara In Vivo

The development of new and effective pain medications that are natural isexpected to have the benefits of treatment. The purpose of this study was todetermine the antinociceptive effectiveness of a combination of 75% ethanolextract from dandang gendis leaves and 96% ethanol fraction of bakungleaf. In this study was used chemical method induction (Sigmund's method)in 25 male mice which were divided into 5 treatment groups, namelytreatment group I as negative control was given CMC 0.5%, treatmentgroup II as positive control was given acetosal 100 mg / kg BW, treatmentgroups III, IV and V were given combinations extract. Each group was giventreatment orally, thirty minutes then induced with 0.6% acetic acidintraperitonially, after which it was observed and counted for the amount ofstretching for sixty minutes. Data were analyzed by one-way ANOVA test.The results of this study showed that all treatment groups had a significantdifference (p <0.05), the combination of 75% ethanol extract dandanggendis leaf and 96% ethanol fraction of bakung leaf with a dose of 100 mg/ kg BW had good effectiveness with a percent effectiveness value of 92.50%.The results of this study indicate that the combination of 75% ethanolextract from dandang gendis leaf (Clinacanthus nutans (Burn F) Lindau)and 96% ethanol fraction from bakung leaf (Crinum asiaticum) with thesame concentration ratio, have relatively higher antinociceptive of theactivity of each extract, in other words both of them work together.

EFFECTS OF LIGHT ILLUMINATION AND SUBCULTURE FREQUENCY ON BIOMASS PRODUCTION IN CELL SUSPENSION CULTURES OF Clinacanthus nutans

Clinacanthus nutans (Burm. f.) Lindau is a medicinal plant rich in polyphenols and possesses multiple pharmacological properties. It is widely used as a side treatment for cancer and to prevent oxidative stress-related diseases. The present study aims to investigate the effects of light illumination and subculture frequency on the growth and proliferation of cells in the suspension culture of C. nutans. Friable callus induced from leaf explants C. nutans were cultured in liquid Murashige & Skoog medium supplemented with 0.25 mg/L 2, 4-dichlorophenoxyacetic acid, and 0.25 mg/L 6-benzylaminopurine to initiate the cell suspension culture. The suspension cultures were incubated under different conditions (16 hr photoperiod and continuous darkness). Subcultures were conducted every two weeks for ten subculture cycles. Results revealed that higher cell biomass was obtained under the exposure of 16 hr photoperiod with the highest biomass production at the fifth subculture cycle (growth index of 11.8) exhibiting stable growth from the eighth to the tenth subculture cycle. The suspension cells showed stable growth rates up to the 10th subculture cycle producing small aggregates and single cells. The present study indicated that light illumination significantly impacts the growth of suspension cultures for C. nutans in the production of valuable secondary metabolites.

Multi-Platform Metabolomics Analyses Revealed the Complexity of Serum Metabolites in LPS-Induced Neuroinflammed Rats Treated with Clinacanthus nutans Aqueous Extract.

The use of metabolomics as a comprehensive tool in the analysis of metabolic profiles in disease progression and therapeutic intervention is rapidly advancing. Yet, a single analytical platform could not be applied to cover the entire spectrum of a biological sample’s metabolome. In the present paper, multi-platform metabolomics approaches were explored to determine the diverse rat sera metabolites extracted from intracerebroventricular lipopolysaccharides (LPS)-induced neuroinflammed rats treated with oral therapeutic interventions of positive drug (dextromethorphan, 5 mg/kg BW); with Clinacanthus nutans (CN) aqueous extract (CNE, 500 mg/kg BW); and with phosphate buffer saline (PBS) as the control group for 14 days. Analyzed by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS) techniques, this study depicted the potential of metabolites associated with neuroinflammation and verified by MetDisease. The key observations in the perturbed metabolic pathways that showed ameliorative effects were linked to the class of amino acid and peptide metabolism involving valine, leucine, and isoleucine biosynthesis; phenylalanine, tyrosine, and tryptophan biosynthesis; and phenylalanine metabolism. Lipid metabolism of arachidonic acid metabolism, glycerophospholipid metabolism, terpenoid backbone biosynthesis, and glycosphingolipid metabolism were also affected. Current findings suggested that the putative biomarkers, especially lysophosphatidic acid (LPA) and 5-diphosphomevalonic acid from glycerophospholipid and squalene/terpenoid and cholesterol biosynthesis, respectively, showed the ameliorative effects of the drug and CN treatments by controlling cell differentiation and proliferation. Our study proved that the complex and dynamic sera profiling affected during the CN treatment was greatly influenced by the analytical platform selection as integration between the two data yielded a more holistic summary of the metabolite pattern changes. Hence, an evidence-based herb, such as CN, can be used for novel diagnostic tools in the quest for ethnopharmacological studies.

Recent Advancement in Anticancer Activity of Clinacanthus nutans (Burm. f.) Lindau.

Clinacanthus nutans is a traditional medicinal herb that is applied for the therapy of snake bites, skin infection, herpes infection, burns, scalds, dysentery, and diabetes. Clinacanthus nutans is also used to treat several cancers, including breast, cervical, colon, gastric, head and neck, liver, lung, pancreatic, and skin cancers, as well as lymphoma and leukemia; however, the underlying mechanisms of its anticancer activity remained undetermined. We searched PubMed and Google with key words “Clinacanthus nutans and cancer” and collected recent papers of Clinacanthus nutans with anticancer activity. We focused on the preparation, effects, and action mechanisms of Clinacanthus nutans extracts on various types of cancers. We hope that this mini review can help update our knowledge about active components, effects, and molecular mechanisms of extracts from this promising herb Clinacanthus nutans for ongoing studies and speed up its clinical application in the future.

Bibliographic analysis of Clinacanthus nutans papers in Scopus database (2000–2019)

Background: There has been an increasing trend in Clinacanthus nutans’ research, an important medicinal herb of Malaysia and Thailand, well known as an anti-viral, anti-cancer, and insect bite treatment. This study examines the trend in Clinacanthus nutans’ research from 2000 to 2019 and compares the contribution of research on this topic from different institutions and authors. Methods: Publications from the Scopus database were retrieved using keywords and identify top ten institutions/universities, list of prominent authors, top ten journals that published research, top five influential articles, top fifty cited papers, and global distribution of publications on Clinacanthus nutans. Microsoft Excel 2016, Wordcloud, SPSS version 26, and GunnMap 2 were used to analyse indicators. A total of 167 articles were identified from the Scopus database. All research publications were screened initially. Five articles (n=5) were removed due to the unavailability of the full-text version of the article. 162 articles were included in the final study. Results: Universiti Putra Malaysia and the Journal of Ethnopharmacology published the highest number of articles on Clinacanthus nutans. Herpes, antioxidant, phenolic, flavonoids, cancer, antimicrobial were common keywords identified using a word cloud. Over the past 20 years, the literature on Clinacanthus nutans has continuously grown, with the rate increasing after 2012. Conclusion: The prominent research on Clinacanthus nutans was based upon their identified and isolated bioactive constituents, and there is a need for more research on clinical trials.

Biogenic Silver Nanoparticles of Clinacanthus nutans as Antioxidant with Antimicrobial and Cytotoxic Effects.

Silver nanoparticles (AgNPs) previously synthesised using leaf (AgNP-L) and stem (AgNP-S) extracts of Clinacanthus nutans (C. nutans) were tested to evaluate antimicrobial, antioxidant, and cytotoxicity activities. The AgNPs showed good inhibition against bacteria, but not fungi. The inhibition results showed the highest activity against Staphylococcus aureus (S. aureus) with 11.35 mm (AgNP-L) and 11.52 mm (AgNP-S), while the lowest inhibition was against Escherichia coli (E. coli) with 9.22 mm (AgNP-L) and 9.25 mm (AgNP-S) in the disc diffusion method. The same trend of results was noted in the well diffusion method. The IC50 of AgNP-L and AgNP-S in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays was 417.05 μg/mL and 434.60 μg/mL, as well as 304.31 μg/mL and 326.83 μg/mL, respectively. Ferric reducing power (FRAP) assay showed that AgNP-L [872.389 μmol/L Fe(II)] and AgNP-S [612.770 μmol/L Fe(II)] exhibited significantly (p < 0.05) greater antioxidant activities than leaf extract (CNL) [152.260 μmol/L Fe(II)] and stem extract (CNS) [110.445 μmol/L Fe(II)] of C. nutans. The AgNPs were also proven to possess cytotoxic effects on the breast (MCF-7), cervical (HeLa), and colon (HT-29) cancer cells in a dose-dependent manner. AgNP-S and AgNP-L showed significantly (p < 0.05) higher cytotoxicity against MCF-7 (117.43 μg/mL) and HT-29 (78.47 μg/mL), respectively. In conclusion, the biosynthesised AgNPs from aqueous extract leaves and stem of C. nutans have demonstrated promising potential towards antioxidant, antimicrobial, and cytotoxicity activities.

Impact of commercial cellulase enzyme on the quality of Clinacanthus nutans extracts

The use of enzymes such as cellulase and pectinase to aid in the fruit and vegetable juice extraction is a common practice. However, this practice is very limited in the production of juice or extract from green leaves. This study was carried out to evaluate the impact of commercial cellulase enzyme on the volume yield recovery, total chlorophyll and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity of the Clinacanthus nutans leaves and stem. Leaves extract showed higher volume yield recovery, total chlorophyll content and DPPH scavenging activities (0.72±0.02 mL, 50.10±3.96 mg/L and 70% respectively) compared to the stem extract (0.60 ± 0.1 mL, 39.70 ± 4.73 mg/L and 25% respectively). It was also found that the cellulase enzyme concentrations were proportional to the volume yield recovery, and DPPH scavenging activity of the C. nutans extract over time. However, there was no significant effect of enzyme concentration on the total chlorophyll content. The results obtained can be considered very satisfactory and cellulase enzyme can be considered to be used in the aqueous extraction of C. nutans to aid the extraction process in order to increase the volume yield recovery, total chlorophyll content and DPPH scavenging activities.

Effects of Clinacanthus nutans Extracts on Cytokine Secretion in PMA-Induced U937 Macrophage Cells

Background and objectives: Clinacanthus nutans (Burm f.) Lindau (C. nutans) is a well-known traditional medicine in South East Asia and consists of abundant phytomedicinals properties. This study aimed to investigate the effects of C. nutans ethanol and aqueous extracts on interleukin-4 (IL-4) and interleukin-13 (IL-13) cytokines secretion in phorbol 12-myristate 13-acetate phorbol-12-myristate-13-acetate (PMA)-induced U937 macrophages. Methods: Sequential ultrasonic-assisted extraction was carried out using ethanol (ETOH) and water, by applying 1:10 ratio of leaves powder to the solvent volume. U937 cells were incubated with 25 nM PMA for 72 h to induce macrophage differentiation. The macrophage differentiation was assessed based on the cell morphological changes, cell viability and, CD14 and CD11b expression by using flow cytometry. The macrophages were incubated with both ETOH and aqueous extracts at 0.25, 0.5, 1.0, 2.0, 4.0 and 8.0 mg/mL concentration for 48 h. The viability of the extract-treated cells was assessed using PrestoBlue cell viability assay and the IL-4 and IL-13 secretions were assessed by using Enzyme-Linked Immunosorbent Assay (ELISA). Results: PMA stimulation caused morphological changes of U937 cells from round-shaped, non-adherent to larger irregular-shaped, adherent cells, and a reduction of cells viability to 87%. CD14 expression was down-regulated from 7% to 4.5% upon PMA stimulation. CD11b expression was up-regulated from 16% in untreated cells to 38% in PMA-treated cells. ELISA results showed that 1 mg/mL of ETOH and AQ extracts stimulated 1200 and 1800 pg/mL IL-4 secretions, respectively. However, both extracts caused minimal IL-13 secretion. Conclusion: Clinacanthus nutans aqueous extracts stimulated IL-4 production higher than ETOH extract in PMA-induced U937 macrophages.

The effect of different growing media on growth performance of Clinacanthus nutans

The effect of different growing media on the growth performance was tested for Clinacanthus nutans (Sabah snake grass). Five different growth media including T1: soil, T2: soil: coco peat (2:1), T3: soil: peat moss (2:1), T4: soil: coco peat (3:1) and T5: soil: peat moss (3:1) were used for growing Clinacanthus nutans. Height of the plants (cm), number of leaves, number of side branches, length of side branches (cm), root length (cm) and root weight (g) were determined. The chemical properties of each growth medium, including pH, nitrogen, phosphorus and potassium were also determined. The aimed of this study was to measure the growth rate of Clinacanthus nutans affected by the different growing media. The findings of this study showed the height of the plants (cm), number of leaves, length of side branches (cm), root length (cm) and root weight (g) were affected significantly when the plants were grown in soil mix with peat moss in T3. The finding of this study showed that the increased of nutrient in the growth media can be useful in the growth performance of the Clinacanthus nutans.