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Apoptotic activities of thymoquinone, an active ingredient of black seed (Nigella sativa), in cervical cancer cell lines

Ichwan SJ, Al-Ani IM, Bilal HG, Suriyah WH, Taher M, Ikeda MA

Chin J Physiol, 2014 Oct 31;57(5):249-55.
PMID: 25241984 DOI: 10.4077/CJP.2014.BAB190

Thymoquinone (TQ) is the main constituent of black seed (Nigella sativa, spp) essential oil which shows promising in vitro and in vivo anti-neoplastic activities in different tumor cell lines. However, to date there are only a few reports regarding the apoptotic effects of TQ on cervical cancer cells. Here, we report that TQ stimulated distinct apoptotic pathways in two human cervical cell lines, Siha and C33A. TQ markedly induced apoptosis as demonstrated by cell cycle analysis in both cell lines. Moreover, quantitative PCR revealed that TQ induced apoptosis in Siha cells through p53-dependent pathway as shown by elevated level of p53-mediated apoptosis target genes, whereas apoptosis in C33A cells was mainly associated with the activation of caspase-3. These results support previous findings on TQ as a potential therapeutic agent for human cervical cancer.
The phytochemical content and antimicrobial activities of Malaysian Calophyllum canum (stem bark)

Alkhamaiseh SI, Taher M, Ahmad F, Qaralleh H, Althunibat OY, Susanti D, et al.

Pak J Pharm Sci, 2012 Jul;25(3):555-63.
PMID: 22713941

Recently there was huge increase in using of 'herbal products'. These can be defined as plants, parts of plants or extracts from plants that are used for curing disease. However, Calophyllum species is a tropical plant and it has been used in traditional medicine, the limitation in safety and effectiveness information could lead to serious health problems. Providing information for communities by evaluating the phytochemical contents, antioxidant, antimicrobial and cytotoxic activities will improve the therapeutic values. Three main Calophyllum canum fractions (none - high polar) were tested to find out the phenolic, flavonoid, flavonol content, DPPH radical scavenging, reducing power and chelating iron ions. Also were tested against Bacillus cereus, Staphylococcus aureus, Escherichia coli, Psedomonas aeruginosa, Candida albicans, and Cryptococcus neoformans. In addition, cytotoxic activity was assayed against lung cancer A549 cell line. The methanol fraction showed no bioactivity but achieved the highest amount of phenolic, flavonol and flavonoid contents, also it showed a significant result as antioxidant, reducing power and chelating agent. The n-hexane fraction achieved the minimum inhibitory concentration (MIC) value 12.5 μg. mL(-1) against B. cereus while the MIC value for DCM fraction was 25 μg. mL(-1). The DCM fraction was more active against S. aureus where the result was 50 μg. mL(-1) while the n-hexane fraction was 100 μg. mL(-1). The three main fractions have shown no activity against gram negative bacterial and fungal. The n-hexane and DCM fractions have shown cytotoxicity against lung cancer cell line; the 50% inhibition concentration (IC(50)) was 22 ± 2.64 and 32 ± 3.78 μg. mL(-1) respectively. The results were statistically significant (P < 0.05). Among the results, C. canum fractions proved to be effective against gram positive bacterial and anti-proliferation activity. Also it showed antioxidant activity as well. The results provided beneficial information for communities as well as can help to search for alternative drugs, and will contribute to establish safe and effective use of phytomedicines in the treatment of diseases.
Fulltext Apoptosis, antimicrobial and antioxidant activities of phytochemicals from Garcinia malaccensis Hk.f

Taher M, Susanti D, Rezali MF, Zohri FS, Ichwan SJ, Alkhamaiseh SI, et al.

Asian Pac J Trop Med, 2012 Feb;5(2):136-41.
PMID: 22221758 DOI: 10.1016/S1995-7645(12)60012-1

OBJECTIVE: To study the chemical constituents of stembark of Garcinia malaccensis (G. malaccensis) together with apoptotic, antimicrobial and antioxidant activities.
METHODS: Purification and structure elucidation were carried out by chromatographic and spectroscopic techniques, respectively. MTT and trypan blue exclusion methods were performed to study the cytotoxic activity. Antibacterial activity was conducted by disc diffusion and microdilution methods, whereas antioxidant activities were done by ferric thiocyanate method and DPPH radical scavenging.
RESULTS: The phytochemical study led to the isolation of α,β-mangostin and cycloart-24-en-3β-ol. α-Mangostin exhibited cytotoxic activity against HSC-3 cells with an IC(50) of 0.33 μM. β- and α-mangostin showed activity against K562 cells with IC(50) of 0.40 μM and 0.48 μM, respectively. α-Mangostin was active against Gram-positive bacteria, Staphylococcus aureus (S. aureus) and Bacillus anthracis (B. anthracis) with inhibition zone and MIC value of (19 mm; 0.025 mg/mL) and (20 mm; 0.013 mg/mL), respectively. In antioxidant assay, α-mangostin exhibited activity as an inhibitor of lipid peroxidation.
CONCLUSIONS: G. malaccensis presence α- and β-mangostin and cycloart-24-en-3β-ol. β-Mangostin was found very active against HSC-3 cells and K562. The results suggest that mangostins derivatives have the potential to inhibit the growth of cancer cells by inducing apoptosis. In addition, α-and β-mangostin was found inhibit the growth of Gram-positive pathogenic bacteria and also showed the activity as an inhibitor of lipid peroxidation.
Critical role of ARID3B in the expression of pro-apoptotic p53-target genes and apoptosis

Pratama E, Tian X, Lestari W, Iseki S, Ichwan SJ, Ikeda MA

Biochem Biophys Res Commun, 2015 Dec;468(1-2):248-54.
PMID: 26519881 DOI: 10.1016/j.bbrc.2015.10.121

ARID3A and ARID3B are transcriptional targets of p53. Recently, it has been reported that ARID3A plays a critical role in the transcriptional activation of pro-arrest p21 in response to DNA damage. However, the role of ARID3B in the p53 regulatory pathway remains poorly understood. Here we show that ARID3A and ARID3B specifically bind to putative ARID3-binding sites in p53 target genes in vitro and in vivo. ARID3B and, to a lesser extent, ARID3A silencing blocked transcriptional activation of pro-apoptotic p53 target genes, such as PUMA, PIG3, and p53. Furthermore, ectopic ARID3B, to a lesser extent, ARID3A expression activated the pro-apoptotic gene expression, and only ARID3B induced apoptosis. Finally, ARID3B but not ARID3A silencing blocked apoptosis induction following DNA damage. These results indicated that, although ARID3B and ARID3A share overlapping functions, ARID3B play a key role in the expression of pro-apoptotic p53-target genes and apoptosis.
Intracellular biosynthesis of Au and Ag nanoparticles using ethanolic extract of Brassica oleracea L. and studies on their physicochemical and biological properties

Kuppusamy P, Ichwan SJ, Parine NR, Yusoff MM, Maniam GP, Govindan N

J Environ Sci (China), 2015 Mar 1;29:151-7.
PMID: 25766024 DOI: 10.1016/j.jes.2014.06.050

In this present study, we reported broccoli (Brassica oleracea L.) as a potential candidate for the synthesis of gold and silver nanoparticles (NPs) in green chemistry method. The synthesized metal nanoparticles are evaluated their antimicrobial efficacy against different human pathogenic organisms. The physico-chemical properties of gold nanoparticles were analyzed using different analytical techniques such as a UV-Vis spectrophotometer, Field Emission Scanning Electron Microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction and a Fourier Transform Infrared spectrophotometer. In addition, gold and silver NP antimicrobial efficacy was checked by disc diffusion assay. UV-Vis color intensity of the nanoparticles was shown at 540 and 450 nm for gold and silver nanoparticles respectively. Higher magnification of the Field Emission Scanning Electron Microscopy image shows the variable morphology of the gold nanoparticles such as spherical, rod and triangular shapes and silver nanoparticles were seen in spherical shapes. The average spherical size of the particles was observed in 24-38 nm for gold and 30-45 nm for silver NPs. X-ray diffraction pattern confirmed the presence of gold nanoparticles and silver nanoparticles which were crystalline in nature. Additionally, the functional metabolites were identified by the Fourier Transform Infrared spectroscopy. IR spectra revealed phenols, alcohols, aldehydes (sugar moieties), vitamins and proteins are present in the broccoli extract which are accountable to synthesize the nanoparticles. The synthesized gold and silver NPs inhibited the growth of the tested bacterial and fungal pathogens at the concentration of 50 μg/mL respectively. In addition, broccoli mediated gold and silver nanoparticles have shown potent antimicrobial activity against human pathogens.
A simple method for extracting both active oily and water soluble extract (WSE) from Nigella sativa (L.) seeds using a single solvent system

Babar ZM, Azizi WM, Ichwan SJ, Ahmed QU, Azad AK, Mawa I

Nat Prod Res, 2019 Aug;33(15):2266-2270.
PMID: 30037274 DOI: 10.1080/14786419.2018.1493587

The current study provides a way of extraction for both active NSO and WSE from Nigella sativa seeds using 98% methanol. About 1 kg of ground seeds was macerated by 1:2.5 w/v (g/mL) for 72 hours. After rotary evaporation and 7 days of continuous drying and chilling at 50 and 4 °C, NSO and WSE were obtained at the same instant. Solubility tests of 24 solvents and 11 thin layer chromatographic analyses while 2, 2-diphenyl-1-picrylhydrazyl free radical scavenging assay of NSO (73.66) , WSE (33.32) and NSO + WSE (78.22) against ascorbic acid (IC50 = 4.28 mg/mL) was performed. WSE was found to be highly soluble in water and 5% NaOH exhibiting the same Rf value of 0.95 for EtOH:DMSO (9:1) against the honey. WSE has revealed more than twofold higher anti-oxidant activity than others. Formulation of WSE with Tualang honey may provide better targeted hydrophilic drug delivery systems.
Fulltext Nutraceuticals as potential therapeutic agents for colon cancer: a review

Kuppusamy P, Yusoff MM, Maniam GP, Ichwan SJ, Soundharrajan I, Govindan N

Acta Pharm Sin B, 2014 Jun;4(3):173-81.
PMID: 26579381 DOI: 10.1016/j.apsb.2014.04.002

Colon cancer is a world-wide health problem and the second-most dangerous type of cancer, affecting both men and women. The modern diet and lifestyles, with high meat consumption and excessive alcohol use, along with limited physical activity has led to an increasing mortality rate for colon cancer worldwide. As a result, there is a need to develop novel and environmentally benign drug therapies for colon cancer. Currently, nutraceuticals play an increasingly important role in the treatment of various chronic diseases such as colon cancer, diabetes and Alzheimer׳s disease. Nutraceuticals are derived from various natural sources such as medicinal plants, marine organisms, vegetables and fruits. Nutraceuticals have shown the potential to reduce the risk of colon cancer and slow its progression. These dietary substances target different molecular aspects of colon cancer development. Accordingly, this review briefly discusses the medicinal importance of nutraceuticals and their ability to reduce the risk of colorectal carcinogenesis.
In Vitro Anticancer Activity of Au, Ag Nanoparticles Synthesized Using Commelina nudiflora L. Aqueous Extract Against HCT-116 Colon Cancer Cells

Kuppusamy P, Ichwan SJ, Al-Zikri PN, Suriyah WH, Soundharrajan I, Govindan N, et al.

Biol Trace Elem Res, 2016 Oct;173(2):297-305.
PMID: 26961292 DOI: 10.1007/s12011-016-0666-7

Recently, metal nanoparticles have been getting great medical and social interests due to their potential physico-chemical properties such as higher affinity, low molecular weight, and larger surface area. The biosynthesized gold and silver nanoparticles are spherical, triangular in shape with an average size of 24-150 nm as reported in our earlier studies. The biological properties of synthesized gold and silver nanoparticles are demonstrated in this paper. The different in vitro assays such as MTT, flow cytometry, and reverse transcription polymerase chain reaction (RT-qPCR) techniques were used to evaluate the in vitro anticancer properties of synthesized metal nanoparticles. The biosynthesized gold and silver nanoparticles have shown reduced cell viability and increased cytotoxicity in HCT-116 colon cancer cells with IC50 concentration of 200 and 100 μg/ml, respectively. The flow cytometry experiments revealed that the IC50 concentrations of gold and silver nanoparticle-treated cells that have significant changes were observed in the sub-G1 cell cycle phase compared with the positive control. Additionally, the relative messenger RNA (mRNA) gene expressions of HCT-116 cells were studied by RT-qPCR techniques. The pro-apoptotic genes such as PUMA (++), Caspase-3 (+), Caspase-8 (++), and Caspase-9 (++) were upregulated in the treated HCT-116 cells compared with cisplatin. Overall, these findings have proved that the synthesized gold and silver nanoparticles could be potent anti-colon cancer drugs.
Fulltext α-Mangostin Improves Glucose Uptake and Inhibits Adipocytes Differentiation in 3T3-L1 Cells via PPARγ, GLUT4, and Leptin Expressions

Taher M, Mohamed Amiroudine MZ, Tengku Zakaria TM, Susanti D, Ichwan SJ, Kaderi MA, et al.

Evid Based Complement Alternat Med, 2015;2015:740238.
PMID: 25873982 DOI: 10.1155/2015/740238

Obesity has been often associated with the occurrence of cardiovascular diseases, type 2 diabetes, and cancer. The development of obesity is also accompanied by significant differentiation of preadipocytes into adipocytes. In this study, we investigated the activity of α-mangostin, a major xanthone component isolated from the stem bark of G. malaccensis, on glucose uptake and adipocyte differentiation of 3T3-L1 cells focusing on PPARγ, GLUT4, and leptin expressions. α-Mangostin was found to inhibit cytoplasmic lipid accumulation and adipogenic differentiation. Cells treated with 50 μM of α-mangostin reduced intracellular fat accumulation dose-dependently up to 44.4% relative to MDI-treated cells. Analyses of 2-deoxy-D-[(3)H] glucose uptake activity showed that α-mangostin significantly improved the glucose uptake (P < 0.05) with highest activity found at 25 μM. In addition, α-mangostin increased the amount of free fatty acids (FFA) released. The highest glycerol release level was observed at 50 μM of α-mangostin. qRT-PCR analysis showed reduced lipid accumulation via inhibition of PPARγ gene expression. Induction of glucose uptake and free fatty acid release by α-mangostin were accompanied by increasing mRNA expression of GLUT4 and leptin. These evidences propose that α-mangostin might be possible candidate for the effective management of obesity in future.
In vivo anxiolytic and in vitro anti-inflammatory activities of water-soluble extract (WSE) of Nigella sativa (L.) seeds

Babar ZM, Jaswir I, Tareq AM, Ali Reza ASM, Azizi WM, Hafidz M, et al.

Nat Prod Res, 2021 Aug;35(16):2793-2798.
PMID: 31578877 DOI: 10.1080/14786419.2019.1667348

The WSE is a highly polar, gummy and mucilaginous bioactive content of the Nigella sativa (L.) seeds. This study reports the anxiolytic and anti-inflammatory effects of WSE investigated using Elevated Plus Maze (EPM) and Hole-Board Test (HBT) in adult mice and human RBCs haemolysis inhibition and protein denaturation respectively. The oral WSE treatment (100 & 200 mg/kg b.w/day) for 72 hours has exhibited slightly better anxiolytic effect (p