Cytotoxic Properties of Anthraquinones (Nordamnacanthal and Damnacanthal) from Roots of Morinda Elliptica

The study on the cytotoxic properties of nordamnacanthal and damnacanthal, the anthraquinones isolated from the roots of Morinda elliptica (family Rubiaceae) were carried out on several cancerous cell lines including CEM-SS (T-lymphoblastic leukaemia), KU812F (chronic myelogeneous leukaemia), WEHI-3 (leukaemia), HT29 (colon cancer) and HeLa (cervical adenocarcinoma). The degree of cytotoxicity of the compounds were defined by their abilities at certain concentration to cause 50% reduction in cell number relative to the untreated sample, and termed as IC50 value. CEM-SS was observed to be the most sensitive cell line towards nordamnacanthal and damnacanthal with the ICso values of 1.7 µg/ml and 10 µg/ml, respectively, as detected by the colorimetric tetrazolium-based assay (MTT). The compounds also showed cytotoxicity to the non-cancerous cell lines such as HF19 (lung fibroblast), human peripheral blood mononuclear (PBMC), 3T3 (mouse embryo) and Vero (monkey kidney fibroblast) but at very high concentrations (>30 µg/ml). Themicroscopic analysis on the treated CEM-SS cells including light m icroscopy without staining or following staining with haematology polychrome, Giemsa and Wright's stains, fluorescence microscopy following staining with acridine orange and propidium iodide, and scanning and transmission electron microscopy showed that these compounds induced two types of cell death, apoptosis and necrosis. At the molecular level, these compounds caused intemucleosomal DNA cleavage producing multiple of 180-200 bp fragments that are visible as a "ladder" on the agarose gel. The DNA fragmentation has been found to be due the activation of the Mg2+/Ca2+-dependent endonuclease. The induction of apoptosis by nordamnacanthal was different from the one induced by damnacanthal in a way that it occurs independently of ongoing transcription process. Nevertheless, in both cases, the process of dephosphorylation of protein phosphates 1 and 2A, the ongoing protein synthesis and the elevations of the cytosolic Ca2+ concentration were not needed for apoptosis to take place. Nordamnacanthal and damnacanthal at their ICso values showed different mechanism by which they exert their cytotoxic effects. Nordamnacanthal was found to have cytotoxic effect by inducing apoptosis in CEMSS cells. Damnacanthal, on the other hand, showed cytostatic effect by causing arrest at the GO/G1 phase of the cell cycle. Nordamnacanthal was also found to reduce the expression of bcl-2, thus stimulating the process of apoptosis in CEM-SS cells

Dillenia species: a review of the traditional uses, active constituents and pharmacological properties from pre-clinical studies

Context: Dillenia (Dilleniaceae) is a genus of about 100 species of flowering plants in tropical and subtropical trees of Southern Asia, Australasia, and the Indian Ocean Islands. Until now, only eight Dillenia species have been reported to be used traditionally in different countries for various medical purposes. Out of eight species, D. pentagyna (Roxb), D. indica (Linn.) and D. suffruticosa (Griffith Ex. Hook. F. & Thomsom Martelli) have been reported to be used to treat cancerous growth. Objective: The present review explored and provided information on the therapeutic potential of Dillenia species. Methods: Comprehensive and relevant literature on the therapeutic potential of Dillenia species was gathered through electronic databases including Google Scholar, Scopus, PubMed, and books, without limiting the dates of publication. Results and conclusion: The review demonstrated that only a few Dillenia species have been proven scientifically for their therapeutic potential in pre-clinical studies, including D. pentagyna, D. indica, D. papuana (Martelli), D. meliosmifolia (Hook. F. Ex. Thomsom) and D. suffruticosa (Griffith Ex Hook. F. & Thomson). A few species of Dillenia have undergone isolation and characterization of compounds with lupeol and betulinic acids having tremendous pharmacological potential. Dillenia species warrant further studies on their therapeutic potential, which may eventually lead to the development of new drug candidates for treatment of various diseases

Thymoquinone from Nigella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein

Thymoquinone (TQ), the active constituent of Nigella sativa or black cumin exhibited cytotoxic effects in several cancer cell lines. In this study, the cytotoxicity of TQ in human cervical squamous carcinoma cells (SiHa) was investigated. TQ was cytotoxic towards SiHa cells with IC50 values of 10.67 ± 0.12 and 9.33 ± 0.19 μg/mL as determined by MTT assay and trypan blue dye exclusion test, respectively, after 72 h of incubation. TQ was more cytotoxic towards SiHa cells compared to cisplatin. Interestingly, TQ was less cytotoxic towards the normal cells (3T3-L1 and Vero). Cell cycle analysis performed by flowcytometer showed a significant increase in the accumulation of TQ-treated cells at sub-G1 phase, indicating induction of apoptosis by the compound. Apoptosis induction by TQ was further confirmed by Annexin V/PI and AO/PI staining. Significant elevation of p53 and down-regulation of the anti-apoptotic Bcl-2 protein was found in the treated cells, without any changes in the expression of the pro-apoptotic Bax protein. In conclusion, thymoquinone from N. sativa was more potent than cisplatin in elimination of SiHa cells via apoptosis with down-regulation of Bcl-2 protein

Effects of thymoquinone rich fraction and thymoquinone on plasma lipoprotein levels and hepatic low density lipoprotein receptor and 3-hydroxy-3-methylglutaryl coenzyme A reductase genes expression.

The hypocholesterolemic effect of thymoquinone rich fraction (TQRF) extracted from Nigella sativa seeds using supercritical fluid extraction (SFE) in comparison with commercial available thymoquinone (TQ) in male Sprague–Dawley rats was investigated. Rats were fed prepared diet supplemented with 1% (w/w) cholesterol and treated with TQRF at, 0.5, 1 and 1.5 g/kg and TQ at 20, 50 and 100 mg/kg for 8 weeks. Plasma total cholesterol levels (TC) and low density lipoprotein cholesterol (LDLC) were significantly decreased in the TQRF and TQ treated rats compared to untreated rats. mRNA level of low density lipoprotein receptor (LDLR) was significantly expressed and the mRNA level of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-COAR) was significantly suppressed in the TQRF and TQ treated rats at different doses compared to untreated rats. These new findings identify TQRF and TQ as natural cholesterol lowering agents, and our study provides a molecular basis for the mechanisms of action through regulation of cholesterol in two main mechanisms first, uptake of LDLC via up regulation of LDLR gene and second, inhibition the synthesis of cholesterol via suppressing the HMG-COAR gene

The improvement of in vivo model (Balb/c mice) for cervical carcinogenesis using diethylstilbestrol (DES)

Cervical cancer is the most common gynecological cancer and one of the major causes of female cancer-related death worldwide particularly in developing countries. Thus far, there are a few in vivo models have been developed in investigating this type of cancer. In this study, we induced cervical cancer in Balb/c mice by exploiting the carcinogenic property of diestylstilbestrol (DES). The Balb/c pregnant mice were given subcutaneous (SC) injection of 67 μg/kg body weight of DES on GD 13, and the mice gave birth approximately at gestation day 19-22. Female offspring were reared and the body weight was recorded once weekly. The female offspring were sacrificed at age of 5 months. Upon termination, blood was collected in a plain tube via cardiac puncture and the reproductive tracts were collected and weighed. The reproductive tract sections were stained using H&E for observation of pathological changes. The progression of disease state was monitored by measuring the level of serum interleukin (IL-6) using the Mouse IL-6 ELISA Assay Kit (BD OptEIA™, USA). All parameters were compared with Not-induced group. The outcome of this study demonstrated a significant difference in body weight gain, reproductive organ weight, diameter of cervix and the level of serum IL-6 in the Induced group as compared to the Not-induced group (P< 0.05). Histopathological findings revealed the presence of adenosis only in the Induced group. It shows that DES could be employed as an agent to induce cervical carcinogenesis in animal model. In addition to that, new potential anti-cancer agents from various sources could be further evaluated using this technique

Toxicology study of vanillin on rats via oral and intra-peritoneal administration.

Vanillin is useful as anti-sickle cell anemia, anti-mutagen and anti-bacteria agent. However, vanillin must be administered at high concentration and cannot be oxidized by the upper gastrointestinal track of patients to be medically effective. In this study, we assessed the toxic effect of vanillin when administered in an un-oxidized form at high concentrations (150 and 300 mg/kg) via oral and intra-peritoneal injection. It was found that 300 mg/kg vanillin injection caused the rats to be unconscious without exerting any toxic effect on blood cells, kidney and liver. Besides, it showed blood protective property. Further analysis with GenomeLab GeXP genetic system on brain tissues showed that the expression of most xenobiotic metabolism, cell progression, tumor suppressor, DNA damage and inflammation genes were maintained at normal level. However, the expression of a few xenobiotic metabolism, cell cycle arrest and apoptosis genes were up-regulated by 5% ethanol injection. Nevertheless, when 5% ethanol was injected with the presence of vanillin, the expression was back to normal level. It is postulated that vanillin might have neuro-protective property. In conclusion, vanillin is not toxic at high concentration in both oral and intra-peritoneal injection and could provide blood and brain protective properties

Nordamnacanthal induced apoptosis and mitotic-G2/M arrest with downregulation of Bcl-2 in the human breast cancer cell line (MCF-7).

Nordamnacanthal, an anthraquinone extracted from the root of Morindaelliptica from Rubiaceae family has cytotoxic properties towards cancer cell lines and anti tumor promoting activities. This study was conducted to determine the cytotoxic effect of nordamnacanthal towards MOLT-4 and MCF-7 cell lines. Nordamnacanthal was found to be more cytotoxic towards MOLT-4 than MCF-7 with the IC50 of 3.8 μg/ml and 54μg/ml, respectively, as detected by using the trypan blue dye exclusion test. The nordamnacanthal-treated cells showed characteristics of apoptosis such as membrane blebbing, chromatin condensation and formation of apoptotic bodies as observed under an inverted lightmicroscope. Fluorescence analysis of cell death using acridine orange and propidium iodide staining showed that the population of MOLT-4 and MCF-7 cells underwent apoptosis at the IC50 value was 32% and 30.4%,respectively. Cell cycle analysis by flow cytometry indicated that nordamnacanthal did arrest MCF-7 cells at the G2/M phase. For MOLT-4, no cell cycle arrest was observed. Bcl-2 and Bax were downregulated in nordamnacanthal-treated MCF-7 cells. On the other hand, expression of the proteins in MOLT-4 was not from the control. In conclusion, nordamnacanthal was more cytotoxic towards MOLT-4 than MCF-7 cell line. The compound induced apoptosis in both cell lines, but with G2/M arrest and the involvement of Bcl-2 and Bax only in MCF-7

Aedestech Mosquito Home System prevents the hatch of Aedes mosquito eggs and reduces its population

Dengue fever (DF) is a global health problem and considered to be endemic in Malaysia. Conventional mosquito traps currently applied as vector control do not effectively reduce Aedes mosquito population. AedesTech Mosquito Home System (AMHS) is an autocidal ovitraps for Aedes mosquitoes that uses the ‘lure and kill’ concept and is expected to be able to reduce Aedes mosquito population. The effectiveness of AMHS in reducing Aedes mosquito population was investigated in Block A, B and D (control) of the 17th College, Universiti Putra Malaysia (UPM). For the first two weeks (pre-intervention), the conventional ovitraps were used to obtain the initial abundance of mosquito population in Block A, B and D. Subsequently, AMHS was used for the next three months and again followed by the conventional ovitrap for the final two weeks (post-intervention). Ovitrap Index, Hatching Index and percentage of emergence of adult mosquitoes were calculated once every two weeks. Data were analysed using Paired Sample T-test. Values were considered significant at p≤0.05. The Ovitrap Index that indicates the mosquito population at Block A and B was significantly higher (p≤0.05) than of Block D. Hatching Index of AMHS was significantly lower (p≤0.05) then conventional ovitraps. All mosquito eggs collected in AMHS did not develop into adult mosquitoes. There was a significant reduction (p≤0.05) in the mosquito population between the pre- and post-intervention. In conclusion, AMHS was effective in reducing the mosquito population in 17th College, UPM. Therefore, it is believed to be a very promising vector management option to control the incidence of DF

Bcl-2 was downregulated in G2/M-arrest breast cancer cells MCF-7-treated with nordamnacanthal

Nordamnacanthal, an anthraquinone extracted from the root of Morinda elliptica has cytotoxic properties towards various cancer cell lines and antitumor-promoting activities. This study was conducted to determine the effects of nordamnacanthal on the cell cycle, and the expression of Bcl-2 and Bax in breast cancer (MCF-7) and acute t-lymphoblastic leukemia (MOLT-4) cells at 50% of the total cell population underwent apoptosis. Nordamnacanthal caused 50% of MCF-7 and MOLT-4 cells underwent apoptosis at 15 μg/ml and 70 μg/ml, respectively, as analyzed by using a fluorescence microscope following staining with the acridine orange (AO) and propidium iodide (PI). The apoptotic cells exhibited nuclear fragmentation, chromatin condensation in the nucleus and membrane blebbing. Cell cycle analysis by flow cytometry indicated that nordamnacanthal arrested MCF-7 cells at the G2/M phase. For MOLT-4, no cell cycle arrest was observed. Bcl-2 and Bax were downregulated in nordamnacanthal-treated MCF-7 cells. On the other hand, expression of the proteins in MOLT-4 was not significantly different (p>0.05) from the control. In conclusion, treatment of MCF-7 cells with nordamnacanthal at the concentration that caused 50% of the total cell population underwent apoptosis, induced the G2/M arrest with downregulation of Bcl-2