Abbas Jafarian

Cytotoxicity of different extracts of arial parts of Ziziphus spina-christi on Hela and MDA-MB-468 tumor cells

Background: It has been shown that plants from the family Rhamnaceae possess anticancer activity. In this study, we sought to determine if Ziziphus spina-christi, a species from this family, has cytotoxic effect on cancer cell lines. Materials and Methods: Using maceration method, different extracts of leaves of Z. spina-christi were prepared. Hexane, chloroform, chloroform-methanol (9:1), methanol-water (7:1) methanol, butanol and water were used for extraction, after preliminary phytochemical analyses were done. The cytotoxic activity of the extracts against Hela and MDA-MB-468 tumor cells was evaluated by MTT assay. Briefly, cells were seeded in microplates and different concentrations of extracts were added. After incubation of cells for 72 h, their viability was evaluated by addition of tetrazolium salt solution. After 3 h medium was aspirated, dimethyl sulfoxide was added and absorbance was determined at 540 nm with an ELISA plate reader. Extracts were considered cytotoxic when more than 50% reduction on cell survival was observed. Results: Hexane, chloroform, chloroform-methanol, butanol, methanol-water and aqueous extracts of Z. spina-christi significantly and concentration-dependently reduced viability of Hela and MAD-MB-468 cells. In the both cell lines, chloroform-methanol extract of Z. spina-christi was more potent than the other extracts. Results: From the finding of this study it can be concluded that Z. spina-christi is a good candidate for further study for new cytotoxic agents.

Cytotoxic Evaluations of Iranian Conifers on Cancer Cells

ABSTRACT In this study, branchlets, fruit, or bark of Taxus baccata. L. as well as branchlets or fruits of two other species of Iranian conifers, namely, Platycladus orientalis. France and Cupressus sempervirens. L. var. horizentalis. (Mill) Gunde were collected, identified, and the cytotoxic effects of hydroalcoholic extracts on three human tumor cell lines were determined. Different concentrations of extracts were added to cultured cells and incubated for 72 h. Cell survival was evaluated using the MTT assay. Extracts from bark of female Taxus baccata. showed inhibitory activities against Hela cells. The extracts of the branchlets of male and female T. baccata. as well as obtained extract from fruits of P. orientalis. showed inhibitory activities against MDA-MB-468 cells, whereas the extracts of branchlets of female T. baccata. showed inhibitory activities against KB cells. In conclusion, obtained extract from bark of T. baccata. showed comparable cytotoxic effect to doxorubicin against Hela cells.

Comparing different sterol containing solid lipid nanoparticles for targeted delivery of quercetin in hepatocellular carcinoma

Abstract Quercetin (QT) is a potential chemotherapeutic drug with low solubility that seriously limits its clinical use. The aim of this study was enhancing cellular penetration of QT by sterol containing solid lipid nanoparticles (SLNs) which make bilayers fluent for targeting hepatocellular carcinoma cells. Three variables including sterol type (cholesterol, stigmasterol and stigmastanol), drug and sterol content were studied in a surface response D-optimal design for preparation of QT-SLNs by emulsification solvent evaporation method. The studied responses included particle size, zeta potential, drug loading capacity and 24 h release efficiency (RE24%). Scanning electron and atomic force microscopy were used to study the morphology of QT-SLNs and their thermal behavior was studied by DSC analysis. Cytotoxicity of QT-SLNs was determined by MTT assay on HepG-2 cells and cellular uptake by fluorescence microscopy method. Optimized QT-SLNs obtained from cholesterol and QT with the ratio of 2:1 that showed particle size of 78.0 ± 7.0 nm, zeta potential of −22.7 ± 1.3 mV, drug loading efficiency of 99.9 ± 0.5% and RE24 of 56.3 ± 3.4%. IC50 of QT in cholesterol SLNs was about six and two times less than free QT and phytosterol SLNs, respectively, and caused more accumulation of QT in HepG2 cells. Blank phytosterol SLNs were toxic on cells.

Immunization with a novel chimeric peptide representing B and T cell epitopes from HER2 extracellular domain (HER2 ECD) for breast cancer

Because of direct stimulating immune system against disease, vaccination or active immunotherapy is preferable compared to passive immunotherapy. For this purpose, a newly designed chimeric peptide containing epitopes for both B and T cells from HER2 ECD subdomain III was proposed. To evaluate the effects of the active immunization, a discontinuous B cell epitope peptide was selected based on average antigenicity by bioinformatics analysis. The selected peptide was collinearly synthesized as a chimera with a T helper epitope from the protein sequence of measles virus fusion (208-302) using the GPSL linker. Three mice were immunized with the chimeric peptide. Reactive antibodies with HER2 protein in ELISA and immunofluorescence assays with no cross-reactivity were generated. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay indicated that the anti-peptide sera had inhibitory effects on proliferation of SK-BR-3 cells. Hence, the newly designed, discontinuous chimeric peptide representing B and T cell epitopes from subdomain III of HER2-ECD can form the basis for future vaccines design, where these data can be applied for monoclonal antibody production targeting the distinct epitope of HER2 receptor compared to the two broadly used anti-HER2 monoclonal antibodies, Herceptin and pertuzumab.

Pneumonia associated with a single intratracheal dose of bleomycin in mice

Bleomycin-induced pulmonary fibrosis in mice is a commonly used animal model for human lung fibrosis1. For a study of bleomycin-induced pulmonary fibrosis, we obtained 30 female NMRI mice (25–30 g) from the Pasteur Institute (Tehran, Iran). Animals were housed in standard rodent cages in a mouse room on a 12-hr light/ dark cycle. The mice had ad libitum access to water and laboratory rodent chow. Experiments were carried out according to the International Guiding Principles for Biomedical Research Involv ing Animals as developed by the Council for International Organizations of Medical Sciences (CIOMS). To induce pulmonary fibrosis, we administered a single intratracheal dose of bleomycin to each mouse according to the protocol developed by Schrier et al.2. Under ketamine anesthesia (75 mg/kg i.p.), doses of 0.075 U bleomycin in 50 μl of sterile isotonic saline were instilled in mice intratracheally. Control mice were given the same volume of sterile saline. Six animals were used in each control and experimental group. Each week for four weeks after bleomycin instillation, we euthanized a group of 6 mice by ketamine overdose, removed the lungs, and perfused the left lung lobe with 10% neutral buffered formalin (NBF) solution via the trachea. The perfused lungs were then fixed in a 10% NBF solution before processing for histological examination. After sectioning and histological processing, sections of lung were stained with hematoxylin and eosin (H & E) or Masson’s trichrome and examined by light microscopy. Some lung sections showed multifocal foamy macrophages at three and four weeks after treatment with the single intratracheal dose of bleomycin. One case had histological changes involving the appearance of intra-alveolar and interstitial accumulation of lipid-laden, foamy macrophages (Fig. 1). There was also severe distortion of the pulmonary cytoarchitectural structure and multifocal fibrotic areas characterized by fibrous thickening

Evaluation of Cytotoxic Effect of Different Extracts of Seidlitzia rosmarinus on HeLa and HepG2 Cell Lines

Background: Seidlitzia rosmarinus which is commonly called “Oshnan” or “Eshnan” in Persian belongs to Chenopodiaceae family. Conventionally, it is believed that this plant is toxic. This study was aimed to evaluate the cytotoxic effect of S. rosmarinus against HeLa and HepG2 cell lines. Materials and Methods: S. rosmarinus was collected from the desert near Yazd, Iran. Hexane, chloroform, chloroform/methanol (9:1), and butanol extracts of aerial parts of S. rosmarinus were prepared. Doxorubicin and dimethyl sulfoxide 10% were used as positive and negative control, respectively. The cytotoxic activity was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: All extracts significantly and concentration dependently reduced viability of HeLa and HepG2 cells. Hexane, chloroform, and butanol extracts at doses of 200, 500, 750, and 1000 μg/ml significantly reduced HeLa cell viability (P < 0.05). Chloroform/methanol extract at doses of 100–500 μg/ml significantly reduced HeLa cell viability (P < 0.05). Hexane, chloroform, and butanol extracts at doses of 500, 750, and 1000 μg/ml significantly reduced HepG2 cell viability (P < 0.05). Chloroform/methanol extract at doses of 200, 300, 400, and 500 μg/ml significantly reduced HepG cell viability (P < 0.05). The most cytotoxic extract was chloroform/methanol extract in both cell lines. Furthermore, in the both cell lines, the second potent extract was chloroform extract. Conclusions: It can be concluded from the findings of this study that S. rosmarinus is a good candidate for further study to find new cytotoxic agents. Phytochemical investigation on chloroform/methanol extract and their structures is recommended.

Phytochemical Screening and Cytotoxic Evaluation of Euphorbia turcomanica on Hela and HT-29 Tumor Cell Lines

Background: Cancer is a term for a large group of different diseases, all involving uncontrolled cell growth. Many of Euphorbiaceae plants have been traditionally used for the treatment of ulcers, tumors, warts, and other diseases. In addition, in the last decade, there are studies showing cytotoxic effects of different species of Euphorbia on tumor cell lines. In this study, we attempted to determine if Euphorbia turcomanica possess any cytotoxic activity. Materials and Methods: Solvents extracted the plant powder with various polarities by a maceration method, and qualitative phytochemical analyzes were carried out on them to identify the constituents. On the other hand, the possible cytotoxicity of different extracts on Hela and HT-29 tumor cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and 50% reduction in cell survival was considered as a cytotoxic effect. Analyze of variance followed by Student-Newman-Keuls test was used to see the differences among the groups. Results: Phytochemical analysis of E. turcomanica showed the presence of flavonoid, alkaloid, anthraquinone and tannin in plant aerial parts. Methanol-water, acetone, dichloromethane, methanol, and heptane extracts of E. turcomanica significantly reduced viability of Hela cells (P < 0.05) with inhibitory concentration 50% (IC50) of 50, 90, 230, 420, and 450 μg/ml, respectively. While methanol-water, dichloromethane, methanol, ethyl acetate, and heptane extracts were cytotoxic with IC50of 43, 115, 125, 250, and 390 μg/ml, respectively (P < 0.05), on HT-29 cells. Conclusion: It can be concluded that E. turcomanica is a good candidate for further study toward cytotoxic agents.