Protective Effects of Aqueous and Ethanolic Extracts of Portulaca oleracea L. Aerial Parts on H_2O_2- Induced DNA Damage in Lymphocytes by Comet Assay

Javad Behravan,Fatemeh Mosafa,Negar Soudmand,Elahe Taghiabadi,Bibi Marjan Razavi,Gholamreza Karimi 사단법인약침학회 2011 Journal of Acupuncture & Meridian Studies Vol.4 No.3

The comet assay is a standard method for measuring DNA damage. In this study, the protective effects of ethanolic and aqueous extracts of Portulaca oleracea L. (P. oleracea)on human lymphocyte DNA lesions were evaluated with the comet assay. Lymphocytes were isolated from blood samples taken from healthy volunteers. Human lymphocytes were incubated in H_2O_2 (50,100, and 200 mM), aqueous extract (0.05, 0.1,0.5, 1, and 2.5 mg/ml), and ethanolic extracts (0.05, 0.1, 0.5, 1, and 2.5 mg/ml) of P. oleraceae aerial parts alone with a combination of H_2O_2 (100 mM) with either 1 or 2.5 mg/ml of both extracts at 4℃ for 30 minutes. The extent of DNA migration was measured using the alkaline single cell gel electrophoresis approach assay, and DNA damage was expressed as percentage tail DNA. We found that the aqueous extract of P. oleracea significantly inhibited DNA damage, while there was no effect of the ethanolic extract. These data suggest that the aqueous extract of P. oleracea can prevent oxidative DNA damage to human lymphocytes, which is likely due to antioxidant constituents in the extract.

Cloning and Characterization of Directly Amplified Antiviral Gene Interferon Alpha-2b (HulFN$\{alpha}$-2b) from Human Leukocytes Chromosomal DNA

Behravan, Javad,Ahmadpour, Hassan The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.7

Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFN$\{alpha}$-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the HindIII and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.

Cloning and Characterization of Directly Amplified Antiviral Gene Interferon Alpha-2b (HuIFNα-2b) from Human Leukocytes Chromosomal DNA

Javad Behravan,Hassan Ahmadpour 대한약학회 2004 Archives of Pharmacal Research Vol.27 No.7

Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HuIFNa-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the HindIII and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.

Antigenotoxic Effects of Satureja hortensis L. on Rat Lymphocytes Exposed to Oxidative Stress

Fatemeh Mosaffa,Javad Behravan,Gholamreza Karimi,Mehrdad Iranshahi 대한약학회 2006 Archives of Pharmacal Research Vol.29 No.2

The protective properties of Satureja hortensis L. on the rat lymphocytes DNA lesions were tested. Lymphocytes were isolated from blood samples taken from healthy rats. DNA breaks and resistance to H2O2-induced damage were measured with the comet assay. Rat lymphocytes were incubated in S. hortensis ethanolic extract (SHE) (0.05, 0.1, 0.5, 1.0, and 2.5 mg/ mL), essential oil (SHEO)(0.05, 0.1, 0.5, 1.0, and 2.5 µL/mL), H2O2 (50, 100, and 200 µM), a combination of H2O2 (200 mM) with either SHE (1.0, 2.5 mg/mL) or SHEO (1.0, 2.5 µL/mL) at 4oC for 30 min, and the extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Treatment of rat lymphocytes with SHE or SHEO resulted in significant reduction of H2O2-induced DNA damage compared to controls. SHE exhibited a significant (P<0.01) inhibitory effect on oxidative DNA damage at 2.5 mg/mL. SHEO (1.0 and 2.5 µL/mL) also showed significant inhibitory effects (P <0.01) on H2O2 induced chromosomal damage. In conclusion both the ethanolic extract and the essential oil of the plant reversed the oxidative damage to rat lymphocytes induced by hydrogen peroxide.

Antigenotoxic Effects of Satureja hortensis L. on Rat Lymphocytes Exposed to Oxidative Stress

Mosaffa Fatemeh,Behravan Javad,Karimi Gholamreza,Iranshahi Mehrdad The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.2

The protective properties of Satureja hortensis L. on the rat lymphocytes DNA lesions were tested. Lymphocytes were isolated from blood samples taken from healthy rats. DNA breaks and resistance to $H_{2}O_{2}$-induced damage were measured with the comet assay. Rat lymphocytes were incubated in S. hortensis ethanolic extract (SHE) (0.05, 0.1, 0.5, 1.0, and 2.5 mg/mL), essential oil (SHEO)(0.05, 0.1, 0.5, 1.0, and 2.5 ${mu}L/mL$), $H_{2}O_{2}$ (50, 100, and 200 ${\mu}M$), a combination of $H_{2}O_{2}$ (200 mM) with either SHE (1.0, 2.5 mg/mL) or SHEO (1.0, 2.5 ${\mu}L/mL$) at $4^{\circ}C$ for 30 min, and the extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Treatment of rat lymphocytes with SHE or SHEO resulted in significant reduction of $H_{2}O_{2}$-induced DNA damage compared to controls. SHE exhibited a significant (P<0.01) inhibitory effect on oxidative DNA damage at 2.5 mg/mL. SHEO (1.0 and 2.5 ${\mu}L/mL$) also showed significant inhibitory effects (P<0.01) on $H_{2}O_{2}$ induced chromosomal damage. In conclusion both the ethanolic extract and the essential oil of the plant reversed the oxidative damage to rat lymphocytes induced by hydrogen peroxide.

Phorbol Ester TPA Modulates Chemoresistance in the Drug Sensitive Breast Cancer Cell Line MCF-7 by Inducing Expression of Drug Efflux Transporter ABCG2

Kalalinia, Fatemeh,Elahian, Fatemeh,Hassani, Mitra,Kasaeeian, Jamal,Behravan, Javad Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6

Recent studies have indicated a link between levels of cyclooxygenase-2 (COX-2) and development of the multidrug resistance (MDR) phenotype. The ATP-binding cassette sub-family G member 2 (ABCG2) is a major MDR-related transporter protein that is frequently overexpressed in cancer patients. In this study, we aimed to evaluate any positive correlation between COX-2 and ABCG2 gene expression using the COX-2 inducer 12-O-tetradecanoylphorbol-13-acetate (TPA) in human breast cancer cell lines. ABCG2 mRNA and protein expression was studied using real-time RT-PCR and flow cytometry, respectively. A significant increase of COX-2 mRNA expression (up to 11-fold by 4 h) was induced by TPA in MDA-MB-231 cells, this induction effect being lower in MCF-7 cells. TPA caused a considerable increase up to 9-fold in ABCG2 mRNA expression in parental MCF-7 cells, while it caused a small enhancement in ABCG2 expression up to 67 % by 4 h followed by a time-dependent decrease in ABCG2 mRNA expression in MDA-MB-231 cells. TPA treatment resulted in a slight increase of ABCG2 protein expression in MCF-7 cells, while a time-dependent decrease in ABCG2 protein expression was occurred in MDA-MB-231 cells. In conclusion, based on the observed effects of TPA in MDA-Mb-231 cells, it is proposed that TPA up-regulates ABCG2 expression in the drug sensitive MCF-7 breast cancer cell line through COX-2 unrelated pathways.

Cyclooxygenase-2 inhibition by novel Bisaryl imidazolyl imidazole derivatives increases Bax/Bcl-2 ratio and upregulates Caspase-3 gene expression in Caco-2 colorectal cancer cell line

Reza Entezari Heravi,Farzin Hadizadeh,Mojtaba Sankian,Jalil Tavakol Afshari,Javad Behravan 한국유전학회 2012 Genes & Genomics Vol.34 No.2

Cyclooxygenase-2 (COX-2) inhibitors including celecoxib inhibit cell growth and induce apoptosis in cancer cells. In this study, the relation of Bax (an apoptosis promoter) to Bcl-2(an apoptosis inhibitor) ratio with the apoptosis co-ordination enzyme, caspase-3 was investigated in correlation with the treatment of 4,5-bisaryl imidazolyl imidazoles as novel selective COX-2 inhibitors in Caco-2 colorectal cancer cells . Recently, the organic reactions under microwave irradiation attracted attention of scientists due to their high reaction rate,mild reaction conditions and the formation of clean products. Therefore, a microwave-assisted method was used to synthesize our compounds. The effects of these COX-2 inhibitors on the proliferation of Caco-2 cells were evaluated by MTT assay. cDNA microarray and clustering analysis were used to evaluate effects of our synthetic compounds on gene expression pattern of 112 genes involved in apoptosis pathways. Bax, Bcl-2 and caspase-3 mRNA expression and their relationship were analyzed by quantitative real-time PCR. Results indicated that proliferation of Caco-2 cells after treatment with 4,5-bisaryl imidazolyl imidazoles on Caco-2 cells were time and dose dependent. We conclude that increase in Bax/Bcl-2ratio leads to an up-regulation in caspase-3 mRNA expression.

Chemokine Receptors Expression in MSCs: Comparative Analysis in Diff erent Sources and Passages

Asieh Heirani-Tabasi,Shirin Toosi,Mahdi Mirahmadi,Mohammad Amir Mishan,Hamid Reza Bidkhori,Ahmad Reza Bahrami,Javad Behravan,Hojjat Naderi-Meshkin 한국조직공학과 재생의학회 2017 조직공학과 재생의학 Vol.14 No.5

MSC-based therapy is providing a cure for degenerative diseases with unmet medical need and usually iliac crest bone marrow (ICBM) are being applied in clinics. Alternative sources, including adipose tissue and reamer/irrigator/ aspirator hold great potential for isolating MCSs. Here, we compared original MSCs features of adipose tissue (Ad-MSCs) and bone marrow of long-bone (RIA-MSCs) or iliac crest, and the expression of chemokine receptors (including CXCR4, CX3CR1, CXCR6, CXCR2, CCR1 and CCR7) in these three sources, which are important in the context of homing. We further investigated the role of SDF-1/CXCR4 axis as a key player in motility of different population of MSCs using Transwell migration assay. All cells exhibited typical MSCs characteristics. However, different MSCs sources expressed different levels of chemokine receptors. Generally, the expression of these chemokine receptors was decreased with increasing passage (P) number from 2 to 3. Interestingly, it was observed that the CXCR4 expression and migration capacity in Ad-MSCs is significantly higher than ICBM and RIA-MSCs in P2. Although our data showed that CXCR4 had highest expression in P2 Ad-MSCs, but it dramatically declined following sub-culturing in the P3. Hence, to improve homing of MSCs by means of chemokine/their receptors axis, the source of isolation and passage number should be considered for clinical applications.