ABSTRACT
Salvia chorassanica Bunge is one of the Iranian endemic species of Salvia. There is not any reported literature on S. chorassanica. This study was designed to examine the in-vitro anti- proliferative and proapoptotic effects of the methanol extract of S. chorassanica and its fractions on HeLa cell line. Cells were cultured in EX-CELL®, an animal free medium specially designed for HeLa cell line and incubated with different concentrations of plant extracts. Cell viability was quantified by MTS assay. Apoptotic cells were determined using propidium iodide [PI] staining of DNA fragmentation by flow cytometry [sub-G1 peak]. Activity of caspase -3, -8 and -9 was measured by the caspase colorimetric kit assay. S. chorassanica inhibited the growth of malignant cells and the CH[2]Cl[2] fraction was determined as the most cytotoxic fraction in comparison with other fractions. The calculated IC[50] values for methanol extract, n-hexane, and CH[2]Cl[2] and EtOAc fractions were 8.841, 5.45, 2.38, and 58.03 [micro]g/mL, respectively. S. chorassanica induced a sub-G1 peak in the flow cytometry histogram of treated cells compared to control cells indicating that the cytotoxic mechanism is characterized by apoptosis induction. The activity of caspase-3 and 8 proteins in treated HeLa cells was significantly higher than that of the control while caspase-9 activity did not change significantly. Based on the result obtained from our study, the apoptosis pathway involved in S. chorassanica-induced cell death may be through the extrinsic pathway and it can be a novel promising candidate in the treatment of cancer
ABSTRACT
The discovery and development of natural products with potent antioxidant properties has been one of the most interesting and promising approaches in the search for treatment of CNS injuries. The most significant consequence of the oxidative stress is thought to be the DNA modifications, which can become permanent via the formation of mutations and other types of genomic instability resulting cellular dysfunction. Serum/glucose deprivation [SGD] has served as an excellent in vitro model for the understanding of the molecular mechanisms of neuronal damage during ischemia and for the development of neuroprotective drugs against ischemia-induced brain injury. Nigella sativa [N. sativa] seeds and thymoquinone [TQ], its most abundant constituent, have been shown to possess antiinflammatory, antioxidant, chemopreventive and anti-neoplastic effects both in vitro and in vivo. Therefore, in this study we investigated genoprotective effects of N. sativa and TQ on DNA damage of PC12 cells under SGD condition. PC12 cells were cultured in DMEM medium containing 10% [v/v] fetal bovine serum, 100 units/ml penicillin, and 100 micro g/ml streptomycin. Initially cells were pretreated with different concentrations of N. sativa extract [NSE], [10, 50, 250 micro g/ml] and TQ [1, 5, 10 micro g/ml] for 6 h and then deprived of serum/glucose [SGD] for 18 h. The alkaline comet assay was used to evaluate the effect of these compounds on DNA damage following ischemic insult. The amount of DNA in the comet tail [% tail DNA] was measured as an indicator of DNA damage. A significant increase in the% tail DNA was seen in nuclei of cells following SGD induced DNA damage [p<0.001]. In the control groups, no significant difference was found in the% tail DNA between NSE- or TQ-pretreated and vehicle-pretreated PC12 cells [p>0.05]. NSE and TQ pretreatment resulted in a significant decrease in DNA damage following ischemic insult [p<0.001]. This suppression of DNA damage by NSE and TQ was found to be dose-dependent. These data indicate that NSE and TQ have a genoprotective property, as revealed by the comet assay, under SGD condition in PC12 cells
ABSTRACT
Teucrium polium has been known as an important traditional medicinal plant and is used for different therapeutic purposes such as gastrointestinal disorders. Therefore, the anti-spasmodic and anti-nociceptive activities of aqueous extract of Teucrium polium was examined. Anti-spasmodic effect of different concentrations [47-470 mg/1] of Teucrium polium extract was assessed on acetylcholine [220 nM] precontracted guinea pig isolated ileum. The anti-cholinergic effect of the plant was also examined by obtaining concentration-response curves in the absence and presence of Teucrium polium extract [470 mg/1] and atropine [10 nM]. Anti-nociceptive effect of different doses [30-240 mg/kg] of Teucrium polium aqueous extract was determined by hot-plate test on mice and compared with the effect of morphine [10 mg/kg] as positive control. Maximum inhibition response induced by Teucrium polium extract on contraction induced by acetylcholine [220 nM] was 93.5%. In the absence and presence of Teucrium polium extract [470 mg/I] and atropine [10 nM] the EC50 [the effective concentration causing 50% of maximum response] of Ach were 28.3 +/- 2.1, 55.4 +/- 3.7 and 208.1 +/- 9.2 nM respectively. There was also a parallel right-ward shift in the log concentration-response curve of acetylcholine in the presence of atropine, but a non-parallel shift in the presence of Teucrium polium extract. The Teucrium polium extract increased reaction time dose-dependently [P<0.01 for all doses]. However the anti-nociceptive effect of extract was significantly less than that of morphine [P<0.001]. These results show that Teucrium polium aqueous extract have anti-nociceptive and anti-spasmodic effects and may have some clinical benefits for gastrointestinal disorders