Investigation of the anticancer and antiangiogenic effects of hydroalcoholic extract of Shirazi thyme (Zataria multiflora) on rat C۶ glioma cell line and SPF embryonated egg chorioallantoic membrane (CAM) model

Glioma, as the most common primary malignant tumor of the central nervous system, is a major therapeutic challenge due to its high aggressiveness and resistance to conventional treatments. Tumor angiogenesis plays a pivotal role in the progression, proliferation, and metastasis of this tumor. In the last decade, traditional medicinal plants have been considered as rich sources of bioactive compounds with anticancer and antiangiogenic potential. Zataria multiflora, an endemic plant of the Iranian plateau with known antioxidant, anti-inflammatory, and antimicrobial activities, is a suitable candidate for evaluating antitumor effects.The aim of the present study was to investigate the cytotoxic (anticancer) and antiangiogenic effects of the hydroalcoholic extract of Zataria multiflora in in vitro and in vivo models. In the in vitro section, the rat glioma C۶ cell line was selected as a model. Cells were cultured at an initial density of ۵×۱۰³ to ۷×۱۰³ cells/well in a ۹۶-well plate and after ۲۴ hours of incubation, they were treated with concentrations of ۲۰, ۵۰, ۱۰۰, and ۲۰۰ μg/mL of extract. The negative control group contained only phosphate buffered saline. Cell viability was assessed ۴۸ hours after treatment by MTT assay. The results showed that the extract exerted cytotoxic activity in a dose-dependent manner; such that at concentrations of ۱۰۰ and ۲۰۰ μg/mL, a significant decrease in cell viability was observed compared to the control. In vivo, antiangiogenic effects were investigated using the SPF embryonated egg CAM model, which is a standard, ethical, and cost-effective model for studying angiogenesis. Three groups of ten were formed: control (PBS), ۱۰۰ μg treatment, and ۲۰۰ μg treatment. On the seventh day of embryonic development, ۱۰۰ μL of the respective solution was applied directly to the CAM membrane. Eggs were maintained under standard incubation conditions (۳۷°C, ۷۵% humidity) until day ۲۱, and vessels were monitored daily by candlelight. The results indicated a significant inhibition of angiogenesis at a dose of ۲۰۰ μg/mL, which was accompanied by a significant reduction in the number, diameter, and branching of vessels. At a dose of ۱۰۰ μg, milder inhibitory effects were observed.The overall findings of the study indicate a dose-dependent antiproliferative and antiangiogenic activity of Shirazi thyme extract. These effects are likely due to the presence of phenolic compounds, flavonoids, and monoterpenes present in the plant that are capable of interfering with cell growth signaling pathways, inducing apoptosis, and disrupting angiogenic factors. The present results highlight the therapeutic potential of Shirazi thyme as an adjuvant in the management of glioma and justify the need for future studies to isolate active compounds, elucidate molecular mechanisms, and evaluate in higher animal models.

کلیدواژه ها:

Shirazi thyme ، glioma ، antiangiogenesis ، MTT