Cytotoxic Effect of Saffron Stigma Aqueous Extract on Human Prostate Cancer and Mouse Fibroblast Cell Lines.

PURPOSE: Several lines of experimental evidence have shown that saffron has anticarcinogenic effects. This study aimed at evaluating the possible anticancer effect of saffron stigma aqueous extract on human prostate cancer (PC3) and mouse fibroblast cells (L929) as non-cancerous control cells. MATERIALS AND METHODS: Saffron stigma aqueous extract at concentrations of 100, 200, 400, 600, 800, 1600 and 3200 µg/mL were prepared. PC3 and L929 cells were incubated with different concentrations of saffron extracts in different time intervals (24, 48, 72, 96 and 144 hours). MTT assay was used for each cell line to investigate the cytotoxic effect of saffron. Morphological alterations were also observed under light inverted microscope. RESULTS: In fibroblast cell line after 24 hours, Saffron extract did not affect significantly the normal cells and they were intact in morphologic view. After 96 hours in the cells with highest concentration (1600 µg/mL), cell death and cellular form changes as well as severe granulation was observed. In prostate cell line after 24 hours, the only changes were observed in cells with the concentration of 1600 µg/mL. The cells were granulated and the form of the cells were spherule. After 72 hours, in group with the concentration of 1600 µg/mL, severe granulation was observed and the cell count decreased and some cells were dead. CONCLUSION: Saffron aqueous extract has an in vitro inhibitory effect on the proliferation of human prostate cell and mouse L929 cells which is dose-dependent.

Immunoregulatory Effects of Krocina™, a Herbal Medicine Made of Crocin, on Osteoarthritis Patients: A Successful Clinical Trial in Iran.

Osteoarthritis (OA) is the major cause of joint pain and disability. This research was planned to examine the effects of Krocina™, aherbal medicine made of crocin, an ingredient of saffron, in patients with OA. Forty patients suffering from OA were enrolled in our study and randomly divided into two groups, receiving Krocina™ and placebo, and the clinical trial continued for four months.Peripheral blood was taken from all patients and the percentage ofvarious subsets of T cells in addition to the levels of forkhead box protein P3 (FOXP3) and interleukin (IL)-17 were measured by flow cytometry technique. The visualan alog scale (VAS) index analysis decreased significantly in both groups (krocinaTM and placebo) (p<0.05). Assessment of the C-reactive protein (CRP) level in serum showed a significant decrease in the krocinaTM group (p<0.05). Moreover, we found a meaningful increase in the percentage of regulatory T cells (Tregs)cellin samples gathered from Krocina™ group (P=0.02) patients. The mean percentages of T helper (Th) 17 cellsinthe Krocina™ group and CD8+ T cellsin the placebo group patients were also meaningfully reduced (p<0.05). The geometric mean fluorescence intensity (GMFI) for IL-17 showed a significant decrease and increase in Krocina™ and placebo groups, respectively (p<0.05). No noticeable difference was observed in the percentages of Th cells and GMFI-FOXP3 in either group. Treg/Th17 ratio was shifted towards Tregscell in Krocina™ group at the end of the intervention. It is concluded that Krocina™ has immunoregulatory effects on patients with OA, ameliorating the disease.

Significant immunomodulatory properties of curcumin in patients with osteoarthritis; a successful clinical trial in Iran.

Osteoarthritis (OA) routinely is known as a multifactorial degenerative joint disease. This trial aimed to assess the curcumin (an active element of turmeric) effects on the immune responses in OA patients. Thirty patients were selected according to the American College of Rheumatology (ACR) criteria and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and equally divided into the two groups; intervention (received Sinacurcumin® 80 mg daily) and placebo, followed for 3 months. In the intervention group, our data showed a noticeably decrease in Visual Analog Score (VAS), C-reactive protein (CRP), CD4+ and CD8+ T cells, Th17 cells and B cells frequency. Additionally, Treg cells indicated a significant increase and Treg/Th17 cells ratio showed a meaningfully shifted toward Treg lymphocytes. In conclusion, our data indicated that clinical manifestation was ameliorated considerably following the administration of curcumin. Moreover, our data demonstrated the immunomodulatory effects of curcumin in OA patients.

Comparison of Th1/Th2 and Treg/Th17 ratios between wet and dry cupping therapies in Persian medicine.

OBJECTIVE: In Persian medicine (PM), wet-cupping therapy (WCT) is the most utilized approach. WCT is mostly done between the shoulders, which is referred to as "hejamt-e-aam" in the Persian language. CD4+T cells also refer to T helper lymphocytes play a critical role in the immune system. Naïve CD4+ T cells differentiate into at least four subsets, T helper 1 (Th1), T helper 2 (Th2), T helper 17 (Th17), and T regulatory (Treg) cells. The master regulator controlling each subset have been defined as follows, Tbet (Th1), Gata3 (Th2), RORγt (Th17), FoxP3 (Treg). The purpose of this study was to compare the effect of WCT and dry-cupping therapy (DCT) on the ratios of Th1/Th2 and Treg /Th17 in healthy individuals. MATERIALS AND METHODS: Participants were divided randomly into two groups of 41 men in the WCT group and 40 men in the DCT group. Blood was taken, before, one and four weeks after the intervention. RNA was extracted from the peripheral blood mononuclear cells and the expression of T-bet, GATA-3, RORγt, and Foxp3 genes were determined by using SYBR green RT-PCR technique. RESULTS: The results showed that WCT increased the expression ofGATA-3, RORγt, and Foxp3 transcription factor genes (p=0.009, p=0.001, and p=0.021, respectively). Although in the WCT group, the ratio of Foxp3/RORγt increased (p=0.048), but the ratio of T-bet/GATA-3 (Th1/Th2) decreased (p=0.971). CONCLUSION: Our findings indicated that WCT may regulate the T subsets of lymphocyte and reduce inflammation.

Suppression of pulmonary tumor promotion and induction of apoptosis by Crocus sativus L. extraction.

Crocus sativus L., commonly known as saffron, is the raw material for one of the most expensive spice in the world, and it has been used in folk medicine for centuries. We investigated the potential of the ethanolic extract of saffron to induce cytotoxic and apoptosis effects in carcinomic human alveolar basal epithelial cells (A549), a commonly used cell culture system for in vitro studies on lung cancer. The cells were cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum treated with different concentrations of the ethanolic extract of saffron for two consecutive days. Cell viability was quantitated by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Apoptotic cells were determined using annexin V-fluorescein isothiocyanate by flow cytometry. Saffron could decrease the cell viability in the malignant cells as a concentration- and time-dependent manner. The IC50 values against the A549 cell lines were determined as 1,200 and 650 µg/ml after 24 and 48 h, respectively. Saffron-induced apoptosis of the A549 cells in a concentration-dependent manner, as determined by flow cytometry histogram of treated cells that induced apoptotic cell death, is involved in the toxicity of saffron. It might be concluded that saffron could cause cell death in the A549 cells, in which apoptosis plays an important role. Saffron could also be considered as a promising chemotherapeutic agent in lung cancer treatment in future.

Role of caspases and Bax protein in saffron-induced apoptosis in MCF-7 cells.

Saffron (Crocus sativus), widely used as a spice in Middle Eastern cuisine and is known for anti-cancer properties. The mechanism of saffron-induced cytotoxicity, in tumor cells has not been adequately explored. Therefore, we investigated the role of caspases and Bax protein in saffron-induced apoptosis in MCF-7 cells, a commonly used cell culture system for in vitro studies on breast cancer. Cells were incubated with different concentrations of saffron extract. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). Role of caspase were studied using the pan-caspase inhibitor. Bax protein expression was analysed by western blotting. Saffron extract (200-2000 microg/ml) decreased cell viability in MCF-7 cells as a concentration- and time-dependent manner with an IC50 of 400+/-18.5 microg/ml after 48 h. Analysis of DNA fragmentation by flow cytometry showed apoptotic cell death in MCF-7 cell treated with saffron extract. Saffron-induced apoptosis could be inhibited by pan-caspase inhibitors, indicating caspase-dependent pathway was induced by saffron in MCF-7 cells. Bax protein expression was also increased in saffron-treated cells. Thus saffron exerts proapoptotic effects in a breast cancer-derived cell line and could be considered as a potential chemotherapeutic agent in breast cancer.

Study of cytotoxic and apoptogenic properties of saffron extract in human cancer cell lines.

Saffron (dried stigmas of Crocus sativus L.) has been used as a spice, food colorant and medicinal plant for millennia. In this study cytotoxic effect of saffron extract was evaluated in HepG2 and HeLa cell lines. Meanwhile role of apoptosis and ROS were explored. Malignant and non-malignant cells (L929) were cultured in DMEM medium and incubated with different concentrations of ethanolic saffron extract. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). ROS was measured using DCF-DA by flow cytometry analysis. Saffron could decrease cell viability in malignant cells as a concentration and time-dependent manner. The IC50 values against HeLa and HepG2 were determined 800 and 950 microg/ml after 48 h, respectively. Saffron induced a sub-G1 peak in flow cytometry histogram of treated cells compared to control indicating apoptotic cell death is involved in saffron toxicity. This toxicity was also independent of ROS production. It might be concluded that saffron could cause cell death in HeLa and HepG2 cells, in which apoptosis or programmed cell death plays an important role. Saffron could be also considered as a promising chemotherapeutic agent in cancer treatment in future.