Ayat B. Al-Ghafari

Antidepressants are cytotoxic to rat primary blood brain barrier endothelial cells at high therapeutic concentrations

Antidepressants are commonly employed for the treatment of major depressive disorders and other psychiatric conditions. We investigated the relatively acute cytotoxic effects of three commonly prescribed antidepressants: fluoxetine, sertraline, and clomipramine on rat primary blood brain barrier endothelial cells over a concentration range of 0.1-100μM. At therapeutic concentrations (0.1μM) no significant cytotoxicity was observed after 4, 24, or 48h. At high therapeutic to overdose concentrations (1-100μM), antidepressants reduced cell viability in proportion to their concentration and exposure duration. At 1μM, antidepressants significantly reduced mitochondrial membrane potential. At drug concentrations producing ~50% inhibition of cell viability, all drugs significantly reduced cellular oxygen consumption rates, activities of mitochondrial complexes I and III, and triggered a significant increase of lactate production. Fluoxetine (6.5μM) and clomipramine (5.5μM) also significantly lowered transcellular transport of albumin. The mechanism of cellular cytotoxicity was evaluated and at high concentrations all drugs significantly increased the production of reactive oxygen species, and significantly increased the activity of the pro-apoptotic caspases-3, 8, and 9. Comet assays revealed that all drugs were genotoxic. Pre-incubation of cells with glutathione significantly ameliorated antidepressant-induced cytotoxicity, indicating the potential benefit of treatment of overdosed patients with antioxidants.

The role of oxidative stress in antipsychotics induced ovarian toxicity

This study tested the hypothesis that oxidative stress could be an underlying mechanism for APs-induced ovarian cytotoxicity and reproductive dysfunction. Rat ovarian theca interstitial cells (TICs) were isolated and treated with four APs [chlorpromazine (CPZ), haloperidol (HAL), risperidone (RIS) and clozapine (CLZ)]. MTT assay was used to test the effects of these antipsychotics on TICs viability and to estimate their 50% inhibitory concentrations (IC50s). The effects of APs (IC50s and 1μM concentrations) on the activities of caspases-3, -8 and -9, reactive oxygen species (ROS) production, total intracellular glutathione and lipid peroxidation (LPO) in TICs were assessed. The effect of antioxidants (reduced glutathione (GSH) and quercetin) on the APs-induced cytotoxicity on TICs was investigated. MTT assay showed all APs to reduce TICs viability. CPZ, HAL and CLZ significantly increased the activity of caspases-3, -8 and -9 (P<0.0001, <0.0001 and <0.01, respectively). All APs at IC50s significantly (P<0.0001) increased ROS production, decreased total intracellular glutathione and increased LPO. MTT assay in the presence of antioxidants (reduced GSH (5mM) or quercetin (50mM)) showed each antioxidant to significantly inhibit the effects of APs at their IC50s on TICs viability. In conclusion, oxidative stress seems to be a possible mechanism for APs-induced ovarian and reproductive toxicity.

Effect of antipsychotics on mitochondrial bioenergetics of rat ovarian theca cells

BACKGROUND Antipsychotics (APs) are widely prescribed drugs, which are well known to cause reproductive adverse effects through mechanisms yet to be determined. The purpose of this study was to investigate the effect of antipsychotics on mitochondrial bioenergetics of rat ovarian theca cells as a possible mechanism of reproductive toxicity. METHODS Isolated rat theca interstitial cells (TICs) were treated with two typical (chlorpromazine [CPZ] and haloperidol [HAL]) and two atypical APs (risperidone [RIS] and clozapine [CLZ]). The effects of these APs on TICs bioenergetics (ATP content, mitochondrial complexes I and III activities, oxygen consumption rates (OCRs), mitochondrial membrane potential (MPP) and lactate production) and on steroidogenesis (androstenedione and progesterone synthesis) were investigated. RESULTS All APs resulted in a concentration-dependent decrease in the ATP content of TICs. All APs at their estimated IC50s (6μM, 21μM, 35μM and 37μM for CPZ, HAL, CLZ and RIS respectively) significantly decreased TICs OCRs (p<0.0001), MPP (p<0.0001) and significantly (p=0.0003) inhibited mitochondrial complex I activity. Only typical APs inhibited complex III (p=0.005). Also, APs at IC50s increased TICs lactate production to varying degrees. All APs used at their IC50s significantly inhibited progesterone (p=0.0022) and androstenedione (p=0.0027) production. Only CPZ was found to inhibit these hormones at the low concentration (1μM). CONCLUSION All four antipsychotics seem to inhibit mitochondrial bioenergetics and steroidogenesis in rats ovarian theca cells. These findings support the hypothesis that APs-induced reproductive toxicity may be through mechanisms involving mitochondrial insult>. Further research is required to establish the link between APs-induced mitochondrial dysfunction and disordered steroidogenesis.

Editor’s Highlight: Therapeutic Concentrations of Antidepressants Inhibit Pancreatic Beta-Cell Function via Mitochondrial Complex Inhibition

Diabetes mellitus risk is increased by prolonged usage of antidepressants (ADs). Although various mechanisms are suggested for their diabetogenic potential, whether a direct effect of ADs on pancreatic β-cells is involved is unclear. We examined this idea for 3 ADs: paroxetine, clomipramine and, with particular emphasis, fluoxetine, on insulin secretion, mitochondrial function, cellular bioenergetics, KATP channel activity, and caspase activity in murine and human cell-line models of pancreatic β-cells. Metabolic assays showed that these ADs decreased the redox, oxidative respiration, and energetic potential of β-cells in a time and concentration dependent manner, even at a concentration of 100 nM, well within the therapeutic window. These effects were related to inhibition of mitochondrial complex I and III. Consistent with impaired mitochondrial function, lactate output was increased and insulin secretion decreased. Neither fluoxetine, antimycin nor rotenone could reactivate KATP channel activity blocked by glucose unlike the mitochondrial uncoupler, FCCP. Chronic, but not acute, AD increased oxidative stress and activated caspases, 3, 8, and 9. A close agreement was found for the rates of oxidative respiration, lactate output and modulation of KATP channel activity in MIN6 cells with those of primary murine cells; data that supports MIN6 as a valid model to study beta-cell bioenergetics. To conclude, paroxetine, clomipramine and fluoxetine were all cytotoxic at therapeutic concentrations on pancreatic beta-cells; an action suggested to arise by inhibition of mitochondrial bioenergetics, oxidative stress and induction of apoptosis. These actions help explain the diabetogenic potential of these ADs in humans.

In Vitro Cytotoxic and Anticancer Effects of Zamzam Water in Human Lung Cancer (A594) Cell Line

BACKGROUND Zamzam water is naturally alkaline and rich in a variety of minerals which may represent a powerful tool for cancer therapy. In this research, the cytotoxic effects of Zamzam water were investigated in human lung cancer (A549) cell line and compared with human skin fibroblasts (HSF). METHODS Two different preparations of Zamzam water were used: Z1, with pH adjusted to 7.2 and Z2, with no pH adjustment. The effects of both treatments on the morphology of the A549 and HSF cell lines were investigated. The cell viability of HSF and A549 cells was identified by the MTT assay and trypan blue exclusion. Detection of apoptotic cells and cell cycle analyses were determined using flow cytometry. Moreover, reactive oxygen species (ROS) were measured for both cell lines. RESULTS Both Zamzam water treatments, Z1 and Z2 showed reductions in the cell viability of A549 cells. Cell death occurred via necrosis among cells treated with Z2. Cell cycle arrest occurred in the G0/G1 phases for cells treated with Z2. Cellular and mitochondrial ROS productions were not affected by either treatment. CONCLUSION Our findings indicate that Zamzam water might have potential therapeutic efficacy for lung cancer.

Implications of Isoprostanes and Matrix Metalloproteinase-7 Having Potential Role in the Development of Colorectal Cancer in Males

Background Colorectal cancer (CRC) is the third most common type of cancer and leading cause of death worldwide. Major risk factors involved in the development of CRC are increased dietary sources, genetics, and increasing age. Purpose of the study was to find the role of different variables in the progression of CRC. Methodology 50 blood samples from CRC patients and 20 samples from control were collected. Serum was separated from the blood by centrifugation. This serum was assessed for several antioxidants like superoxide dismutase (SOD), glutathione, glutathione peroxidase, glutathione reductase, catalase, vitamin A, C, and E, and pro-oxidants such as malondialdehyde, advanced oxidation protein products (AOPPs), and AGEs according to their respective protocols. Matrix metalloproteinase-7 (MMP-7) and isoprostanes were assessed by ELISA kits. Results Lower levels of GSH (4.86 ± 0.78 vs 9.65 ± 1.13 μg/dl), SOD (0.08 ± 0.012 vs 0.46 ± 0.017 μg/dl), CAT (2.45 ± 0.03 vs 4.22 ± 0.19 μmol/mol of protein), and GRx (5.16 ± 0.06 vs 7.23 ± 0.36 μmol/ml) in the diseased group were recorded as compared with control. Higher levels of GPx (6.64 ± 0.19 mmol/dl) were observed in the subjects in comparison with control group (1.58 ± 0.30 mmol/dl). Highly significant decreased levels of vitamin A (0.81 ± 0.07 vs 2.37 ± 0.15 mg/ml), vitamin E (15.42 ± 1.26 vs 25.96 ± 2.19 mg/ml), and vitamin C (47.67 ± 7.69 vs 80.37 ± 10.21 mg/ml) were observed in the patients in contrast to control group. The reversal of antioxidants in later stages of CRC may be due to compensatory mechanisms in cancerous cells. The levels of MDA (nmol/ml) were also assessed, which shows significantly increased level in CRC patients as compared with control groups (3.67 ± 0.19 vs 1.31 ± 0.27). The levels of protein oxidation products [AGEs (2.74 ± 0.16 vs 0.84 ± 0.05 IU) and AOPPs (1.32 ± 0.02 vs 0.82 ± 0.07 ng/ml)] were significantly increased in subjects as compared with control. The levels of MMP-7 (64.75 ± 3.03 vs 50.61 ± 4.09 ng/ml) and isoprostanes (0.71 ± 0.03 vs 0.16 ± 0.02 ng/ml) were also analyzed. This shows that the levels of isoprostanes increased due to high lipid peroxidation mediate higher levels of MMP-7, which promotes development of CRC. Conclusion Following study suggested that elevated oxidative and inflammatory status along with lipid peroxidation and matrix metalloproteinases are the chief contributors in the progression of CRC.

Contribution of Genetic Variants of ATP Transporters (ABCC1 and ABCG2) Genes With the Pathogenesis of Colorectal Cancer

1Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. 2Cancer Metabolism and Epigenetics Unit, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia. 3Experimental Biochemistry Unit, King Fahd Medical Research Centre, King Abdulaziz University, Jeddah, Saudi Arabia. 4Cancer and Mutagenesis Unit, King Fahd Medical Research Centre, King Abdulaziz University, Jeddah, Saudi Arabia.

ASSESSMENT OF THE ANTIOXIDANT PROPERTIES OF THE MOST COMMON COFFEE BREWS AVAILABLE IN THE LOCAL MARKETS OF THE WESTERN REGION OF SAUDI ARABIA

The purpose of the current study is to determine the antioxidant activity of the most commonly used coffee brews in the Western region of Saudi Arabia. Further, total phenolic and flavonoid contents were measured and antioxidants properties including ferric reducing antioxidant power and ferrous ion chelating activity, DPPH radical scavenging activity, and scavenging of hydrogen peroxide assays were also determined. Results of study revealed that phenolic contents were 741, 835 and 578 μg/ml of gallic acid/2 g of coffee, and the total flavonoid contents were 711, 802, and 828 μg/ml of catechin/2 g of coffee for Nescafé red mug, Turkish coffee and Espresso, respectively. The inhibition percentage of hydrogen peroxide showed highly significant reduction (p<0.001) in Turkish coffee compared to Nescafé red mug. Further, DPPH activity also showed highly significant reduction (p<0.01) in Espresso and Turkish coffee compared to Nescafé red mug, whereas, Espresso showed a highly significant increase (p<0.01) in reducing power activity. Regarding the metal chelating activity, Espresso showed a significant decrease (p<0.05) in metal chelation activity as compared to Nescafé red mug. The current study demonstrated that Nescafé red mug, Turkish coffee and Espresso, were the most consumed coffee types in the Western region of Saudi Arabia according to the survey. These three types of coffee showed high phenolic and flavonoid contents as well as a high antioxidant activity. Huda A. Al Doghaither * , Ashjan M. Almowalad, Ayat M. Shorbaji, Ayat B. Al-Ghafari, and Ulfat M. Omar

Association Of Telomerase Activity With The Prognosis Of Diabetes Mellitus In Saudi Population

Telomeres are important for chromosome stability and they are replenished by telomerase enzyme. Impaired telomerase activity has a strong association with ageing and diabetes. Research data have shown that telomerase activity plays a vital role in pathogenesis of type I and type II diabetes by influencing pancreatic cell regeneration. This study was aimed to assess telomerase activity in three diabetic groups (type I, type II and gestational diabetes mellitus (GDM)) and compare it with a population with normal blood glucose levels to establish a correlation between reduced telomerase activity and prognosis of diabetes in the Saudi population. This study was conducted on 200 Saudi diabetic and non-diabetic participants (20-60 years). Peripheral mononuclear cells (PMNCs) were extracted from the blood samples and the activity of telomerase in the PMNCs was detected by the TRAP-ELISA assay. Results of study revealed that there was no telomerase activity in control healthy group, moderate activity in type I, some activity in GDM, and significantly high activity in type II group. This might indicate that telomerase activity has a strong correlation to the prognosis of type II diabetes. However, the molecular mechanism still needs to be investigated. . KEYWORDS

The Effect of Adding Different Concentrations of Cows’ Milk on the Antioxidant Properties of Coffee

Polyphenols have been widely studied and considered as a health promoting and disease preventive agents in humans. Several studies investigated the antioxidant properties of polyphenols and their abilities to eliminate free radicals. In this study, the antioxidant activity of coffee in the presence of different types of cows’ milk at different concentrations, 10% or 20%, was investigated. Our results showed that milk could either enhance the scavenging of DPPH or decrease the metal chelating and metal reducing activity of polyphenol. However, more investigations are required to evaluate the mechanisms by which fats in milk can alter the antioxidant activity of coffee.