Gallic acid potentiates the apoptotic effect of paclitaxel and carboplatin via overexpression of Bax and P53 on the MCF-7 human breast cancer cell line.

Despite advances in treatment, breast cancer remains the widest spread disease among females with a high mortality rate. We investigated the potential effects of gallic acid (GA) as supportive therapy in the management of breast cancer. Anti-cancer activity with GA alone or in combination with paclitaxel and/or carboplatin was assessed by MTT assay and flow cytometry using annexin V/propidium iodide. The mechanism underlying the antiproliferative effects was investigated by measuring the expression of the pro-apoptotic marker (Bax), CASP-3, anti-apoptotic (Bcl-2), and, tumor suppressor (p53) by real-time polymerase chain reaction (RT-PCR) and western blot analysis. Cell cycle analysis was performed for the MCF-7 breast cancer cell line. GA, paclitaxel, and carboplatin alone or in combination arrested cell cycle progression at the G2/M phase and induced Pre-G1 apoptosis. RT-PCR showed that the triplet combination significantly raised P53, Bax, and CASP-3 mRNA expression (20.1 ± 1.41, 16.6 ± 0.43, and 20.04 ± 1.61, respectively) in MCF-7 cells when compared to single or combined treatment (p < .0001) while anti-apoptotic Bcl-2 mRNA levels were decreased in all treated groups compared to untreated cells. Western blot data of tested apoptotic factors were consistent with RT-PCR results. For the first time, we show that a minimum non-toxic concentration of GA increased the efficacy of paclitaxel- and carboplatin-induced MCF-7 apoptotic cell death.

IL-6 and NFE2L2: A putative role for the hepatoprotective effect of N. Sativa, P. Ginseng and C. Sempervirens in AFB-1 induced hepatocellular carcinoma in rats.

In this study, we investigated possible hepato-protective effects of N. Sativa, P. Ginseng, and C. Sempervirens in Aflatoxin B1 (AFB-1) induced hepatocellular carcinoma rat model. Fifty-four male albino rats were randomly assigned to experimental groups. Alcoholic extracts of aforementioned herbs were administered orally for 28 days at different doses. IL-6, hs-CRP, MDA, SOD and NFE2L2 were determined using ELISA. Histopathological changes in treated groups were examined. Herbal treatment significantly reduced IL-6, hs-CRP, and MDA (P < 0.001) whereas it significantly increased SOD (p < 0.001). C. Sempervirens 600 and N. Sativa 1000 increased NFE2L2 level compared to P. Ginseng 500 group (P value<0.01). Histopathological evaluation of treated groups showed different grades of healing of the liver. This study confirms a beneficial hepatoprotective effect for aforementioned herbal extracts orally administered in rat model of AFB1 induced HCC. This effect is putatively mediated via modulation of inflammatory cytokines as well as amelioration of oxidative stress.

A putative Chondroprotective role for IL-1ß and MPO in herbal treatment of experimental osteoarthritis.

BACKGROUND: Herbal treatment may have a chondroprotective and therapeutic effect on Osteoarthritis (OA). We investigated the mechanism of action of ginger and curcumin rhizomes cultivated in Egypt in treatment of OA in rat model. METHODS: Thirty-five albino rats were intra-articularly injected with Monosodium Iodoacetate in the knee joint. Ginger and curcumin was orally administered at doses of 200 and 400 mg/kg (F200 and F400). Serum levels of cartilage oligomeric matrix protein (COMP), hyaluronic acid (HA), malondialdehyde (MDA), myeloperoxidase (MPO), Interleukin-1 beta (IL-1ß) and superoxide dismutase activity (SOD) were measured using ELISA. The composition of the herbal formula hydro-ethanolic extract was characterized using UPLC-ESI-MS. Histopathological changes in injected joints was examined using routine histopathology. Statistical analysis was performed using one-way ANOVA. RESULTS: Serum levels of COMP, HA, MPO, MDA, and IL-1ß were significantly decreased in F 200, F 400 and V groups when compared to OA group (P value <0.0001). On the other hand SOD levels were significantly elevated in treated groups compared to OA groups (P value <0.0001). CONCLUSIONS: The ginger/curcumin at 1:1 had chondroprotective effect via anti-inflammatory and antioxidant effect in rat OA model. Further pharmacological and clinical studies are needed to evaluate this effect.

IL-10 and TGF-ß: Roles in chondroprotective effects of Glucosamine in experimental Osteoarthritis?

OBJECTIVE: Osteoarthritis (OA) is a complex disease of the whole joint. Glucosamine (GlcN) treatment may have a chondroprotective effect on OA. We investigated the mechanism of action of glucosamine treatment through interleukin-10 (Il-10) and transforming growth factor ß-1 (TGF ß-1). METHODS: Thirty male albino rats were used. A single intraarticular (i.a.) injection of 2mg of Monosodium Iodoacetate (MIA) was injected into the knee joint of anesthetized rats. GlcN (50 or 100mg/kg/day, p.o. for 2 month) was administered orally. Serum levels of Il-10 and TGF-ß1 were determined by ELISA. Histopathological changes in treated and control joints were examined using hematoxylin-eosin (H & E) staining. RESULTS: The mean serum level of IL-10 significantly decreased in the OA group compared to control group (P value<0.0001). On the other hand, mean serum level of IL-10 significantly increased in GlcN treated groups when compared to the OA group (P value<0.0001). Serum level of TGF ß-1 was significantly elevated in OA group compared to control group (P value<0.0001). On the other hand, the mean serum level of TGF ß-1 was significantly decreased in the GlcN treated groups when compared to the OA group (P value<0.0001). Histopathological evaluation of GlcN treated groups showed different grades of healing, according to Osteoarthritis Research Society International (OARSI) grading system. CONCLUSION: Our results showed that IL-10 and TGF-ß1 possibly mediate GlcN chondroprotective effects in OA. Both serum biomarkers can be useful in the follow-up of articular cartilage damage in clinical settings.

Resistin mediates tomato and broccoli extract effects on glucose homeostasis in high fat diet-induced obesity in rats.

BACKGROUND: Resistin is an adipocyte hormone that regulates glucose metabolism. Elevated levels of resistin may cause insulin resistance. This may link obesity, and increased fat mass to type II diabetes and insulin resistance. We hypothesized that treatment with tomato and broccoli extracts regulates glucose homeostasis via modulation of resistin levels in high fat diet-induced obesity rats (HFD). METHODS: Forty-eight male albino rats were divided into 8 groups as follows: control, HFD, stop fat diet (SD), Tomato 200 mg/kg (T200), Tomato 400 mg/kg (T400), Broccoli 200 mg/kg (B200), Broccoli 400 mg/kg (B400), and Chromax (CX). Treatment continued for 1 month. Serum levels of resistin, leptin, adiponectin, glucose and insulin were measured using ELISA and spectrophotometry. RESULTS: Serum levels of resistin were significantly reduced in the T 200, T 400, B 200, B 400 and CX groups to: 4.13 ± 0.22 ng/ml, 1.51 ± 0.04 ng/ml, 4.13 ± 0.22 ng/ml, 2.32 ± 0.15 ng/ml and 1.37 ± 0.03 ng/ml, respectively, compared to HFD group and SD group (P value < 0.0001). Non-significant differences were found between T 400, B 400 and CX groups. Serum levels of leptin were significantly reduced in the T 400 (22.7 ± 0.84 pg/ml) group compared to the B 400 (41 ± 2.45 Pg/ml) and CX groups (45.7 ± 2.91 Pg/ml), P value < 0.001. Serum levels of adiponectin were significantly increased in the T 400 group (131 ± 3.84 pg/ml) compared to the CX group (112 ± 4.77 pg/ml), P value < 0.01. CONCLUSIONS: Our results demonstrate that tomato and broccoli extract treatment regulates glucose homeostasis via reduction of serum resistin and may be a useful non-pharmacological therapy for obesity.

Circadian Pattern of Melatonin MT1 and MT2 Receptor Localization in the Rat Suprachiasmatic Nucleus.

The suprachiasmatic nucleus (SCN) is the master circadian pacemaker. The pineal hormone melatonin is involved in the regulation of circadian phase. As a part of the circadian system, its synthesis and secretion is under SCN control. On the other hand, melatonin feeds back on the SCN to regulate its function. Melatonin has two specific windows of time at which it regulates SCN function, namely dusk and dawn. It has been suggested that melatonin exerts its effect on the SCN during that specific window of time via one or both of its specific receptors, MT1 or MT2. The hypothesis that the density of these receptors varies across the circadian cycle was tested. Using immunohistochemistry with receptor-specific antibodies, the localization and distribution of melatonin receptors MT1 and MT2 was studied in the SCN at different Zeitgeber times (ZT): ZT 11-13 (dusk), 23-01 (dawn), 5-7 (mid-day), and 17-19 (midnight). Our results show that MT1 receptor density significantly increased at dusk relative to dawn and midnight (p<0.01 and p<0.001 respectively). Although MT1 receptors were widespread in the SCN and parts of the optic chiasm at dusk, they were restricted to the SCN during the mid-day period. MT2 receptors were not detected in the SCN. Thus, we find that melatonin receptor MT1 density and distribution varies with circadian time. This creates a time window during which melatonin can affect the operation of the SCN. We also find that melatonin regulates SCN function via MT1 receptors with a minimal role for MT2.