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AbstractMethotrexate (MTX) is an anti-folate chemotherapeutic commonly used to treat cancer and autoimmune diseases. Despite its widespread clinical use, MTX has been linked to serious neurotoxicity side effects. Vinpocetine (VNP) has been widely used clinically to treat many neurological conditions. This study was conducted to study the potential neuroprotective effects of VNP against MTX hippocampal intoxication in rats. Thirty-two rats were randomly allocated into 4 groups: (I) control (Vehicle); (II) VNP-treated group (20 mg/kg/day, p.o); (III) MTX-control (20 mg/kg/once, i.p.) group; and (IV) the VNP + MTX group. VNP was administered orally for 10 days, during which MTX was given intraperitoneally once at the end of day 5. Our data indicated that VNP administration significantly improved MTX-induced neuronal cell death, odema, vacuolation and degeneration. VNP attenuated oxidative injury mediated by significant upregulation of the Nrf2, HO-1, and GCLC genes, while the Keap-1 mRNA expression downregulated. Moreover, VNP suppressed cytokines release mediated by increasing IκB expression level while it caused a marked downregulation in NF-κB and AP-1 (C-FOS and C-JUN) levels. Additionally, VNP attenuated apoptosis by reducing hippocampal Bax levels while increasing Bcl2 levels in MTX-intoxicated rats. In conclusion, our results suggested that VNP significantly attenuated MTX hippocampal intoxication by regulating Keap-1/Nrf2, NF-κB/AP-1, and apoptosis signaling in these effects. [ABSTRACT FROM AUTHOR]

Background and Objectives: Diabetes mellitus (DM), is a metabolic disorder coupled with osteoporosis and elevated risk of bone fracture. This work aimed to clarify the impact of diabetes mellitus on bone remodeling changes and to study the role of vitamin D supplementation and mesenchymal stem cells (BM-MSCs) in decreasing bone remodeling changes in type II DM. Material and Methods: 30 adult male albino rats were classified into the control group (n-6) and the main experimental group (n=24). All rats in experimental group were given high fat diet and a single injection of streptozotocin (STZ) (45 mg/kg) intraperitoneally (IP) then after 8 weeks they were divided into; Type II DM group; continued without treatment, Type II DM +vitamin D group; received vitamin D orally daily, Type II DM +BM-MSCs group; received a once intravenous injection of 3 × 106 BM-MSCs and Combined treated group; treated with combined BM-MSCs and vitamin D. At the end of the experiment, Blood samples were drawn to measure glucose level, reactive Oxygen species (ROS), lipid profile (FFA), and tumor necrosis factor (TNF-α.). Femur bone was dissected for biochemical, histological, immunohistochemical, morphometric, and statistical analysis. Results: Mean serum levels of glucose, triglycerides, total cholesterol (TCH), ROS & TNF-α were significantly increased and associated with a significant decrease of HDL & FFA levels in Type II DM, Type II DM +vitamin D and Type II DM +BM-MSCs groups. In addition, the levels of Wnt/b-catenin and BCL2 were significantly decreased, while sclerostin and BAX, and the area percentage of VEGF were increased, in Type II DM, Type II DM +vitamin D and Type II DM +BMMSCs groups. Vitamin D and BM-MSCs encountered significant amelioration in biochemical markers and histological & immunohistochemical changes induced by STZ with a more obvious improvement in the combined treated group. Conclusion: Combined treatment of vitamin D and BM-MSCs significantly improved diabetic bone turnover compared to both vit. D, and MSCs injection alone by alleviating the antioxidative and anti-inflammatory effects. [ABSTRACT FROM AUTHOR]

Simple Summary: Endotoxin liver/kidney injury is characterized by oxidative stress, inflammation, and apoptosis that results in acute hepatic and kidney dysfunction with limited effective intervention. This study evaluated the potential protective efficacy of cepabiflas B and C (CBs) separated from Allium cepa against LPS (lipopolysaccharide)-induced hepatic and kidney damage and its possible mechanistic pathways. CBs effectively protected the liver and the kidney against LPS-induced damage. CBs ameliorated oxidative damage and enhanced Nrf2/HO-1 protective pathway. In reverse, CBs counteracted the activation of NF-κB/downstream cytokines which attenuated LPS-induced inflammatory response. CBs showed remarkable anti-apoptotic activity as it enhanced Bcl2 and suppressed Bax/caspase-3. Hence, this study elucidated the new therapeutic use of CBs in septic patients. Cepabiflas B and C (CBs) are flavonoid dimers separated from Allium cepa. They demonstrated antioxidant and α-glucosidase and protein tyrosine phosphatase 1B inhibition capacities. However, their anti-inflammatory activities and their effects on endotoxemia are unknown. The current study aimed at exploring the protective activities of CBs on lipopolysaccharide (LPS)-induced kidney and liver damage in mice and investigating the possible molecular mechanisms. Mice were orally treated with a low (40 mg/kg) or high (60 mg/kg) dose of CBs for five days prior to a single intraperitoneal injection of LPS (10 mg/kg). Samples of serum and hepatic and kidney tissues were collected 24 h after the LPS challenge. Changes in serum indices of hepatic and renal injury, pathological changes, molecular biological parameters, and proteins/genes related to inflammation and apoptosis of these organs were estimated. LPS injection resulted in deleterious injury to both organs as indicated by elevation of serum ALT, AST, creatinine, and BUN. The deteriorated histopathology of hepatic and renal tissues confirmed the biochemical indices. CBs treated groups showed a reduction in these parameters and improved histopathological injurious effects of LPS. LPS-induced hepatorenal injury was linked to elevated oxidative stress as indicated by high levels of MDA, 4-HNE, as well as repressed antioxidants (TAC, SOD, and GSH) in hepatic and kidney tissues. This was accompanied with suppressed Nrf2/HO-1 activity. Additionally, there was a remarkable inflammatory response in both organs as NF-κB signalling was activated and high levels of downstream cytokines were produced following the LPS challenge. Apoptotic changes were observed as the level and gene expression of Bax and caspase-3 were elevated along with declined level and gene expression of Bcl2. Interestingly, CBs reversed all these molecular and genetic changes and restricted oxidative inflammatory and apoptotic parameters after LPS-injection. Collectedly, our findings suggested the marked anti-inflammatory and anti-apoptotic activity of CBs which encouraged its use as a new candidate for septic patients. [ABSTRACT FROM AUTHOR]

Gastric ulcer is a serious medical condition that can be developed due to an imbalance in the protective and destructive factors of the gastric system. Available therapies do not provide definite cure, thus, there is an urge to seek for alternative treatments. Quercetin is a natural flavonoid that possesses antioxidant and anti-inflammatory properties. In the current study, the antiulcerogenic effect of quercetin in ethanol-induced gastric ulcer (EI-GU) rat model was compared to Antodine® (a reference drug), to elucidate the potential underlying mechanisms. Quercetin (50 mg/kg) and Antodine® (20 mg/kg) were given orally for one week post ulcer induction by ethanol. EI-GU was associated with downregulation of SOD, CAT, Nrf2 and HO1, and accompanied by upregulation of inflammatory markers (i.e., HMGB1, NF-κB and TNFα) and an increase in Bax/Bcl2 ratio. Administration of quercetin resulted in a significant reduction in gastric volume in the stomach of ulcerative rats by 86% and a significant decrease in gastric lesion count by 3.5- folds, as compared with the ulcerative rats. Moreover, rats treated with quercetin showed upregulation of Nrf2 by 3.3-fold change and in HO1 by 3.5-fold change when compared to ulcerated rats, and decreased HMGB1, TLR4, NF-κB p65 and TNF-α by 50%, 53%, 52.9% and 54.9%, respectively. Treatment of rats with quercetin reduced Bax and Bax/Bcl2 ratio and increased Bcl2 relative to ulcerated rats. Thus, it can be concluded that the ulcerogenic curative properties of quercetin were mediated by antioxidant, anti-inflammatory and antiapoptotic activities.

Objective: Diabetes is a metabolic disease which is linked to increased physical disabilities and muscle tissue damage. The aim of this study was to investigate the effect of interval and continued exercises with crocin on Bax/Bcl-2 ratio in diabetic obese rats. Materials and Methods: In this clinical trial study, 56 adult diabetic rats (high-fat diet and venous injection of streptozotocin) were selected and randomly assigned to groups (1), intense interval exercises (2), low intensity exercise (3), intense interval exercise with crocin consumption, (4) Low intensity exercise with crocin consumption, (5) Crocin consumption, (6) sham and (7) control were divided. Intense interval and low intensity exercise groups lasted for 8 weeks, three sessions per week, with intensity of 80 to 85 and 50 to 55 percent of maximum treadmill running, and crocin consumption groups for 8 weeks per day, mg / kg 25 crocin were taken peritoneal. To analyze the research hypotheses, Kolmogorov- Smirnov tests, independent T- tests and two-way multi-variable analysis of variance were used along with Benferron's comparison method. It should be noted that the significance level in all measurements was considered to be P-value≤ 0.05. Results: Results showed that exercise (P-value: 0.12) and crocin (P-value: 0.10) consumption had no significant effect on Bax/Bcl-2 gene expression in diabetic rats. Also interaction of exercise and crocin consumption on Bax/Bcl-2 was not significant (P-value: 0.12). Conclusion: It appears that exercise and crocin consumption have not interaction effect on improvement of Bax / Bcl-2 ratio in diabetic rats.

In 1990, pregabalin was introduced as a novel antiepileptic drug that acts by binding selectively to the alpha-2-delta subunits of voltage-gated calcium channels resulting in increasing neuronal GABA levels and inhibiting the release of exciting neurotransmitters. The aim of our study is to assess the hazardous effects of prolonged high-dose pregabalin (like that abused by addicts) on testes and to clarify the potential causative mechanisms. The current study was conducted on 70 adult male Wistar albino rats which were divided into 7 groups. In our study we evaluated the effect of pregabalin, at concentrations 150 and 300 mg/kg/day for 90 days, on hormones; FSH, LH, testosterone and prolactin secretion. Our study also evaluated the expression of apoptosis-related genes BAX and BCL2 in testicular tissue in addition to the western blotted analysis of p38 Mitogen activated protein kinases (p38 MAPK). The levels of reduced glutathione, malondialdehyde and superoxide dismutase were also measured. Pregabalin decreased testosterone level while FSH, LH and prolactin showed a significant increase. It also produced genotoxicity through reversal of the BAX/BCL2 ratio; increased p38 MAPK level and induction of oxidative stress markers. The concomitant administration of vitamin E significantly reduced all the previously mentioned biochemical and hormonal adverse effects caused by pregabalin. Pregabalin can adversely affect male fertility particularly in addicts and patients who are being treated with it for long periods as those suffering from neuropathies and seizures. Antioxidants like vitamin E could have a role in amelioration. [ABSTRACT FROM AUTHOR]

Introduction: Parkinson's disease (PD) is one of the most common neurodegenerative disorders. The neuroinflammation in the brain of PD patients is one of the critical processes in the immune pathogenesis of PD leading to the neural loss in the substantia nigra. Due to the anti-inflammatory effects of curcumin (CU) and low-level laser therapy (LLLT), we examined the protective effect of CU and LLLT on PC12 cells treated with 6-hydroxydopamine (6-OHDA) as a Parkinson model. Methods: PC12 cells were pretreated using various concentrations of 6-OHDA for 24 hours to induce oxidative and cellular damages. PC12-6-OHDA cells were co-treated with CU and LLLT. The effects of CU and LLLT on Bax/Bcl2 and LC3/ATG10 expression were analyzed by real-time PCR and cell viability was assessed by MTT assay. Cell A Software was used to calculate the length of the Neurite and cell body areas. Results: The results of this study show that the combination of CU dose-dependently and LLLT has a significant neuroprotective effect on cells and cellular death significantly decreases by increasing CU concentration. CU+LLLT decreases Bax/Bcl2 ratio which is an indicator of apoptosis and it also rescued a decrease in LC3 and ATG10 expression in comparison with 6-OHDA group. Conclusion: This study shows that the combination of 5 μM CU and LLLT has the best neuroprotective effect on PC12 cells against 6-OHDA by decreasing the BAX/BCL2 ratio. [ABSTRACT FROM AUTHOR]

Aims: In this study, novel synthesized 1,6-disubstituted-1-azacoumarin-3-carboxylic acid derivatives were designed, synthesized and evaluated as potential anticancer agents. Materials & methods: The cytotoxicity of novel 1-azacoumarin-3-carboxylic acid derivatives was tested using an MTT assay. High potency was shown by DNA flow cytometry on MCF-7 cells for compound 3b . In addition, topoisomerase IIβ, caspase 3/7, Bax and Bcl-2 enzymes were used to study apoptotic activity. In the same studies, molecular docking analysis assessed activity. Results & conclusion: Cytotoxicity screening identified multiple bioactive compounds, especially compound 3b . Analysis of DNA flow cytometry revealed that compound 3b exhibited cell cycle arrest. Compound 3b had an increase in the expression of Bax/Bcl-2 ratio and caspase 3/7, and a decrease in topoisomerase IIβ enzyme inhibition.

Objective: Diabetes is a metabolic disease which is linked to increased physical disabilities and muscle tissue damage. The aim of this study was to investigate the effect of interval and continued exercises with crocin on Bax/Bcl-2 ratio in diabetic obese rats. Materials and Methods: In this clinical trial study, 56 adult diabetic rats (high-fat diet and venous injection of streptozotocin) were selected and randomly assigned to groups (1), intense interval exercises (2), low intensity exercise (3), intense interval exercise with crocin consumption, (4) Low intensity exercise with crocin consumption, (5) Crocin consumption, (6) sham and (7) control were divided. Intense interval and low intensity exercise groups lasted for 8 weeks, three sessions per week, with intensity of 80 to 85 and 50 to 55 percent of maximum treadmill running, and crocin consumption groups for 8 weeks per day, mg / kg 25 crocin were taken peritoneal. To analyze the research hypotheses, Kolmogorov- Smirnov tests, independent T- tests and two-way multi-variable analysis of variance were used along with Benferron's comparison method. It should be noted that the significance level in all measurements was considered to be P-value≤ 0.05. Results: Results showed that exercise (P-value: 0.12) and crocin (Pvalue: 0.10) consumption had no significant effect on Bax/Bcl-2 gene expression in diabetic rats. Also interaction of exercise and crocin consumption on Bax/Bcl-2 was not significant (P-value: 0.12). Conclusion: It appears that exercise and crocin consumption have not interaction effect on improvement of Bax / Bcl-2 ratio in diabetic rats. [ABSTRACT FROM AUTHOR]

Objectives:Parkinson disease (PD) is a neurodegenerative disorder affecting mainly the motor system, as a result of death of dopaminergic neurons in the substantia nigra pars compacta. The present scenario of research in PD is directed to identify novel molecules that can be administered individually or co-administered with L-Dopa to prevent the L-Dopa-Induced Dyskinesia (LID) like states that arise during chronic L-Dopa administration. Hence, in this study, we investigated whetherMorinda citrifoliahas therapeutic effects in rotenone-induced Parkinson’s disease (PD) with special reference to mitochondrial dysfunction mediated intrinsic apoptosis. Methods:Male Sprague-Dawley rats were stereotaxically infused with rotenone (3 µg in both SNPc and VTA) and co-treated with the ethyl acetate extract ofMorinda citrifoliaand levodopa. Results:The results revealed that rotenone-induced cell death was reduced by MCE treatment as measured by decline in the levels of pro-apoptotic proteins. Moreover, MCE treatment significantly augmented the levels of anti-apoptotic Bcl2 and blocks the release of cytochrome c, thereby alleviating the rotenone-induced dopaminergic neuronal loss, as evidenced by tyrosine hydroxylase (TH) immunostaining in the striatum. Discussion:Taken together, the results suggest thatMorinda citrifoliamay be beneficial for the treatment of neurodegenerative diseases like PD. [ABSTRACT FROM AUTHOR]

Growing evidence has indicated that vitamin D (Vit D) regulates cell proliferation and differentiation in cancer cells. Accordingly, the present study was conducted to investigate the possible beneficial effects of Vit D on diethylnitrosamine (DEN)-induced liver preneoplasia. The effect of Vit D on HepG2 cells was investigated using MTT assay. Additionally, liver preneoplasia was induced in Swiss male albino mice by giving overnight fasted animals 5 consecutive doses of DEN (75 mg/kg/week). Oral treatment with Vit D (200 IU/kg/day) was initiated either 2 weeks before DEN (first protocol) or 1 week after the first dose of DEN injection (second protocol). At the end of the experiment, tissue levels of GGT, DPP-4, TNF-α, IL-6, CYP2E1, and CYP3A4 were also estimated. Moreover, the histopathological study of liver tissue and immunohistochemical detection of GST-P, PCNA, and NF-κB were performed. Vit D exerted a significant cytotoxic effect on HepG2 cells via significantly increasing BAX, p53, and BAX/Bcl2 ratio, and significantly decreasing Bcl2 mRNA expression. In both in vivo protocols, Vit D was capable of normalizing relative liver weight, PCNA, altered hepatocellular foci, and ductular proliferation. Moreover, Vit D significantly reduced the DEN-induced elevation of AST, ALT, ALP, GGT, DDP-4, TNF-α, IL-6, CYP2E1, liver DNA damage, GST-P, NF-κB, nuclear hyperchromasia/pleomorphism, cholestasis, and inflammatory cell aggregates, but significantly increased CYP3A4 content. In conculsion, current results reflect the potential impact of Vit D in the management of early stages of liver cancer. [Display omitted] • Vitamin D exerted a significant cytotoxic effect on HepG2 cells. • Vitamin D ameliorated DEN-induced liver preneoplasia in mice. • Vitamin D produced a significant reduction in tissue DPP-4 concentration • Vitamin D downregulated tissue CYP2E1 and upregulated tissue CYP3A4 activity. [ABSTRACT FROM AUTHOR]

Background: Curcuma purpurascens BI. (Zingiberaceae) commonly known as ‘Koneng Tinggang' and ‘Temu Tis' is a Javanese medicinal plant which has been used for numerous ailments and diseases in rural Javanese communities. In the present study, the apoptogenic activity of dichloromethane extract of Curcuma purpurascens BI. rhizome (DECPR) was investigated against HT-29 human colon cancer cells. Methods: Acute toxicity study of DECPR was performed in Sprague–Dawley rats. Compounds of DECPR were analyzed by the gas chromatography-mass spectrometry-time of flight (GC-MS-TOF) analysis. Cytotoxic effect of DECPR on HT-29 cells was analyzed by MTT and lactate dehydrogenase (LDH) assays. Effects of DECPR on reactive oxygen species (ROS) formation and mitochondrial-initiated events were investigated using a high content screening system. The activities of the caspases were also measured using a fluorometric assay. The quantitative PCR analysis was carried out to examine the gene expression of Bax, Bcl-2 and Bcl-xl proteins. Results: The in vivo acute toxicity study of DECPR on rats showed the safety of this extract at the highest dose of 5 g/kg. The GC-MS-TOF analysis of DECPR detected turmerone as the major compound in dichloromethane extract. IC50 value of DECPR towards HT-29 cells after 24 h treatment was found to be 7.79 ± 0.54 μg/mL. In addition, DECPR induced LDH release and ROS generation in HT-29 cells through a mechanism involving nuclear fragmentation and cytoskeletal rearrangement. The mitochondrial-initiated events, including collapse in mitochondrial membrane potential and cytochrome c leakage was also triggered by DECPR treatment. Initiator caspase-9 and executioner caspase-3 was dose-dependently activated by DECPR. The quantitative PCR analysis on the mRNA expression of Bcl-2 family of proteins showed a significant up-regulation of Bax associated with down-regulation in Bcl-2 and Bcl-xl mRNA expression. Conclusions: The findings presented in the current study showed that DECP suppressed the proliferation of HT-29 colon cancer cells and triggered the induction of apoptosis through mitochondrial-dependent pathway. [ABSTRACT FROM AUTHOR]

Highlights: [•] Diabetes increases the testicular FGF21 mRNA expression. [•] Deletion of FGF21 increases spontaneous and diabetes-induced testicular apoptosis. [•] Supplementation of exogenous FGF21 prevented diabetes-induced testicular apoptosis. [•] Prevention of FGF21 on apoptosis is related to reducing oxidative damage. [•] Deletion of FGF21 has no effect on testicular cell proliferation and inflammation. [Copyright &y& Elsevier]

Objective: Neem plant (Azadirachta indica) has been extensively used in Ayurvedic system of medicine for female fertility regulation for a long time, but its mechanism of action remains poorly understood. Hence, the present study was aimed to determine whether an increase of granulosa cell apoptosis is associated with aqueous neem leaf extract (NLE)-induced oocyte apoptosis. Materials and Methods: Sexually immature female rats of 20 days old were fed NLE (50 mg/day) for 10 days and then subjected to superovulation induction protocol. The morphological changes in cumulus oocyte complexes (COCs), rate of oocyte apoptosis, hydrogen peroxide (H2O2), total nitrite, and cytochrome c concentrations, inducible nitric oxide synthase (iNOS), cytochrome c, p53, Bcl2 and Bax expressions, deoxyribonucleic acid (DNA) fragmentation, and estradiol 17β level in granulosa cells collected from preovulatory COCs were analyzed. Results: Aqueous NLE increased H2O2 concentration and decreased catalase activity, increased iNOS expression and total nitrite concentration, increased p53, Bax, and p53 expressions but decreased Bcl2 expression, increased cytochrome c concentration and induced DNA fragmentation in granulosa cells. An increased granulosa cell apoptosis resulted in reduced estradiol 17β concentration and induced apoptosis in ovulated oocytes. Conclusion: We conclude that aqueous NLE-induced granulosa cell apoptosis through the mitochondria-mediated pathway, reduced estradiol 17β concentration and induced apoptosis in ovulated oocytes. Thus, granulosa cell apoptosis mediates NLE-induced oocyte apoptosis during female fertility regulation in rat. [ABSTRACT FROM AUTHOR]

To reveal the pathomechanisms of glaucoma, a common cause of blindness, suitable animal models are needed. As previously shown, retinal ganglion cell and optic nerve degeneration occur in βB1-CTGF mice. Here, we aimed to determine possible apoptotic mechanisms and degeneration of different retinal cells. Hence, retinae were processed for immunohistology (n = 5–9/group) and quantitative real-time PCR analysis (n = 5–7/group) in 5- and 10-week-old βB1-CTGF and wildtype controls. We noted significantly more cleaved caspase 3+ cells in βB1-CTGF retinae at 5 (p = 0.005) and 10 weeks (p = 0.02), and a significant upregulation of Casp3 and Bax/Bcl2 mRNA levels (p < 0.05). Furthermore, more terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL+) cells were detected in transgenic mice at 5 (p = 0.03) and 10 weeks (p = 0.02). Neurofilament H staining (p = 0.01) as well as Nefh (p = 0.02) and Tubb3 (p = 0.009) mRNA levels were significantly decreased at 10 weeks. GABAergic synapse intensity was lower at 5 weeks, while no alterations were noted at 10 weeks. The glutamatergic synapse intensity was decreased at 5 (p = 0.007) and 10 weeks (p = 0.01). No changes were observed for bipolar cells, photoreceptors, and macroglia. We conclude that apoptotic processes and synapse loss precede neuronal death in this model. This slow progression rate makes the βB1-CTGF mice a suitable model to study primary open-angle glaucoma. [ABSTRACT FROM AUTHOR]

Astaxanthin (AST) is related to apoptosis but the details of the mechanism of how AST makes apoptosis is not clear. The present study investigated apoptotic effects of AST to SKBR3, a breast cancer cell line in detail. Cell viability assay showed cellular proliferation and morphological changes of the cells were observed under AST treatment. FACS analysis indicated that AST blocked cell cycle progression at G0/G1, suppressed proliferation dose-dependently, and induced apoptosis of the cells. The apoptosis of the cells by AST was further demonstrated through the decreased expression level of mutp53 and cleaved a PARP-1 fragment, respectively. In addition, AST induced the intrinsic apoptosis of the cells by activation of Bax/Bcl2, cleaved caspase-3, and cleaved caspase-9 as well as the phosphorylation of ERK1/2, JNK, and p38. Furthermore, AST decreased production of intracellular reactive oxygen species as well as modulated expressions of superoxide dismutases and Pontin, an anti-apoptotic factor. Co-immunoprecipitation assay revealed AST reduced interaction between Pontin and mutant p53. Taken together, these studies proved that AST regulates the expression of apoptotic molecules to induce intrinsic apoptosis of the cells, suggesting AST therapy might provide an alternative for improving the efficacies of other anti-cancer therapies for breast cancer. [ABSTRACT FROM AUTHOR]

Elimination of senescent cells (SnC) is anti-atherogenic, but the specific contribution of senescent vascular endothelial cells (EC) is unknown. We inactivated angiopoietin like-2 (angptl2), a marker of SnEC and a pro-atherogenic cytokine in LDLr -/- , hApoB 100 +/+ atherosclerotic (ATX) mice. Three months after a single vascular delivery of a small hairpin (sh)Angptl2 in 3-month old ATX mice using an adeno-associated virus serotype 1 (AAV1), aortic atheroma plaque progression was slowed by 58% (p<0.0001). In the native aortic endothelium, angptl2 expression was decreased by 80%, in association with a reduced expression of p21 , a cyclin-dependent kinase inhibitor overexpressed in growth-arrested SnC. Endothelial activation was reduced (lower Icam-1, Il-1β and Mcp-1 expression), decreasing monocyte Cd68 expression in the endothelium. One week post-injection, the ratio Bax/Bcl2 increased in the endothelium only, suggesting that angptl2 + /p21 + SnEC were eliminated by apoptosis. Four weeks post-injection, the endothelial progenitor marker Cd34 increased, suggesting endothelial repair. In arteries of atherosclerotic patients, we observed a strong correlation between p21 and ANGPTL2 (r=0.727, p=0.0002) confirming the clinical significance of angptl2 -associated senescence. Our data suggest that therapeutic down-regulation of vascular angptl2 leads to the clearance of SnEC by apoptosis, stimulates endothelial repair and reduces atherosclerosis.

Arsenic (As), cadmium (Cd), and lead (Pb) in select combinations are proved to affect the viability of astrocyte. However, their role in glioma, an aggressive astroglial tumor, is unexplored. We analyzed the effect of As+Cd+Pb on C6-glioma cells derived from rat glioma. We determined the lethal concentration (LC) of individual metal, and then treated C6-glioma cells with As+Cd+Pb at LC-5 (As: 5 mM, Cd: 2.5 mM and Pb: 15 mM), and concentrations that were double or triple of LC5. As+Cd+Pb induced dose-dependent reduction in C6-glioma viability. Cell death was due to apoptotic DNA fragmentation, detected through terminal deoxynucleotidyl transferase-mediated dUTP-nick-end labeling. An enhanced cleavage of caspase-9 indicated the apoptosis to be mitochondria-mediated. An increase in pro-apoptotic Bcl-2-associated-X protein (Bax) and decrease in anti-apoptotic Bcl2 resulting in a Bax/Bcl2 ratio > 1.0 validated mitochondrial apoptosis. Exploring apoptotic regulatory mechanism revealed an alteration in glial cell morphology and augmentation of astroglial marker, glial fibrillary acidic protein (GFAP), that demonstrated co-localization with cleaved caspase-9. The glial activation was accompanied by inflammation, involving the up-regulation of interleukin-1 (IL-1) and IL-1-receptor. IL-1 also contributed to apoptosis, as evident from the attenuation of cleaved caspase-9 upon treatment with IL-1receptor antagonist. Investigating the involvement of Mitogen-activated protein kinases (MAPKs) revealed the activation of P38 as indicated by an increased phospho-p38 expression. p38-MAPK inhibitor, SB203580, prevented caspase-9 activation, which further suppoted the involvement of p38-MAPK in C6-glioma apoptosis. Overall our data demonstrate the toxic effect of As+Cd+Pb on C6-glioma, which is mediated by mitochondria-dependent apoptosis that requires astroglial activation, inflammation and p38-MAPK signaling. As+Cd+Pb combination treatment may have a potential therapeutic usage against glial tumors.

Myricetin is a flavonol found in various berries, herbs, and walnuts. Previous studies have demonstrated that myricetin has anticancer effects against several types of cancer, including hepatocarcinoma, skin carcinoma, and pancreatic cancer. However, the anticancer activity of myricetin on human colon cancer has not been yet established. In the present study, we investigated the anticancer effects of myricetin on HCT-15 human colon cancer cells. We found that myricetin induces cytotoxicity and DNA condensation in human colon cancer cells in a dose-dependent manner. We also determined that myricetin increases the BCL2-associated X protein/B-cell lymphoma 2 ratio, but not cleavage of caspase-3 and -9. In addition, myricetin induced the release of apoptosis-inducing factor from mitochondria. These results suggest that myricetin induces apoptosis of HCT-15 human colon cancer cells and may prove useful in the development of therapeutic agents for human colon cancer.