Gac fruit extracts ameliorate proliferation and modulate angiogenic markers of human retinal pigment epithelial cells under high glucose conditions

Objective: To investigate the impact of the extracts of Gac fruit parts(peel, pulp, seed, and aril) on the cell viability and angiogenesis markers of human retinal pigment epithelial(ARPE-19) cells under high glucose conditions. Methods: The effect of the extracts of Gac fruit peel, pulp, seed and aril on the ARPE-19 cells was determined using MTT viability assay, Trypan blue dye and morphological changes were observed using light microscopy. Enzyme-linked immunosorbent-based assay was performed to evaluate the effect of Gac fruit parts on the reactive oxygen species(ROS), vascular endothelial growth factor(VEGF) and pigmented epithelium-derived factor(PEDF) secretions. Results: High glucose(HG) at 30 mmol/L increased ARPE-19 cell viability and ROS and VEGF secretions. While, the exposure of ARPE-19 cells in high glucose condition to Gac fruit extracts led to inhibition of cell viability, induced morphological changes, decreased ROS and VEGF secretions, and increased PEDF level. Gac pulp, seed, and aril at 1 000 μg/mL showed significant inhibition activities [(7.5 ± 5.1)%,(2.7 ± 0.5)%,(3.2 ± 1.1)%, respectively] against HG-induced ARPE-19 cell viability. The findings also demonstrated that Gac aril at 250 μg/mL significantly decreased ROS and VEGF levels [(40.6 ± 3.3) pg/mL,(107.4 ± 48.3) pg/mL, respectively] compared to ROS [(71.7 ± 2.9) pg/mL ] and VEGF [(606.9 ± 81.1) pg/mL] in HG untreated cells. Moreover, 250 μg/mL of Gac peel dramatically increased PEDF level [(18.2 ± 0.3) ng/mL] compared to that in HG untreated cells [(0.48 ± 0.39) ng/mL]. Conclusions: This study indicates that the extracts of Gac peel, pulp, seed and aril reduced cell viability, minimized ROS generations and showed angiogenic activities. Therefore, our findings open new insights into the potentiality of Gac fruit against HG-related diabetic retinopathy disease.

Anti-breast cancer properties and toxicity of Dillenia suffruticosa root aqueous extract in BALB/c mice

Objective:To determine the anti-breast cancer activities and the safety oral consumption of Dillenia suffruticosa root aqueous extract(DRAE)in BALB/c mice.Methods:In the anti-breast cancer study,female BALB/c mice were divided into five groups(n=12),which were(1)positive control(with breast cancer,untreated),(2)negative control(without breast cancer,untreated)and other three groups of mice with breast cancer treated with 1 000,500 and 250 mg/kg of DRAE,respectively,by oral gavage for 28 days.All mice except from the negative control group were injected into the mammary fat pad with 4T1 cells(1×1054T1 cells/0.1 m L of phosphate buffer solution).DRAE was administered orally on Day 11 after the tumor has developed.Results:The tumor volume of the 1 000 mg/kg of DRAE group reduced significantly compared to the positive control while treatment with 500 mg/kg of DRAE had significantly inhibited metastasis to the heart.In the acute toxicity study,treatment with up to5 000 mg/kg of DRAE was not toxic to the animals,indicating its safety when a large amount of this plant extract was ingested.Based on the sub-acute toxicity study,treatment of the highest dose of DRAE(1 000 mg/kg)had mild liver toxicity indicated by mild focal hemorrhage.Conclusions:DRAE possesses anti-breast cancer properties but at the same time it shows mild toxicity to the liver.The non observable adverse effect dose for DRAE is500 mg/kg.

Antioxidant compounds and capacities of Gac (Momordica cochinchinensis Spreng) fruits

Objective: To identify and determine the composition of antioxidant compounds, and to evaluate the antioxidant abilities of Gac fruit parts(peel, pulp, seed and aril) grown in Malaysia. Methods: LC-MS/MS was used for identification of antioxidant compounds and UV-Vis for estimation of the contents of phenolics, flavonoids, and carotenoids. Lycopene and β-carotene were quantified using high-performance liquid chromatography. DPPH(2, 2-diphenyl-1-picrylhydrazyl) and ferric reducing antioxidant power assays were employed to evaluate antioxidant capacities. Results: Phytochemicals were found amongst all the fruit parts. Notably, significant amounts of carotenoids [(107.4 ± 4.5),(85.7 ± 4.4),(110.6 ± 2.1) mg/100 g dry weight(DW)], and relatively high levels of both phenolics [(27.3 ± 1.7),(28.9 ± 2.4),(30.8 ± 2.7) mg/100 g DW] and flavonoids [(38.1 ± 2.2),(8.8 ± 1.3),(24.5 ± 3.3) mg/100 g DW] were found in the fruit's peel, pulp and aril, respectively. Seed part also showed a relatively high level of flavonoids [(18.1 ± 2.3) mg/100 g DW]. Lycopene and β-carotene were found to be significantly high(P < 0.05) in aril [(579.3 ± 22.7) and(621.0 ± 35.0) μg/g DW], followed by peel [(51.0 ± 7.5) and(210.0 ± 12.5) μg/g DW] and pulp [(37.6 ± 10.9) and(205.6 ± 22.1) μg/g DW)]. Antioxidant assays revealed that aril possessed the highest scavenging activity(IC_(50) = 865 μg/mL), while the peel possessed the highest ferric reducing power of 140 μmol FeSO4/μg. Conclusions: The current results demonstrate that Gac fruit grown in Malaysia is a rich source of phytochemicals, especially carotenoids, and possesses antioxidant activities. Thus, such findings suggest Gac fruit as a source of an antioxidant plant.

Phenolics-saponins rich fraction of defatted kenaf seed meal exhibits cytotoxicity towards cancer cell lines

Objectives: To determine the cytotoxicity of crude ethanolic extract, n-butanol fraction and aqueous fraction on selected cancer cell lines, and to observe the morphological changes of the cancer cells treated with n-butanol fraction.Methods: The cytotoxic effect of n-butanol fraction, crude ethanolic extract and aqueous fraction on breast cancer(MCF-7 and MDA-MB-231), colon cancer(HT29), lung cancer(A549), cervical cancer(He La) and normal mouse fibroblast(3T3) cell lines was determined using MTT assay. The morphological changes of the treated cells were observed under an inverted light microscope.Results: n-Butanol fraction was the most cytotoxic towards HT29 and MCF-7 cells in a dose-dependent manner compared to crude ethanolic extract and aqueous fraction(P < 0.05). The IC50 of n-butanol fraction for HT29 and MCF-7 was(780.00 ± 28.28)and(895.00 ± 7.07) mg/m L, respectively. Cell shrinkage, membrane blebbing and formation of apoptotic bodies were noted following treatment of HT29 cells with n-butanol fraction.Conclusions: In conclusion, n-butanol fraction was more cytotoxic than crude ethanolic extract and aqueous fraction towards the selected cancerous cell lines and induced apoptosis in HT29 cells.

Brewers' rice modulates oxidative stress in azoxymethanemediated colon carcinogenesis in rats

AIM: To investigate the mechanistic action of brewers' rice in regulating the Wnt/nuclear factor-kappa B(NF-κB)/Nrf2-signaling pathways during colon carcinogenesis in male Sprague-Dawley rats.METHODS: Male Sprague-Dawley rats were randomly divided into the following five groups(six rats in each group):(G1) normal,(G2) azoxymethane(AOM) alone,(G3) AOM + 10%(weight(w)/weight(w)) brewers' rice,(G4) AOM + 20%(w/w) brewers' rice, and(G5) AOM + 40%(w/w) brewers' rice. They were intraperitoneally administered 15 mg/kg body weight of AOM in saline once weekly over a twoweek period and treated with an American Institute of Nutrition(AIN)-93 G diet containing 10%, 20%, and 40%(w/w) brewers' rice. The m RNA levels of glycogen synthase kinase 3β(GSK 3β), β-catenin, key inflammation markers, nuclear factor E2-related factor 2(Nrf2), and heme oxygenase-1(HO-1)-dependent transcriptional activity were assessed by quantitative real-time polymerase chain reaction analyses. The colon superoxide dismutase, malondialdehyde, and nitric oxide levels were also analyzed to assess the antioxidant effect of these treatments. The results were analyzed using one-way analysis of variance(ANOVA), and a P value of < 0.05 was considered significant.RESULTS: The overall analyses demonstrated that the dietary administration of brewers' rice in AOM-induced rat colon carcinogenesis resulted in the transcriptional upregulation of GSK 3β, inducible nitric oxide synthase(i NOS), Nrf2, and HO-1. We discovered that the dietary administration of brewers' rice downregulated the β-catenin and NF-κB m RNA levels. A significant reduction in β-catenin expression was found in the groups administered with 20%(0.611 ± 0.034) and 40%(0.436 ± 0.045)(w/w) brewers' rice compared with that of the group treated with AOM alone(1.000 ± 0.064)(P < 0.05). The NF-κB expression was significantly lower between the AOM-alone group(1.000 ± 0.048) and those groups fed with diets containing 10%(w/w) brewers' rice(0.255 ± 0.022), 20%(w/w) brewers' rice(0.450 ± 0.045), or 40%(w/w) brewers' rice(0.541 ± 0.027)(P < 0.05). Brewers' rice improved the antioxidant levels, indicating that brewers' rice can enhance effective recovery from oxidative stress induced by AOM.CONCLUSION: Our results provide evidence that brewers' rice can suppress colon cancer via the regulation of Nrf2 expression and the inhibition of the Wnt/NF-κB signaling pathways.

Edible Bird's Nest Attenuates Menopause-Related Bone Degeneration in Rats via Increasing Bone Estrogen-Receptor Expression

Objective:To investigate the mechanistic basis for the attenuation of bone degeneration by edible bird's nest(EBN)in ovariectomized rats.Methods:Forty-two female Sprage-Dawley rats were randomized into 7 groups(6 in each group).The ovariectomized(OVX)and OVX+6%,3%,and 1.5%EBN and OVX+estrogen groups were given standard rat chow alone,standard rat chow+6%,3%,and 1.5%EBN,or standard rat chow+estrogen therapy(0.2mg/kg per day),respectively.The sham-operation group was surgically opened without removing the ovaries.The control group did not have any surgical intervention.After 12 weeks of intervention,blood samples were taken for serum estrogen,osteocalcin,and osteoprotegerin,as well as the measurement of magnesium,calcium abd zinc concentrations.While femurs were removed from the surrounding muscles to measure bone mass density using the X-ray edge detection technique,then collected for histology and estrogen receptor(ER)immunohistochemistry.Results:Ovariectomy altered serum estrogen levels resulting in increased food intake and weight gain,while estrogen and EBN supplementation attenuated these changes.Ovariectomy also reduced bone ER expression and density,and the production of osteopcalcin and osteorotegerin,which are important pro-osteoplastic hormones that promote bone mineraliztion and density.Conversely,estrogen and EBN increased serum estrogen levels leading to increased bone ER expression,pro-osteoplastic hormone production and bone density(all P<0.05).Conclusion:EBN could be used as a safe alternative to hormone replacement therapys for managing menopausal complications like bone degeneration.

AIM associated with the IgM pentamer: attackers on stand-by at aircraft carrier

Circulating immunoglobulin M(IgM)exists in a pentameric form,possessing a polyreactive nature that responds not only to foreign antigens but also to autoantigens;thus,it is involved in both beneficial and detrimental immune responses,including protection from infection and the progression of autoimmunity.On the other hand,IgM also behaves as a carrier of the apoptosis inhibitor of macrophage(AIM)protein,storing a large amount of the inactivated form of AIM in the blood through this association.Under different disease conditions,AIM can dissociate from IgM locally or systemically to exert its function,inducing the removal of various biological debris such as excess fat,bacteria,cancer cells or dead cell debris.Most typically,upon induction of acute kidney injury(AKI),IgM-free AIM is filtered by the glomerulus in the kidney,which stimulates the clearance of intraluminal dead cells debris at the obstructed proximal tubules,thereby facilitating the repair of kidney injury.Interestingly,cats exhibit a deficiency in AIM release from IgM,which may increase their susceptibility to renal failure.Conversely,association with AIM inhibits IgM binding to the Fcα/μreceptor on follicular dendritic cells at the splenic germinal center,thereby protecting the IgM immune complex from Fcα/μreceptor-mediated internalization,which supports IgM-dependent antigen presentation to B cells and stimulates high-affinity IgG antibody production.The regulation of AIM–IgM binding,resulting from the discovery of reciprocal actions between AIM and IgM,could lead to the development of novel therapies against different diseases.