Protective effect of astragalus-containing serum on cobalt chloride-induced hypoxia injury in ARPE-19 cells / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the protective effect of astragalus-containing serum on cobalt chloride(CoCl2)-induced hypoxia injury of human retinal pigment epithelial cells(ARPE-19), so as to explore whether astragalus can improve diabetic retinopathy(DR)by anti-oxidative stress.<p>METHODS: The ARPE-19 hypoxia model induced by CoCl2 was established and divided into the following 5 groups: normal group(cells were cultured normally without any treatment), hypoxia model group(200μmol/L CoCl2), blank serum group(200μmol/L CoCl2+blank serum), low-dose drug-containing serum group(200μmol/L CoCl2+10% medicated serum)and high-dose drug-containing serum group(200μmol/L CoCl2+20% medicated serum); CCK-8 detects cell viability; Detect the levels of reduced glutathione(GSH)and malondialdehyde(MDA)in the cell supernatant with a kit; ELISA was used to detect the content of hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)in cell culture medium; Real-time quantitative PCR(qPCR)to detect the mRNA levels of VEGF, HIF-1α and Prolyl hydroxylase-2(PHD-2); The expressions of VEGF, HIF-1α and PHD-2 were detected by Western Blot.<p>RESULTS: Hypoxia model of ARPE-19 can successfully establish by CoCl2 at 200μmol/L. Low-dose and high-dose astragalus-containing serum could inhibit hypoxia-induced ARPE-19 proliferation(<i>P</i><0.05), increase the GSH level and reduce the MDA content in ARPE-19 with hypoxic injury(<i>P</i><0.05). Low-dose and high-dose astragalus-containing serum could inhibit the expression of HIF-1α and VEGF in ARPE-19 hypoxic injury supernatant(<i>P</i><0.05), as well as the mRNA and protein expressions of VEGF, HIF-1α and PHD-2 in ARPE-19(<i>P</i><0.05).<p>CONCLUSION: Low-dose and high-dose astragalus-containing serum alleviates the hypoxia injury of ARPE-19 induced by CoCl2 through anti-oxidant effect.

Study on the apoptosis of human retinal pigment epithelium cells induced by EGCG / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the effect of epigallocatechin gallate(EGCG)on the apoptosis of human retinal pigment epithelium(ARPE-19)cells and its mechanism. METHODS:The ARPE-19 cells were cultured in vitro and treated with 0,40,80 and 160 μg/mL EGCG, respectively. At the proposed time of treatment the morphological changes were detected by hoechst 33258 staining. The apoptosis rate was detected by flow cytometry. The expression of apoptosis-related factors B lymphocytoma-2 gene(bcl-2), BCL2-Associated X protein(Bax),caspase-3 and p53 were detected by quantitative RT-PCR and Western blotting.RESULTS: Hoechst 33258 staining showed that the ARPE-19 cells with the increase of EGCG drug concentration, the number of apoptotic cells gradually increased and the apoptotic bodies were observed. Flow cytometry showed that the apoptosis rate increased gradually with the increase of EGCG drug concentration. The apoptosis rates at 40, 80 and 160 μg/mL were 4.95%±0.071%, 11.75%±0.075% and 21.25%±0.919% respectively, which was significantly different compared with the control group(2.8%±1.556%)(P<0.01), presented with a drug concentration-dependent. The results of quantitative PCR and Western blotting showed that EGCG could significantly up-regulate the expression of apoptosis-promoting factors Bax, caspase-3 and the mRNA and protein expression of p53, and down-regulate the apoptosis-inhibiting factor bcl-2, all of these showed concentration-dependent effects.CONCLUSION:EGCG can obviously induce the apoptosis of ARPE-19 cells. The mechanism is related with the inhibition of bcl-2 and increase the expression of Bax, caspase-3 and p53.

Protective effects of naringin phospholipid complex on oxidative injury in ARPE-19 cells associated with activation of Nrf2 pathway / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the protective effects of naringin(Nar)phospholipid complex(NPC)on oxidative injury in retinal pigment epithelium cells(ARPE-19 cells)induced by tert-butyl hydroperoxide(t-BHP)and elucidate the underlying mechanism.<p>METHODS:The NPC was prepared by solvent method. Experimental cells are divided into seven groups: control group \〖cultured with dimethylsulfoxide(DMSO)\〗, model group(intervention with 200μmol/L t-BHP), nuclear factor erythroid 2-related factor 2(Nrf2)-siRNA group(cell transfection for Nrf2 gene), naringin group(add 200μmol/L t-BHP after pretreatment with 200μmol/L naringin medium), NPC group(add 200μmol/L t-BHP after pretreatment with 200μmol/L NPC medium), Nrf2-siRNA+ naringin group(after 200μmol/L naringin pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP)and Nrf2-siRNA+ NPC group(after 200μmol/L NPC pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP). The intracellular levels of superoxide dismutase(SOD), malondialdehyde(MDA)and total antioxidant capacity(T-AOC)were detected, intracellular level of reactive oxygen species(ROS)was detected by DCFH-DA staining method. The mRNA and protein expressions of HO-1, NQO-1, GCL and Nrf2 were detected by real-time PCR and western blot, respectively. <p>RESULTS:NPC more significantly increased the levels of SOD and T-AOC, reduced the contents of ROS and MDA than naringin in t-BHP-treated ARPE-19 cells. After naringin and NPC pre-protected ARPE-19 cells, the relative expression and protein expression of Nrf2, HO-1, NQO-1 and GCL mRNA were higher than those of the model group and Nrf2-siRNA group. There were statistically significant differences in the relative expression of 4 genes and the expression levels of 4 proteins in the naringin group and the NPC group, the Nrf2-siRNA+naringin group and the Nrf2-siRNA+NPC group. The expression of Nrf2, HO-1 and NQO-1 protein in the Nrf2-siRNA+naringin group was not significantly different than that in the Nrf2-siRNA group. Compared with the Nrf2-siRNA group, the expression of 4 proteins in the Nrf2-siRNA+NPC group was statistically significant, and the effect of NPC was significantly stronger than that of naringin.<p>CONCLUSION: After naringin forms a phospholipid complex, it can significantly increase the antioxidant capacity in cells and reduce the oxidation level. It up-regulates the expression of Nrf2 and its downstream antioxidant enzymes and phase Ⅱ detoxification enzymes by activating the Nrf2/ARE antioxidative stress pathway to better protect ARPE-19 cells from oxidative damage.

Dipenyleneiodonium Induces Growth Inhibition of Toxoplasma gondii through ROS Induction in ARPE-19 Cells

Based on the reactive oxygen species (ROS) regulatory properties of diphenyleneiodonium (DPI), we investigated the effects of DPI on host-infected T. gondii proliferation and determined specific concentration that inhibit the intracellular parasite growth but without severe toxic effect on human retinal pigment epithelial (ARPE-19) cells. As a result, it is observed that host superoxide, mitochondria superoxide and H2O2 levels can be increased by DPI, significantly, followed by suppression of T. gondii infection and proliferation. The involvement of ROS in anti-parasitic effect of DPI was confirmed by finding that DPI effect on T. gondii can be reversed by ROS scavengers, N-acetyl-L-cysteine and ascorbic acid. These results suggest that, in ARPE-19 cell, DPI can enhance host ROS generation to prevent T. gondii growth. Our study showed DPI is capable of suppressing T. gondii growth in host cells while minimizing the un-favorite side-effect to host cell. These results imply that DPI as a promising candidate material for novel drug development that can ameliorate toxoplasmosis based on ROS regulation.

Effect of blue light and white light on proliferation of ARPE-19 cells detected by CCK-8 / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effects of blue light and white light on the proliferation of human retinal pigment epithelial cell line(ARPE-19)at different times, and lay a foundation for further detecting the changes of related factors during photodamage and further studying the signal transduction mechanism during light damage. <p>METHODS:Well-grown ARPE-19 cells were collected for experimentation. The standard curve of CCK-8 was made to determine the proper cell density of ARPE-19 cells and the reaction time of CCK-8 reagent. The cells were divided into dark group, blue light group and white light group, which were irradiated for 3, 6 and 9h respectively. After 12h of light-repellent treatment, CCK-8 method was used to examine the effects of different light sources on the proliferation rate of ARPE-19 cells at different times. <p>RESULTS: The number of cells per well was selected by CCK-8 standard curve to be 20 000, and the corresponding absorbance value was measured after 4h of the action of CCK-8 solution. The CCK-8 test results showed that the cell proliferation rates of the three groups were significantly different(<i>P</i><0.01). The cell proliferation rate of the blue light group was significantly different(<i>P</i><0.001)at 3, 6 and 9h, and the cell proliferation rate decreased gradually with the extension of the illumination time. The cell proliferation rate of the white light group was significantly different(<i>P</i><0.05)at 3, 6 and 9h; there was a statistically significant difference in the rate of cell proliferation at 3h and 6h in white light(<i>P</i><0.05), however, there was no significant difference in the rate of cell proliferation at 9h illumination compared with 3h and 6h illumination respectively(<i>P</i>=0.253, 0.120). The proliferation rate of cells under white light for 3-6h showed a downward trend, while that of cells under light for 6-9h showed an upward trend. At the same illumination time, the proliferation rate of the cells in the blue and white groups was lower than that in the dark group, and the cell proliferation rate in the blue group was lower than that in the white group. The difference was statistically significant(<i>P</i><0.05). <p>CONCLUSION: The proliferation of ARPE-19 cells was inhibited by light irradiation. The proliferation rate of cells in blue light group was significantly lower than that of white light group. With the increase of light time, the cell proliferation rate decreased.

Modulated Gene Expression of Toxoplasma gondii Infected Retinal Pigment Epithelial Cell Line (ARPE-19) via PI3K/Akt or mTOR Signal Pathway

Due to the critical location and physiological activities of the retinal pigment epithelial (RPE) cell, it is constantly subjected to contact with various infectious agents and inflammatory mediators. However, little is known about the signaling events in RPE involved in Toxoplasma gondii infection and development. The aim of the study is to screen the host mRNA transcriptional change of 3 inflammation-related gene categories, PI3K/Akt pathway regulatory components, blood vessel development factors and ROS regulators, to prove that PI3K/Akt or mTOR signaling pathway play an essential role in regulating the selected inflammation-related genes. The selected genes include PH domain and leucine- rich-repeat protein phosphatases (PHLPP), casein kinase2 (CK2), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), glutamate-cysteine ligase (GCL), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase (NQO1). Using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), we found that T. gondii up-regulates PHLPP2, CK2β, VEGF, GCL, GST, and NQO1 gene expression levels, but down-regulates PHLPP1 and PEDF mRNA transcription levels. PI3K inhibition and mTOR inhibition by specific inhibitors showed that most of these host gene expression patterns were due to activation of PI3K/Akt or mTOR pathways with some exceptional cases. Taken together, our results reveal a new molecular mechanism of these gene expression change dependent on PI3K/Akt or mTOR pathways and highlight more systematical insight of how an intracellular T. gondii can manipulate host genes to avoid host defense.

Protective effect of Bu Shen Yang Xue Ming Mu Formula on oxidative injury in ARPE-19 cells / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the effect of Bu Shen Yang Xue Ming Mu (BSYXMM) Formula on hydroquinone-induced oxidative stress injury in ARPE-19 cells.METHODS: The oxidative injury model of ARPE-19 cell was induced by exposure to various concentrations of hydroquinone (HQ) to determine the optimal concentration.Intestinal absorption solutions of BSYXMM Formula were prepared.Effect of intestinal absorption solutions of BSYXMM Formula on the cell viability was detected by CCK-8 assay,and the percentage of apoptotic cells was measured by TUNEL assay.The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in ARPE-19 cells were detected by means of chemical colorimetry.RESULTS: It was found that ARPE-19 cell viability significantly decreased when the concentration of HQ was higher than 90μmol/L.Compared with the model group,1% and 2% intestinal absorption solutions in the pre-treatment group could significantly alleviate HQ-induced injury (P<0.01) and 0.5% and 5% intestinal absorption solutions in the pre-treatment group could alleviate the injury in certain degree(P<0.05).While in the treatment group 1% and 2% intestinal absorption solutions could alleviate the injury to some extent (P<0.05).TUNEL results showed that the apoptosis rate decreased significantly in the pre-treatment group (P<0.01)and to some extent in the treatment group (P<0.05)compared with the model group.It was shown that both levels of SOD and GSH-Px in pre-treatment group and treatment group were markedly higher than that of model group(P<0.05),and pre-treatment group had more significant effect (P<0.01,P<0.05).CONCLUSION: BSYXMM Formula could protect against HQ-induced oxidative stress injury in ARPE-19 cells,which may be related with the increasing of antioxidant enzyme in the cells.

Suppressors for Human Epidermal Growth Factor Receptor 2/4 (HER2/4): A New Family of Anti-Toxoplasmic Agents in ARPE-19 Cells

The effects of tyrosine kinase inhibitors (TKIs) were evaluated on growth inhibition of intracellular Toxoplasma gondii in host ARPE-19 cells. The number of tachyzoites per parasitophorous vacuolar membrane (PVM) was counted after treatment with TKIs. T. gondii protein expression was assessed by western blot. Immunofluorescence assay was performed using Programmed Cell Death 4 (PDCD4) and T. gondii GRA3 antibodies. The TKIs were divided into 3 groups; non-epidermal growth factor receptor (non-EGFR), anti-human EGFR 2 (anti-HER2), and anti-HER2/4 TKIs, respectively. Group I TKIs (nintedanib, AZD9291, and sunitinib) were unable to inhibit proliferation without destroying host cells. Group II TKIs (lapatinib, gefitinib, erlotinib, and AG1478) inhibited proliferation up to 98% equivalent to control pyrimethamine (5 μM) at 20 μM and higher, without affecting host cells. Group III TKIs (neratinib, dacomitinib, afatinib, and pelitinib) inhibited proliferation up to 98% equivalent to pyrimethamine at 1–5 μM, but host cells were destroyed at 10–20 μM. In Group I, TgHSP90 and SAG1 inhibitions were weak, and GRA3 expression was moderately inhibited. In Group II, TgHSP90 and SAG1 expressions seemed to be slightly enhanced, while GRA3 showed none to mild inhibition; however, AG1478 inhibited all proteins moderately. Protein expression was blocked in Group III, comparable to pyrimethamine. PDCD4 and GRA3 were well localized inside the nuclei in Group I, mildly disrupted in Group II, and were completely disrupted in Group III. This study suggests the possibility of a vital T. gondii TK having potential HER2/4 properties, thus anti-HER2/4 TKIs may inhibit intracellular parasite proliferation with minimal adverse effects on host cells.

The pilot study on rapamycin retarding the differentiation of RPE cells in vitro / 中华实验眼科杂志

Background Retinal pigment epithelial (RPE) cell transplantation is a novel approach to the treatment of hereditary retinal diseases, however, human-derived RPE cell line occurs de-differentiation during in vitro cell culture.Studies showed that early abnormal activation of Akt/mammalian target of rapamycin (mTOR) signal pathway is the primary cause of RPE cell line de-differentiation, therefore, the inhibition of mTOR pathway will be helpful for the retard of de-differentiation of RPE cells.Objective This study aimed to investigate whether rapamycin can suppress the activation of mTOR pathway and promote differentiation of ARPE19 cells.Methods ARPE-19 cells were incubated in 12-well plate and divided into control group and rapamycin-treated group.DMSO or rapamycin with the final concentration of 400 nmol/L was added in the medium of the control group and the rapamycin-treated group, respectively.The cells of each group were collected 24 hours and 48 hours after cultured.The expression of zonula occludens-1 (ZO-1) in the cells was examined by immunofluorescence.The relative expression levels of RPE cell specific genes and proteins were assayed by using real-time quantitative PCR and Western blot, respectively.The detected results were compared between the two groups.Results ZO-1 was expressed in both group,but the fluorescence intensity was evidently enhanced in the rapamycin-treated group.The relative expression levels of RPE65, MERKT and LRAT mRNA in the cells increased by 25.97％ , 29.71％ and 13.00％ in the rapamycin treated group compared with the control group 24 hours after cultured (P=0.04,0.04,0.04) , and the expression levels of RPE65, LRAT, rLBP1, BEST1 , keratin18 and MERKT mRNA elevated by 174.00％ , 88.00％ , 56.18％ ,193.81％ ,10.83％ and 35.02％ in the rapamycin-treated group in comparison with the control group 48 hours after cultured (P =0.00,0.04,0.01,0.04,0.04,0.03).In addition, the expressions of p-mTOR, p-P70S6 and p-S6 protein were weaker in the rapamyein-treated group than those in the control group both 24 hours and 48 hours after cultured.Twenty four hours after cultured,the expression level of ZO-1 protein raised by 40％ in the rapamycin-treated group compared with the control group (P =0.01);while 48 hours after cultured,the expression levels of ZO-1 ,MERKT, catenin and LRAT proteins elevated by 36.00％ ,57.37％, 13.68％ and 41.07％ in the rapamycintreated group in comparison with the control group (P=0.01,0.00,0.04,0.04).Conclusions Rapamycin can suppress the activation of mTOR signaling pathway and up-regulate the expressions of RPE specific genes in ARPE19 cells.Inhibition of mTOR pathway might be an effective way for culturing RPE cells in vitro.

Effects of the prescription of reinforcing kidney, nourishing blood, improving eyesight on ARPE - 19 cells induced by acrolein / 国际眼科杂志(Guoji Yanke Zazhi)

?AlM: To explore the effects of the prescription of reinforcing kidney, nourishing blood, improving eyesight on the oxidative stress model of ARPE-19 cells induced by acrolein. ?METHODS:SD rats serum containing the prescription of reinforcing kidney, nourishing blood, improving eyesight and the content of distilled water in serum were prepared. The effects of the prescription and distilled water in serum at different concentration ( 2. 5%, 5%, 10%, 20% and 40%) on cell vitality was observed by cell counting kit ( CCK-8 ) assay. the logarithmic phase of ARPE-19 cells were pretreated by different concentrations (1. 25%, 2. 5%and 5%) of the prescription serum and distilled water in serum for 24h. Then it was treated with 75μmol/L acrolein for 24h. Cell vitality was observed by CCK-8 assay. The change of cell nucleus was detected by DAPl staining . ?RESULTS: 2. 5% and 5% serum had no effect on cell viability (P>0. 05), while 10%, 20%, 40% serum could inhibit cell viability (P ?CONCLUSlON: The prescription of reinforcing kidney, nourishing blood, improving eyesight has the protective effect on ARPE-19 cell damage induced by acrolein.

2-ethylpyridine, a cigarette smoke component, causes mitochondrial damage in human retinal pigment epithelial cells in vitro.

Purpose: Our goal was to identify the cellular and molecular effects of 2-ethylpyridine (2-EP, a component of cigarette smoke) on human retinal pigment epithelial cells (ARPE-19) in vitro. Materials and Methods: ARPE-19 cells were exposed to varying concentrations of 2-EP. Cell viability (CV) was measured by a trypan blue dye exclusion assay. Caspase-3/7 and caspase-9 activities were measured by fl uorochrome assays. The production of reactive oxygen/nitrogen species (ROS/RNS) was detected with a 2’,7’-dichlorodihydrofl uorescein diacetate dye assay. The JC-1 assay was used to measure mitochondrial membrane potential (m). Mitochondrial redox potential was measured using a RedoxSensor Red kit and mitochondria were evaluated with Mitotracker dye. Results: Aft er 2-EP exposure, ARPE-19 cells showed signifi cantly decreased CV, increased caspase-3/7 and caspase-9 activities, elevated ROS/RNS levels, decreased m value and decreased redox fl uorescence when compared with control samples. Conclusions: These results show that 2-EP treatment induced cell death by caspase-dependent apoptosis associated with an oxidative stress and mitochondrial dysfunction. These data represent a possible mechanism by which smoking contributes to age-related macular degeneration and other retinal diseases and identify mitochondria as a target for future therapeutic interventions.

Synthesis of taurine-fluorescein conjugate and evaluation of its retina-targeted efficiency in vitro

In this work, retinal penetration of fluorescein was achieved in vitro by covalent attachment of taurine to fluorescein, yielding the F-Tau conjugate. Nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HRMS) were used to confirm the successful synthesis of F-Tau. The cellular uptake of F-Tau in adult retinal pigment epithelial cells (ARPE-19) and human retinal microvascular endothelial cells (hRMECs) was visualized via confocal scanning microscopy. The results indicated an improvement of solubility and a reduction of logP of F-Tau compared with fluorescein. As compared with fluorescein, F-Tau showed little toxicity, and was retained longer by cells in uptake experiments. F-Tau also displayed higher transepithelial permeabilities than fluorescein in ARPE-19 and hRMECs monolayer cells (P<0.05). These results showed that taurine may be a useful ligand for targeting small-molecule hydrophobic pharmaceuticals into the retina.

Effects of resveratrol on proliferation of ARPE-19 cells / 中国病理生理杂志

AIM:To investigate the effects of resveratrol ( Res) on the proliferation of ARPE-19 cells and to ex-plore the possible mechanisms.METHODS:After ARPE-19 cells were treated with Res at concentrations of 0, 50, 100, 150, 200 and 300 μmol/L for 24 h, 48 h and 72 h, the effects of Res on the proliferation of the cells were tested by CCK-8 assay.The ARPE-19 cells were treated with Res at concentrations of 0, 100, 150 and 200 μmol/L for 48 h.The effects of Res on the cell cycle and apoptosis were detected by flow cytometry with Annexin V-FITC/PI staining.The protein expression of proliferating cell nuclear antigen (PCNA) was detected by immunofluorescent assay.The mRNA expression of PCNA, P21 and P27 was determined by real-time PCR.RESULTS:The results of CCK-8 assay showed that Res inhibited the prolifera-tion of ARPE-19 cells in a time-and dose-dependent manner.The treatment with Res for 48 h resulted in an arrest of cell cycle at S phase without increasing cell apoptosis.Res inhibited the protein expression of PCNA in ARPE-19 cells.The re-sults of real-time PCR showed that Res increased the mRNA expression of P21 and P27, and decreased the mRNA expres-sion of PCNA.CONCLUSION: Res inhibits the proliferation of ARPE-19 cells and induces the cell cycle arrest at S phase.The mechanism may be related to up-regulation of P21 and P27, and down-regulation of PCNA.

Anti-apoptosis effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cells / 实用医学杂志

Objective To investigate the regulations of Bax , Bcl-2 in the protection of lipoic acid-niacin diad in acrolein-induced apoptosis in ARPE-19 cells. Methods The ARPE-19 cells were cultured in medium containing 10% fetal bovine serum , at 37 ℃ with 5% CO2. The ARPE-19 was transferred to 6-well plate after reaching to 70% confluence. After starvation for 24 h , the cells in 6-well plates were divided into three groups , including the blank control group , the acrolein treatment group with 50 μmol/L acrolein for 24 h , and the protection group with 100 μmol/L lipoic acid-niacin diad for 24 h and with the acrolein for another 24 h. The apoptotic cells were detected by flow cytometry assay , and expressions of Bcl-2 , Bax protein were detected by Western Blot assay. Results The percentages of normal healthy cells were 94.8%, 60.98%, and 91.34% in the blank control group , 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group , respectively. The ratios of Bax/Bcl-2 protein expression were 0.293 9, 1.389 2, and 0.555 8 in the blank control group, 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group, respectively. Conclusion The protective effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cell through promoting Bcl-2 expression and inhibiting Bax expression.

Role of Nitric Oxide in the Proliferative and Migratory Effect of Triamcinolone in RPE Cells

PURPOSE: To investigate the role of nitric oxide (NO) on the proliferative and migratory effects of triamcinolone acetonide (TA) in retinal pigment epithelial cells. METHODS: After exposure to 10 nM, 1 micrometer, or 100 micrometer TA for four days, with or without co-exposure of antioxidant N-acetylcyteine, the proliferation and nitrite production of ARPE19 cells were assessed with MTT and Griess assays, respectively. Additionally, a cell migration assay was performed. RESULTS: Cellular survival increased after exposure to TA at low concentration but decreased at high concentration. TA decreased the production of NO and cellular migration significantly, and these effects were abolished by N-acetylcysteine. CONCLUSIONS: TA showed a biphasic response on the proliferation and decreased cellular migration in ARPE19 cells, which may be mediated by nitric oxide.

Antioxidant activity of naringenin on various oxidants induced damages in ARPE-19 cells and HUVEC / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the antioxidant activity of naringenin in human retinal pigment epithelium (ARPE-19) cells andhuman umbilical vein endothelial cells (HUVEC).·METHODS: MTT assay was used to measure theviability and proliferation of ARPE-19 cells and HUVEC.·RESULTS: Three and 10mg/L naringenin significantlyincreased the proliferation of ARPE-19 cells by 10.8% and11.4%, respectively. Ten mg/L naringenin increasedhypoxia-, 0.3mmol/L NAN3-, and 200μmol/L H2O2- induced damage of ARPE-19 cells by 55.2%, 69.2%, and50.3%, respectively. One mg/L naringenin increased theviability of 50μmol/L t-BHP-, and 30mg/L NalO3-treatedARPE-19 cells by 20.2% and 30.4%, respectively. Thirtymg/L naringenin also increased the proliferation of50μmol/L t-BHP-treated ARPE-19 cells by 32.2%, and1mg/L naringenin increased the proliferation of 30, 100and 300 mg/L NalO3-treated ARPE-19 cells by 30.3%,10.3% and 18.5%, respectively. The reduction of HUVECwas 23.9%, 70.4% and 77.9% in the 3, 10 and 30mg/Lnaringenin-treated groups, respectively. Furthermore, 1and 3mg/L naringenin increased hypoxia-induced damagein HUVEC by 10.7% and 13.1%, and 300mg/L NalO3- induced damage in HUVEC by 41.2% and 37.7%. Threemg/L naringenin increased 200 and 400μmol/L H2O2-in-jured HUVEC by 20.1% and 21.5%, respectively.·CONCLUSION: Naringenin increases the proliferation ofARPE-19 cells and inhibites the growth of HUVEC, and haspotent antioxidant activity in ARPE-19 cells and HUVEC.