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Biomedical subjects

Hua He

Publications and source records attributed to Hua He.

At least 19 recordsLinked to original sources
PubMed2026

Developing Highly Effective Nanoparticle mRNA Therapeutic for Pediatric Acute Respiratory Distress Syndrome.

Sepsis-induced pediatric acute lung injury (ALI) and pediatric acute respiratory distress syndrome (PARDS) are life-threatening conditions with high mortality rates and no current cure. Most ALI/ARDS studies focus on adults, albeit the pediatric population has unique challenges often underrepresented. ALI/PARDS severely impacts pulmonary endothelial cells (ECs), causing endothelial dysfunction and vascular leakage. FOXF1 is a transcription factor critical for lung repair after injury, representing a viable target for ALI/PARDS. This study developed and tested a novel nanoparticle system for precise delivery of FOXF1 mRNA into lung ECs to reduce endothelial damage and improve lung function in mouse model of PARDS. Systemic inflammatory response was induced in neonatal mice after intraperitoneal administration of lipopolysaccharide (LPS). Specifically designed nanoparticles (NP) were used to intravenously deliver stabilized FOXF1 mRNA (FOXF1 NP) after LPS injury to restore FOXF1 expression in injured lung endothelial cells. FOXF1 NP selectively targeted pulmonary ECs without affecting other cell types or organs. FOXF1 NP treatment reduced vascular leakage, enhanced endothelial barrier function, and improved survival of neonatal mice after injury. FOXF1 NP decreased EC apoptosis by restoring the expression of BCL2, an anti-apoptotic FOXF1 target gene. Nanoparticle-based rescue of lung ECs has promise for future treatments of human ALI/PARDS.

endothelial cells↗
PubMed2006

POT1b protects telomeres from end-to-end chromosomal fusions and aberrant homologous recombination.

POT1 (protection of telomere 1) is a highly conserved single-stranded telomeric binding protein that is essential for telomere end protection. Here, we report the cloning and characterization of a second member of the mouse POT family. POT1b binds telomeric DNA via conserved DNA binding oligonucleotide/oligosaccharide (OB) folds. Compared to POT1a, POT1b OB-folds possess less sequence specificity for telomeres. In contrast to POT1a, truncated POT1b possessing only the OB-folds can efficiently localize to telomeres in vivo. Overexpression of a mutant Pot1b allele that cannot bind telomeric DNA initiated a DNA damage response at telomeres that led to p53-dependent senescence. Furthermore, a reduction of the 3' G-rich overhang, increased chromosomal fusions and elevated homologous recombination (HR) were observed at telomeres. shRNA mediated depletion of endogenous Pot1b in Pot1a deficient cells resulted in increased chromosomal aberrations. Our results indicate that POT1b plays important protective functions at telomeres and that proper maintenance of chromosomal stability requires both POT proteins.

Amino Acid Sequence↗
PubMed2006

Correlation between the single-site CpG methylation and expression silencing of the XAF1 gene in human gastric and colon cancers.

BACKGROUND & AIMS: X-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (XAF1) antagonizes the anti-caspase activity of XIAP. XAF1 messenger RNA is present in normal tissues but undetectable in various cancers and thus poses a potential tumor suppressor gene. The aim of this study was to examine the novel pattern of methylation of XAF1 in gastric and colon cancers and locate the important CpG sites for transcriptional regulation and tumor progression. METHODS: XAF1 expression was detected by reverse-transcription polymerase chain reaction (PCR) and Western blot analysis. Four different fragments around the transcription start site of XAF1 were cloned and examined putative promoter activities by luciferase reporter assay. Each CpG site in fragment F291 was mutated by site-directed mutagenesis technique, and the change of promoter activity of this fragment was detected by luciferase reporter assay. Methylation status of XAF1 was determined by methylation-specific PCR (MSP) and bisulfite DNA sequencing PCR analysis. RESULTS: Down-regulation of XAF1 in association with hypermethylation was detected in 3 of 4 human gastric cancer cell lines and 6 of 8 colon cancer cell lines. Of the 4 promoter fragments, F291 showed the highest promoter activity, which could be down-regulated obviously by the mutation of particular CpG sites. Moreover, aberrant hypermethylation of these important CpG sites was strongly associated with the development of gastric and colon cancers. CONCLUSIONS: A cluster of methylated CpG sites instead of CpG islands located in the promoter area resulted in gene silencing of XAF1, and CpGs at -2nd, -1st, and +3rd positions are functionally more important in its transcriptional regulation.

Adaptor Proteins, Signal Transducing↗
PubMed2006

Ginsenoside-Rd from panax notoginseng blocks Ca2+ influx through receptor- and store-operated Ca2+ channels in vascular smooth muscle cells.

Previously, it was found that total saponins from panax notoginseng inhibited Ca2+ influx coupling to activation of alpha1-adrenoceptor. This study was designed to investigate the effects of ginsenoside-Rd from total saponins of panax notoginseng on receptor-operated (ROCC) and store-operated (SOCC) Ca2+ channels in vascular smooth muscle cells using fura-2 fluorescence, whole cell patch clamp ion channel recording, radio-ligand-receptor binding, 45Ca2+ radio-trace and organ bath techniques. It was found that ginsenoside-Rd reduced phenylephrine-induced contractile responses and Ca2+ influx in normal media without significant effect on these responses in Ca2+ -free media. Ginsenoside-Rd also decreased phenylephrine- and thapsigargin-induced inward Ca2+ currents, and attenuated thapsigargin- and 1-oleoy-2-acetyl-sn-glycerol (OAG)-induced cation entries that are coupled to ROCC and SOCC respectively. Ginsenoside-Rd failed to inhibit KCl-induced contraction of rat aortal rings and Ca2+ influx, and did not alter voltage-dependent inward Ca2+ current (VDCC) which was blocked by nifedipine. Also, ginsenoside-Rd did not change binding site and affinity of [3H]-prazosin for alpha1-adrenoceptor in the vascular plasma membrane. These results suggest that ginsenoside-Rd, as an inhibitor, remarkably inhibits Ca2+ entry through ROCC and SOCC without effects on VDCC and Ca2+ release in vascular smooth muscle cells.

Animals↗
PubMed2006

Pot1 deficiency initiates DNA damage checkpoint activation and aberrant homologous recombination at telomeres.

The terminal t-loop structure adopted by mammalian telomeres is thought to prevent telomeres from being recognized as double-stranded DNA breaks by sequestering the 3' single-stranded G-rich overhang from exposure to the DNA damage machinery. The POT1 (protection of telomeres) protein binds the single-stranded overhang and is required for both chromosomal end protection and telomere length regulation. The mouse genome contains two POT1 orthologs, Pot1a and Pot1b. Here we show that conditional deletion of Pot1a elicits a DNA damage response at telomeres, resulting in p53-dependent replicative senescence. Pot1a-deficient cells exhibit overall telomere length and 3' overhang elongation as well as aberrant homologous recombination (HR) at telomeres, manifested as increased telomere sister chromatid exchanges and formation of telomere circles. Telomeric HR following Pot1a loss requires NBS1. Pot1a deletion also results in chromosomal instability. Our results suggest that POT1a is crucial for the maintenance of both telomere integrity and overall genomic stability.

Animals↗
PubMed2006

Inhibition of Akt/PKB by a COX-2 inhibitor induces apoptosis in gastric cancer cells.

BACKGROUND/AIM: Inhibition of cyclooxygenase-2 has been proposed to be a potential mechanism for the chemoprevention of gastrointestinal tumors by nonsteroidal anti-inflammatory drugs. This study investigates the mechanisms by which the cyclooxygenase-2 inhibitor SC236 induces apoptosis of gastric cancer cell lines and its downstream signaling pathway. METHODS: Two gastric cancer cell lines, AGS and MKN28, were treated with SC236 and assessed for cell growth and apoptosis. The involvement of mitogen-activated protein kinase and Akt kinase/protein kinase B (Akt/PKB) pathways and their downstream signalings were studied in the AGS cell line. RESULTS: SC236 treatment induced apoptosis in gastric cancer cells and caused activation of p38 and stress-activated protein kinase/jun kinase, but down-regulated Akt/PKB. The specific p38 inhibitor SB203580 and the dominant-negative stress-activated protein kinase/jun kinase both failed, while the constitutively active form of Akt/PKB was able to block SC236-induced apoptosis. SC236-induced apoptosis was coupled with release of cytochrome c and activation of caspases. CONCLUSION: One of the pathways involved in SC-236-induced apoptosis in gastric cancer cells is through downregulation of Akt and then release of cytochrome c.

Acridine Orange↗
PubMed2006

Inhibition of human telomerase reverse transcriptase by nonsteroidal antiinflammatory drugs in colon carcinoma.

BACKGROUND: Telomerase activation, which is observed in most human cancers, plays an important role in carcinogenesis. Human telomerase reverse transcriptase (hTERT) is a subunit of telomerase that is essential for telomerase activity. The aim of the study was to investigate whether nonsteroidal antiinflammatory drugs (NSAIDs) inhibit telomerase activity and hTERT. METHODS: Four colon carcinoma cell lines, HT-29, COLO205, CRL-2134, and SW1116, were used in the experiments. Polymerase chain reaction-based telomeric repeat amplification (TRAP) enzyme-linked immunosorbent assay (ELISA) was used to measure telomerase activity in the cells after treatment with aspirin, indomethacin, or SC-236 (a specific cyclooxygenase-2 [COX-2] inhibitor). Expression of hTERT mRNA and protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The dual luciferase reporter assay was performed to identify the potential cis-response elements to NSAIDs in the promoter region of hTERT. RESULTS: Aspirin, indomethacin, and SC-236 inhibited telomerase activity in HT-29, COLO205, and CRL-2134 cell lines, but not in the SW1116 cell line. NSAIDs inhibited hTERT mRNA and protein expression through suppression of hTERT transcriptional activity. The hTERT promoter fragment -145 to -330 basepairs (bp) upstream of the ATG starting site was sufficient to respond to the NSAID-induced inhibitory effect and the inhibition was COX-2-independent. CONCLUSION: NSAIDs inhibit telomerase activity at hTERT transcriptional, mRNA, and protein levels in colon carcinoma cells. The hTERT promoter fragment -145 to -330 bp may be the cis-response element to NSAIDs.

Adenocarcinoma↗
PubMed2006

Fosinopril and carvedilol reverse hypertrophy and change the levels of protein kinase C epsilon and components of its signaling complex.

OBJECTIVE: To demonstrate the alterations of Protein Kinase C epsilon (PKC epsilon) and components of its signaling complexes after treatment with fosinopril and carvedilol and analyze potential molecular mechanisms of the two drugs for cardiac hypertrophy and heart failure. METHODS: Pressure-overload cardiac hypertrophy (POH) was developed in 8-week-old male Sprague Dawley rats by abdominal aortic banding. The rats were divided into three groups at the age of 20 weeks: POH without failure group, reversed POH with drugs group, and POH with failure group on high diet. Western Blot analysis, co-immunoprecipitation and proteomic analysis were performed in ventricular tissues of rat hearts. RESULTS: Increased PKC epsilon was found during POH. PKC epsilon decreased during transition from POH to heart failure (HF). However, increased PKC epsilon inclined to recover to normal levels after treatment with both drugs. There were differential proteins in PKC epsilon complexes during the different stages of POH. The two significant PKC epsilon-binding proteins, MAD1 and Lyn A, were only present in PKC epsilon complex during reversing POH with drugs. CONCLUSION: Chronic administration of carvedilol and fosinopril could reverse the development of POH and delay the appearance of HF, partly by regulating PKC epsilon level and its signaling complex. MAD1 and Lyn A may be important proteins participating in the reversing process.

Adrenergic alpha-Antagonists↗
PubMed2006

All-trans retinoic acid induces XAF1 expression through an interferon regulatory factor-1 element in colon cancer.

BACKGROUND & AIMS: X-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (XAF1) is a novel tumor suppressor and interferon (IFN)-stimulated gene. All-trans retinoic acid (ATRA) exerts an antiproliferative effect on tumor cells through up-regulation of IFN regulatory factor 1 (IRF-1) and the downstream IFN-stimulated genes. The aim of this study was to determine the effect and mechanism of ATRA on XAF1 expression and the role of XAF1 in ATRA-induced growth inhibition in colon cancer. METHODS: Gene expression is detected by reverse-transcription polymerase chain reaction and immunoblotting. The transcription activity of XAF1 promoter is examined by luciferase reporter assay. The activity of IFN regulatory factor binding element (IRF-E) is assessed by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. Cell growth is evaluated by both in vitro and in vivo in nude mice xenografts. RESULTS: IFN-alfa stimulates XAF1 promoter activity in the colon cancer cells Lovo and SW1116 dose-dependently. An IRF-1 binding element (IRF-E-XAF1) is found in the -30 to -38 nucleotide region upstream of the ATG initiator codon of the XAF1 gene. Site-directed mutagenesis of IRF-E-XAF1 abrogates native and IFN-induced promoter activity and binding capacity. ATRA induces XAF1 expression both in vitro and in vivo through interaction with IRF-E-XAF1. Overexpression of XAF1 increases cell susceptibility to ATRA-induced growth suppression both in vitro and in vivo. Furthermore, the effect of ATRA on XAF1 expression is independent of the promoter methylation and the subcellular distribution of XIAP. CONCLUSIONS: XAF1 participates in ATRA-induced growth suppression through IRF-1-mediated transcriptional regulation.

Adaptor Proteins, Signal Transducing↗
PubMed2006

Determination of vertilmicin sulfate and its related substances by HPLC-ELSD and HPLC-MS2.

A new and simple high-performance liquid chromatography (HPLC)-evaporative light scattering detection (ELSD) method for the determination of vertilmicin sulfate and its related substances is developed. The column is an Agilent SB-C(18) (250 x 4.6 mm, 5 microm). The mobile phase is 0.05 mol/L trifluoroacetic acid-methanol (85:15). Good separation of vertilmicin from the main related substances is achieved. The standard curve is rectilinear in the range of 270-1350 microg/mL (r = 0.9998). The average recovery is 99.8%. The limit of detection is 10 microg/mL. The HPLC-mass spectrometry-mass spectrometry (MS(2)) method is used to characterize the structures of vertilmicin sulfate and its related substances. In positive mode, vertilmicin sulfate and its related substances are elucidated by use of electrospray ion trap MS in the multi-stage MS full scan mode. The possible structure of an unknown impurity in vertilmicin is deduced based on the HPLC-MS(2) data.

Aminoglycosides↗
PubMed2006

Interaction between Cl- channels and CRAC-related Ca2+ signaling during T lymphocyte activation and proliferation.

AIM: To test the hypothesis that Cl channel blockers affect T cell proliferation through Ca2+-release-activated Ca2+ (CRAC) signaling and examine the effects of the combination of a CRAC channel blocker and a Cl channel blocker on concanavalin A (ConA; 5 mg/mL)-induced Ca2+ signaling, gene expression and cellular proliferation in human peripheral T lymphocytes. METHODS: [3H]Thymidine incorporation, Fura-2 fluorescent probe, RNase protection assay, and reverse transcription-polymerase chain reaction were used. RESULTS: The Cl channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) inhibited ConA-induced Ca2+ influx, interleukin-2 mRNA expression and T lymphocyte proliferation in a concentration-dependent manner, and also enhanced the inhibitory effects of 1-[beta-[3-(4-methoxyphenyl)propoxyl]-4-methoxyphenethyl]-1H-imidazole (SK&F96365) on the above key events during T cell activation. A combination of DIDS (1 micromol/L) and SK&F96365 (1 micromol/L) significantly diminished ConA-induced ClC-3 mRNA expression by 64%, whereas DIDS(1 micromol/L) or SK&F96365 (1 micromol/L) alone decreased ConA-induced ClC-3 mRNA expression by only 16% and 9%, respectively. CONCLUSION: These results suggest that there is an interaction between CRAC-mediated Ca2+ signaling and DIDS-sensitive Cl channels during ConA-induced T cell activation and proliferation. Moreover, the DIDS-sensitive Cl channels may be related to the ClC-3 Cl channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2006

Signaling-transduction pathways required for ex vivo expansion of human limbal explants on intact amniotic membrane.

PURPOSE: Ex vivo expansion of limbal epithelial progenitor cells on amniotic membrane (AM) without 3T3 fibroblasts is a new surgical approach to treat limbal stem cell deficiency. Such expansion requires NGF-TrkA-mediated signaling, and this study was conducted to delineate the downstream signaling pathways. METHODS: The human corneolimbal ring was cut into explants and cultured on intact human AM. At day 0 or 10, low-molecular-weight inhibitors were added, whereas the control group received dimethyl sulfoxide (DMSO). The epithelial outgrowth rate was monitored for 17 days, and the epithelial cells were collected for Western blot analysis. RESULTS: In the control, most expansion of human limbal epithelial cells started from the limbus from days 5 to 7 and reached approximately 80% confluence at day 17. Compared with the control, the outgrowth was completely inhibited by 50 microM LY294002 or 50 microM SR13668 and was significantly suppressed by 10 microM U0126, but was not affected by 10 microM of either SB203580 or JNK inhibitor 1. The inhibition of outgrowth by LY294002, SR13668, and U0126 was reversible. Western blot analysis showed that phosphorylation of Akt and FKHRL1was abolished by LY294002 and SR13668, but downregulated by U0126, which also abolished phosphorylation of p44/42 mitogen-activated protein kinase (MAPK). The phosphorylation of p38 and JNK MAPK were downregulated or abolished during ex vivo expansion. CONCLUSIONS: Ex vivo expansion of human limbal epithelial progenitor cells on intact AM is mediated by the survival signaling pathway mediated by PI3K-Akt-FKHRL1 and by the mitogenic MAPK pathway mediated by p44/42 at the expense of p38 and JNK MAPK.

Amnion↗
PubMed2006

Preservation and expansion of the primate keratocyte phenotype by downregulating TGF-beta signaling in a low-calcium, serum-free medium.

PURPOSE: To demonstrate whether the original keratocyte phenotype is maintained with proliferative activity by suppressing TGF-beta signaling in rhesus monkey keratocytes expanded in a serum-free and low-[Ca2+] medium. METHODS: Rhesus monkey keratocytes were isolated from central corneal buttons by collagenase digestion for 16 hours, seeded on plastic in Dulbecco's modified Eagle's medium (DMEM) containing insulin-transferrin-sodium selenite (ITS) supplement (DMEM/ITS) or 10% fetal bovine serum (DMEM/10% FBS), or in a defined keratinocyte serum-free medium (KSFM). After confluence, cells in KSFM were continuously subcultured at a 1-to-3 split. Cellular proliferation was analyzed by immunostaining for Ki67 and the MTT assay. The cellular phenotype was determined by immunostaining for aldehyde dehydrogenase (ALDH), keratocan, and CD34 and by the expression of keratocan promoter-driven enhanced cyan fluorescent protein (ECFP). The stability of the keratocyte phenotype was examined by switching KSFM to DMEM/ITS and DMEM/10% FBS. TGF-beta signaling was monitored by measuring the promoter activity of TGF-beta1, -beta2, and -beta RII after transient adenoviral transfection, and cytolocalization of Smad2 and Smad4. RESULTS: In KSFM, monkey keratocytes proliferated while maintaining the expression of keratocan, CD34, and ALDH proteins and keratocan promoter-driven ECFP for at least 15 passages. The nuclear accumulation of Smad2 and Smad4 and the promoter activities of TGF-beta1 and -beta RII were significantly downregulated in KSFM compared with DMEM/10% FBS. In KSFM, an increase of [Ca2+] to 1.8 mM and addition of 10% FBS synergistically downregulated the keratocan promoter activity, facilitated Smad2 and Smad4 nuclear translocation, and upregulated TGF-beta1 and -beta RII promoter activities. CONCLUSIONS: The normal monkey keratocyte phenotype can be maintained in a low-calcium, serum-free medium by downregulating Smad-mediated TGF-beta signaling.

Adenoviridae↗
PubMed2006

Basement membrane dissolution and reassembly by limbal corneal epithelial cells expanded on amniotic membrane.

PURPOSE: To investigate basement membrane (BM) formation during ex vivo expansion of limbal corneal epithelial cells on intact amniotic membrane (iAM) and epithelially denuded (d)AM. METHODS: Human limbal explants were cultured on iAM and dAM. Expression of BM components, including laminin-5, type IV collagen, type VII collagen, perlecan, integrin alpha6, and epithelial cell differentiation markers such as p63, cytokeratin 3 (K3), and cytokeratin 12 (K12), were investigated by immunostaining. Levels of matrix metalloproteinase (MMP)-2 and MMP-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in the conditioned media were determined by ELISA and gelatin zymography. RESULTS: All four BM components were preserved in both iAM and dAM before culturing, but dissolved 1 week afterward when MMP-2 was increased. Epithelial outgrowth correlated with increased expression of MMP-2 and -9 for both cultures. Resynthesis of BM began with laminin-5 followed by other components. This process took place at 1 week on iAM but at 2 weeks on dAM after culturing. At 4 weeks, BM was more maturely deposited as a linear band from the explant toward the leading edge on iAM and temporally correlated with a sharp decline of MMP-9 levels. In contrast, such BM deposition began at the leading edge on dAM only when TIMP-1 levels were increased. Epithelial cell outgrowth on iAM expressed more p63 but less K3 and K12 than did that on dAM. CONCLUSIONS: After dissolution of original amniotic BM, new BM formed by ex vivo expanded human limbal corneal epithelial cells on iAM deposits much faster and is more mature, resulting in regeneration of a limbal epithelial phenotype. In contrast, BM deposition is delayed and remains immature on dAM, resembling wound healing by a corneal epithelial phenotype. Thus, BM resynthesis may be used as another objective readout for assessing the success of ex vivo expansion of limbal epithelial progenitor cells on AM.

Amnion↗
PubMed2006

[Study on the interaction of lomefloxacin terbium complex and bovine serum albumin by fluorescence spectra].

Using lomefloxacin-terbium as a fluorescent probe, the interaction of LFLX-Tb3+ and bovine serum albumin (BSA) was investigated by fluorescence spectra. The interaction of LFLX and BSA is quite strong. LFLX can affect the conformation of BSA to some degree. Meanwhile, the electrostatic binding of the BSA with Tb3+ in LFLX-Tb3+ complex could decease the nonradiative energy loss through O-H vibration of H2O molecule in Tb3+ complex. Under the optimum conditions, BSA can enhance the fluorescence intensity of the lomefloxacin-terbium complex at 545 nm, and the enhanced fluorescence intensity of Tb3+ is proportional to the concentration of BSA. Based on that, a new method is constructed for the determination of BSA. The linear ranges for BSA are 16.5-148.5 microg x mL(-1) with a detection limit of 68.8 ng x mL(-1) and relative standard deviations of 1.4%. This method is simple, practical and relatively free of interference from coexistent substances.

Animals↗
PubMed2006

[Fluorescence characteristic of terbium-ciprofloxacin complex and its application].

On binding to deoxyribonucleic acid, the complex of terbium-ciprofloxacin (Tb(3+)-CIP) increases its fluorescence quantum efficiency. Based on this, an easy, rapid and sensitive method for the determination of DNA was developed. Like ethidium bromide (EB), the complex can be intercalated into DNA bases, but it is non-poisonous. Determination can be made at pH 7.0, where the native structure of DNA is not disrupted. The maximum emission is at 545 nm with excitation at 325 nm. This method has good sensitivity (2.8 x 10(-9) mol x L(-1) of ctDNA), high selectivity and a wide linear range (4.3 x 10(-7)-3. 0 x 10(-5) mol x L(-1) of ctDNA). This complex was also employed for the clinical examination of DNA in whole blood of tumor patients. Ten tumor patients and 10 normal persons were enrolled in this study. It was showed that significant difference existed between control group and tumor group (p < 0.05).

Animals↗
PubMed2006

[Inhibitory effect of RGDS peptide on experimental choroidal neovascularization in a rat model].

OBJECTIVE: To investigate the efficacy and feasibility of RGDS (Arg-Gly-Asp-Ser) peptide (an alpha(nu)-integrin antagonist) in a rat model of laser-induced choroidal neovascularization (CNV). METHODS: Experimental CNV was induced by laser photocoagulation in Brown Norway rats (50 microm diameter, 0.05 second duration and 525 mw intensity). Phosphate buffered saline (PBS) or 100, 300 microg of RGDS peptide in PBS were being injected intravitreally after laser surgery for 7 days. On the 14th day after photocoagulation, CNV was observed by fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA). The area of CNV by high molecular weight FITC-dextran (MW 2 x 10(6)) for high resolution angiography in RPE-choroid-sclera flat mounts and the thickness microscopically on histologic sections were evaluated. New vessels were detected and quantified by an-antibody against factor VIII. Two eyes from each group were examined by transmission electron microscopy. RESULTS: In eyes with injections of 300 or 100 microg of RGDS peptide on the 14th day after laser photocoagulation, the development of CNV was significantly (P < 0.01) inhibited showing by RPE-choroid-sclera flat mounts. Histologically, the thickness of the CNV lesions was significantly (P < 0.01) reduced in eyes that received 300 or 100 microg of RGDS peptide injection. Immunoreactivity of factor VIII in CNV showed significant difference (P < 0.01) in eyes injected with RGDS peptide compared with control eyes. The reduction of the area and the thickness of CNV by RGDS peptide were in a dose-dependent manner. No evidence of toxicity was found in retina by transmission electron microscopy in every group. CONCLUSIONS: RGDS peptide effectively inhibits CNV progression in a rat model of laser-induced CNV, suggesting that this alpha(nu)-integrin antagonist may be beneficial in the treatment of CNV.

Animals↗