Diabetes is a disease caused by high blood sugar levels that occur when the pancreas does not secrete enough insulin or the body is unable to process insulin properly.
Patients with diabetes can develop eye problems such as cataracts and glaucoma, but the disease’s effect on the retina is the main threat to vision. The effect of diabetic retinopathy on vision varies widely, depending on the stage of the disease. Diabetic patients require regular eye examinations so related eye problems can be detected and treated early.
The diagnosis of diabetic retinopathy is made following a detailed examination of the retina. Diabetic retinopathy is treated depending on the stage of the disease and the specific problem that requires attention. If the presence of abnormal blood vessels is detected, a small application of laser treatment (Focal Laser Treatment) or a wider area of laser treatment (panretinal photocoagulation-PRP) may be the treatment of choice. With PRP, the ophthalmologist applies laser to reduce the amount of oxygen-deprived retinal tissue outside of the patient’s central vision. Vitrectomy surgery may be needed for diabetic patients who suffer a vitreous hemorrhage (bleeding in the gel-like substance that fills the center of the eye).
Diabetics can also greatly reduce the possibilities of eye complications by scheduling regular examinations with an ophthalmologist. Rand Eye Institute’s Vitreo-Retinal Department offers comprehensive medical and surgical management for vitreo-retinal diseases with the most advanced technology. Diabetic retinopathy is a common diabetic eye disease caused by changes in retinal ganglion cells (RGCs).
Effects of superoxide (DHE), MDA and 8-OHdG levels and MnSOD activity in the retinal tissue.
The effect of astaxanthin on H2O2-induced apoptosis in the transformed rat retinal ganglion cell line RGC-5.
AcknowledgementsThis work was financially supported by The National Natural Science Foundation of China (No.
Even patients with well-controlled diabetes can develop diabetic changes in the retina after years. Diabetic patients need to be under the close observation of an internist or an endocrinologist as well as by an ophthalmologist. In severe cases, patients with diabetic retinopathy may be referred to a retinal specialist. The ophthalmologist may rely on several special tests to monitor the progression of the disease and to make decisions for the appropriate treatment.
While this may affect the peripheral vision, PRP can prevent the continued growth of the fragile vessels and seal the leaking ones, thereby preserving vision.
During a vitrectomy, the surgeon carefully removes blood and vitreous from the eye and replaces it with a clear natural-like solution. Rand is Chairman of the Florida Board of the Diabetes Research Institute (DRI) Foundation and serves as the Vice-Chair of the national board. DRI scientists are making tremendous progress toward a cure, however, until we reach our ultimate goal, it is important for patients to practice excellent diabetes management to maintain optimal health.
Close collaboration with the other Rand Eye Institute departments assures that patients will receive timely and personalized retinal as well as comprehensive ophthalmology services. It is an ocular manifestation of systemic disease, which affects up to 80% of all patients who have had diabetes for 10 years or more.
IntroductionDiabetic retinopathy is well-known as the result of microvascular retinal changes [1]. Astaxanthin Attenuates H2O2-Induced Apoptosis in the Transformed Rat Retinal Ganglion Cell Line RGC-5More and more research pinpoints oxidative stress as a root cause of diabetic retinopathy [23,24]. DiscussionAs nature’s most powerful antioxidant, astaxanthin has been documented to show a wide range of benefits in human clinical studies on several serious health concerns [25]. 81271726) and The Research and innovation project of the Shanghai Municipal Education Commission (No. In this phase, the arteries in the retina become weakened and leak, forming small, dot-like hemorrhages. New fragile vessels grow as the circulatory system attempts to maintain adequate oxygen levels within the retina. These tests include: fluorescein angiography, retinal photography, macular OCT testing and ultrasound imaging of the eye. The goal of the treatment is to slow or stop the progression of the deterioration of vision.
At the same time, the surgeon may also gently remove strands of vitreous attached to the retina that could lead to retinal detachment or tears.
Retinal detachment requires surgical treatment to reattach the retina to the back of the eye.
The sole mission of the DRIF is to support the Diabetes Research Institute at the University of Miami Leonard M.
Hyperglycemia-induced intramural pericyte death and thickening of the basement membrane leads to incompetence of the vascular walls.
Dysfunction induced by the decreased population of cells is regarded as an important factor in the pathogenesis of various metabolic diseases [18,19]. To estimate the anti-oxidative effect of astaxanthin, superoxide anion (DHE), MDA and 8-OHdG levels and MnSOD activity in the retina tissues were measured.


To investigate the effect of astaxanthin on the apoptosis of RGC-5 cells, cell viability was determined in cells using MTT assays. Due to its molecular structure, astaxanthin is one of the few antioxidants that can move throughout the entire body and provide protection to all of our cells.
12ZZ101) and The Pudong New Area Science and Technology Development Fund innovative capital projects (No. If the disease progresses, leaking vessels can lead to swelling (edema) in the retina and decreased vision. Miller School of Medicine, a recognized world leader in diabetes research and the best hope for a cure for diabetes. Initial symptoms should not be overlooked and an immediate consultation with a vitreo-retinal specialist can make all the difference. Astaxanthin is a carotenoid with powerful antioxidant properties that exists naturally in various plants, algae and seafood. These damages change the formation of the blood-retinal barrier and also make the retinal blood vessels become more permeable [2]. To determine the effects of astaxanthin on RGCs, the apoptosis was determined using tunel assays. Apoptotic cells were marked with green fluorescence, the nuclei of cells are stained by blue fluorescence (DAPI) and RGCs are stained by red fluorescence (Neu, neuron).
As an initial indicator of ROS, we used DHE staining, which is a probe for O2•? and produces red fluorescence as a result of the complex between ethidium and DNA as described by previous study [22]. Astaxanthin significant inhibited the decrease of RGC-5 cell viability induced by H2O2 in a dose-dependent manner (Figure 4A). Data are expressed as % of Annexin V-FITC- positive and PI-negative cells (early stage of apoptosis).
PKJ2011-Y320) and The Shanghai Jiaotong University Biomedical Engineering (PolyU) Cross Research Fund Project (No.
However, recent studies have shown that the retina has a high content of polyunsaturated fatty acids and has the highest oxygen uptake and glucose oxidation relative to any other tissue. Retinal electroretinogram (ERG) was recorded from 8 mice per group, at 12–14 weeks of age in response to a series of light flashes at increasing intensity. The detection of DNA damage by tunel staining indicated that astaxanthin decreased the apoptosis of RGCs in retina (Figure 2). To quantitate changes in O2•? levels, we measured total DHE fluorescence in retinal homogenates. In addition, astaxanthin attenuated hydrogen peroxide(H2O2)-induced apoptosis in the transformed rat retinal ganglion cell line RGC-5.
Treatment with astaxanthin for 8 weeks reduced the expression of apoptosis gene proteins in the retina as determined by Western blot (C). Therefore, astaxanthin may be developed as an antioxidant drug to treat diabetic retinopathy.
And many diabetes-induced metabolic abnormalities were implicated in its development, and appeared to be influenced by elevated oxidative stress; however, the exact mechanism of its development remained elusive [4].
Furthermore, pretreatment with astaxanthin prevented H2O2-induced apoptosis in a dose-dependent manner. Before the oxygen consumption test (E), RGC-5 cells were pre-treated in low oxygen conditions for 2 h in suspension culture with astaxanthin and H2O2.
In addition, administration of astaxanthin might be a novel approach for the prevention of diabetes nephropathy [29]. Retina and capillary cells experience increased oxidative damage in the diabetic milieu, and the antioxidant defense mechanism is impaired [5].
In hyperglycemia condition, the over-oxidation of lipid or glucose leads to increase the level of reactive oxygen species and induce oxidative stress in tissues. Taken together, it suggested that astaxanthin had a strong anti-apoptotic effect by inhibition of H2O2-induced oxidative stress in RGC-5 cells. Our results provided a new insight into potential beneficial effects of antioxidants in the treatment of retinopathy in diabetic patients. The possible sources of oxidative stress in diabetes might include auto-oxidation of glucose, shifts in redox balances, decreased tissue concentrations of low molecular weight antioxidants such as reduced glutathione (GSH) and vitamin E, and impaired activities of antioxidant defense enzymes such as MnSOD (manganese superoxide dismutase) and catalase [6].
In addition, oxidative stress inactivated VEGF survival signaling in retinal endothelial cells via PI 3-kinase tyrosine nitration [7].
Moreover, astaxanthin attenuated H2O2-induced apoptosis in the transformed rat retinal ganglion cell line RGC-5. Therefore, oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species and a biological system’s ability to readily detoxify the reactive intermediates or to repair the resulting damage.
Reactive oxygen species (ROS) generated by high glucose are considered as a causal link between elevated glucose and the other metabolic abnormalities important in the development of diabetic complications [8].Apoptosis is a form of programmed cell death, which enabled a cell to direct its own destruction. It is well-known that the development of diabetes mellitus is caused by chronic multi-factors such as Gluco-lipotoxicity, inflammatory mediators and oxidative stress [30,31]. This form of cell death appeared to be crucial for mammalian development and subsequent tissue homeostasis.
Many diabetes-induced metabolic abnormalities are implicated in its development, and appear to be influenced by elevated oxidative stress; the exact mechanism of its development remains elusive.
In addition, oxidative stress induced caspase-independent apoptosis of retinal ganglion cells in vitro [9]. In our studies, astaxanthin could decrease the level of oxidative stress that caused dysfunction, apoptosis and ATP depletion.


Ceramide is the key mediator of oxidative stress-induced apoptosis in retinal photoreceptor cells [10]. Apoptosis had been evidenced directly and the loss of functional cells was observed in diabetic patients’ retina. Thus, the induced apoptosis, such as in retinal ganglion cells or photoreceptor cells, played an important role in diabetic retinopathy.Astaxanthin is a naturally occurring carotenoid with strong antioxidant properties both in vitro and in vivo [11]. In addition, we think astaxanthin, as an antioxidant, shows its radical scavenging ability by decomposition of H2O2. It has been reported that astaxanthin restored the enzymatic antioxidant profile in salivary gland of alloxan-induced diabetic rats [12], and protected retinal cells against oxidative stress in vitro and in mice in vivo [13].
However, astaxanthin significantly improved oscillatory potentials (OPs) in vivo in Figure 1.
The oscillatory potentials (OPs) in electroretinogram (ERG) have clinical values in measuring retinal functions of the early stage of diabetic retinopathy [32]. The National Eye Institute data is very alarming; it suggests that about half of the people with diabetes in the United States have at least some form of retinopathy, and about 700,000 have some serious retinal disease. Diabetic retinopathy is affecting approximately 65,000 people in the United States alone causing 12,000 to 24,000 new cases of blindness each year [3].
Moreover, oxidative stress played the key role on diabetic retinopathy, which directly caused cell dysfunction or apoptosis in retinal ganglion cells (RGCs) [33,34] and activated VEGF survival signaling in retinal endothelial cells via PI 3-kinase tyrosine nitration [35]. Thus, oxidative stress and antioxidant defenses may be as a target for the treatment of Diabetic Eye Disease.As shown in Figure 3, astaxanthin reversed the change of oxidative stress markers in the retinal tissue, including superoxide anion (DHE), MDA and 8-OHdG and MnSOD. Astaxanthin treatment resulted in a decline of blood fat level but the differences were not statistically significant among the experimental groups (Figure 1F).
To evaluate the effect of astaxanthin on glucose metabolism, random-blood glucose test were conducted after treatment with astaxanthin for 2 h. As shown in Figure 1G, astaxanthin could not improve the impaired glucose tolerance and increase glucose uptake in vivo.
All tissues were rapidly removed, placed in liquid nitrogen and stored at ?80 °C for subsequent preparation of pathological sections. Animal studies were carried out according to the Guidelines for Animal Experiments, as outlined by the Committee for Animal Experiments of National Center. Electroretinogram AnalysisThe b wave amplitude was measured from the a-wave negative peak to the positive peak of the b-wave. If the peak of the b-wave coincided with the oscillatory potentials, the point halfway between the trough and peak of the highest OP was used as the b-wave amplitude.
Oscillatory potentials were isolated from the averaged recording traces using a 75–300 Hz digital filter in the Espion E2 system. Amplitude of the oscillatory potentials was measured from bottom of the trough preceding each OP peak, to the top of that peak. The response threshold was defined as the minimum stimulus intensity that elicits a response greater than the criterion amplitude. Blood Glucose TestRandom tests were performed at every week after the onset of Astaxanthin dosing. Approximately 5 ?L of whole blood was drawn from the tip of the tail vein, and glucose was measured with the Optium Xceed™ Diabetes. After protein content determination using a DC Protein Assay kit (Bio-Rad Laboratories, Hercules, CA, USA), western blot was performed as described previously [42].
Detection of the Level of Superoxide Anion (O2•?), MDA, 8-OHdG and MnSODWe used the probe dihydroethidium (DHE; Molecular Probes, Eugene, OR, USA) to detect intracellular O2•?. The average concentration of 8-OHdG per microgram of DNA for each group was calculated for each sample. Controls without added DNA and appropriate blanks were also incorporated into experiments.The enzyme activity of MnSOD was measured in 5–10 ?g of retinal protein using a kit. The method utilizes tetrazolium salt to quantifying O2•? generated by xanthine oxidase and hypoxanthine.
MTT AssayCell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Briefly, the cells were seeded in 96-well dishes at 1 ? 104 to 2 ? 104 cells per well, and pretreated with or without astaxanthin for 2 h. The medium was then removed, and 150 ?L dimethyl sulfoxide (Sigma) were added to solubilize the MTT formazan. Staining with Hoechst allows for the discrimination of apoptotic cells based on nuclear morphology and evaluation of membrane integrity.
Oxygen ConsumptionTo evaluate the ability of cellular oxygen consumption (VO2) during drug treatment, the Micro Respirometry System (Strathkelvin, Mitocell S200) was used to measure oxygen content of culture media. Before the assay, primary hepatocytes just isolated from mice were pre-treated in low oxygen conditions for 2 h in suspension culture with drugs. Therefore, astaxanthin may be developed as a new drug to treat Diabetic Eye Disease from oxidative stress.



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