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The Antiangiogenic Effect of Vasostatin on Diabetic Retinopathy (DR) in Mice

Author: FengWeiYan
Tutor: GeYingBin
School: Nanjing Medical University
Course: Physiology
Keywords: vasostatin Diabetic retinopathy Human umbilical vein endothelial cells Angiogenesis Apoptosis
CLC: R774.1
Type: Master's thesis
Year: 2009
Downloads: 28
Quote: 0
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Abstract


Diabetic retinopathy (Diabetic Retinopathy, DR) is the most common microvascular complications of diabetes, duration of more than 10 years the incidence of diabetic patients with DR up to 80%, has become the world's most important one of the blinding eye disease. DR pathological mechanisms in retinal vascular abnormalities newborn blood - retinal barrier breakdown and retinal detachment characterized. Therefore, the anti-angiogenesis drugs and changes in diet are considered to be an effective method for the prevention and treatment of diabetic retinopathy. Vasostatin calreticulin (calreticulin) N 1-180 amino acid sequence of the fragment of the segment. As an endogenous angiogenesis inhibitor, vasostatin inhibit tumor angiogenesis and tumor proliferation efficacy has been confirmed, there are certain similarities in view of the pathogenesis of diabetic retinopathy and tumor, vasostatin may diabetic retinopathy angiogenesis inhibited. To test this hypothesis, we designed the following two-part experiment. 1. Streptozotocin (STZ)-induced diabetic mouse model detection vasostatin whether inhibition of diabetic mice retinal angiogenesis. 8-week-old C57BL/6J mice were divided into diabetic group and the control group, diabetic group in accordance with the dose of 100mg/kg body weight for 5 consecutive days built intraperitoneal injection of STZ diabetic mice, the control group in the same way given solvent. After 16 weeks, STZ-induced diabetic mice successfully modeling is selected, the control group using the same amount of PBS vasostatin 100ng/ml soluble medical sodium hyaluronate gel daily eye drops administered three consecutive weeks . Flash ERG (FERG), record the Ops-wave detection mouse retina vascular function while using FITC-labeled Griffonia Simplicifolia lectin (Isolectin B4) staining of retinal microvascular morphological changes. The FERG detection ERG in diabetic mice, the the Ops wave amplitude is significantly lower than the control group, significantly prolonged peak latencies. Isolectin B4 staining showed that diabetic mice retinal vascular proliferation, growth in total vessel length, vessel branch increased significantly compared with the control group. After to give vasostatin treatment, these phenomena were significantly improved. Using in vitro cultured human umbilical vein endothelial cells (HUVEC) study the vasostatin antiangiogenic mechanism of action. HUVEC cells with different concentrations vasostatin processing hypoxia 24 hours after CCK-8, and Hoechst 33342 staining cell survival as well as the number of apoptotic cells. Western-blot detection of cell-associated apoptosis signaling pathways protein expression. The results showed that hypoxia, the number of apoptosis in HUVEC cells was positively correlated with vasostatin concentration, dose-dependent. Western-blot results showed that the inhibition of Akt phosphorylation in HUVEC cells, vasostatin raised Bad phosphorylation activate casapse-3 activity to induce apoptosis. The study suggests that the topical application vasostatin reduce diabetic retinopathy angiogenesis and improve retinal vascular dysfunction caused by diabetic retinopathy. Antiangiogenic role of Vasostatin may be related to the inhibition of growth factor-dependent apoptosis pathway Akt/Bad/caspase-3. In short, the results suggest that, vasostatin become one of the potential drug effective in treating diabetic retinopathy, but its mechanism of action and safety needs further study.

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CLC: > Medicine, health > Ophthalmology > Retina and optic nerve diseases > Retinal disease
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