| Objective:Through the successful established rat model of diabetic retinopathy,discussion the effects of Yougui Wan which based on " reinforcing yang from yin" for VEGF-VEGFR pro-survival signaling pathways in diabetic retinopathy rat. Methods:80 SPF SD rats were selected, 10 rats set up as normal group; the remaining 70 rats were given streptozotocin(60mg/kg)raperitoneal injection, to establish the model of diabetic rats. After 4 weeks, treated diabetic rat VEGF 0.05 ug real injection caused by rat model of proliferative diabetic retinopathy period; After 4 weeks, and each row fundus fluorescein angiography in rats, according to angiography results will eventually molded filter 40 rats were randomly divided into model group, YouguiWan high, medium and low dose group, n = 10; the corresponding doses were given orally, the normal group and model group were given the dose and other doses of distilled water. Three months later, each group of rats underwent eye retina retinal tissue histopathology was observed; RT-PCR was used to detect the retina VEGFmRNA, VEGF2 mRNA expression; Western-blot assay retinal VEGF, VEGFR-2activity protein Level. result:1, The establishment of animal models: after STZ injection 72 h,1 week,2 weeks,3 weeks,all the rats blood sugar stable tendency in ≧16.7 mmol/l,successfully established the diabetic model; diabetic rats vitreous body cavity injection of VEGF fundus fluorescence angiography revealed after 4 weeks,the rat model group were observed in the backgroud fluorescence enhancement,blood vessels, circuity expansion,such as fluorescent leakage is simillar to human proliferation period clinical pathological changes of diabetic retinopathy.2. Pathological examination in retinal tissue of rats showed: the layers of the rules of normal rats retina, no abnormal change of retinal morphology; the rats in model group showed different degrees of edema, loose structure, and granular cell swelling, derangement disorder, and dilatation and congestion of the capillaries, scattered in the outer plexiform layer the pathological neovascularization; YouguiWan low dose group and model group of rat retinal changes are similar; YouguiWan medium and high dose group of retinal layers are arranged opposite rule neat, edema, both inside and outside cells arranged in more regular particle layer, outer plexiform layer even visible expansion, congestion of the capillaries, no neovascularization.3.By RT-PCR method to detect rat retina VEGFmRNA, VEGFR-2mRNA expression Results: Compared with the normal group, model group and treatment group VEGFmRNA, VEGFR-2mRNA expression enhanced, the difference was statistically significant(P< 0.01); compared with the model group, YouguiWan medium and high dose group VEGFmRNA, VEGFR-2mRNA are lower, the difference was statistically significant(P<0.05), no significant changes in Youguiwan low dose group(P ﹥ 0.05); compared with the YouguiWan low dose group, YouguiWan medium and high dose group VEGFmRNA, VEGFR-2mRNA expression decreased, the difference was statistically significant(P <0.05),compare YouguiWan medium with high dose groups, VEGFmRNA, VEGFR-2mRNA expression have no significant changes(P> 0.05).4. By Western-blot method to detect rat retina VEGF, VEGFR-2 protein expression Results: Compared with the normal group, model group and treatment group VEGF, VEGFR-2 VEGF, VEGFR-2 protein expression enhanced, the difference was statistically significant(P < 0.01); compared with the model group, YouguiWan medium and high dose group VEGF, VEGFR-2 protein are lower, the difference was statistically significant(P <0.05), no significant changes in Youguiwan low dose group(P ﹥ 0.05); compared with the YouguiWan low dose group, YouguiWan medium and high dose group VEGF, VEGFR-2 protein expression decreased, the difference was statistically significant(P <0.05),compare YouguiWan medium with high dose groups, VEGF, VEGFR-2 protein expression have no significant changes(P> 0.05).conclusion:1 By intraperitoneal injection of STZ, the intravitreal injection of VEGF, established rat model consistent with proliferative diabetic retinopathy and human performance in the clinic, is to establish a viable method of proliferative diabetic retinopathy animal model.2 YouguiWan diabetic retinopathy retinal layers of cell proliferation and microvascular damage in rats have improved to some extent, and reduce VEGFmRNA, VEGFR-2mRNA expression, reduced VEGF, VEGFR-2 protein levels to downstream signaling pathways in the regulation of VEGF expression of protein activity.3 YouguiWan of DR development of PDR Qi yin and yang to the virtual machine to disease " reinforcing yang from yin", kidney yang, may promote survival signaling pathway by regulating VEGF-VEGFR, delaying the process of development of diabetic retinopathy. |
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