| Objective:We used Yiqiwenyangtongmai Decoction with different doses on the model of Anterior Ischemic Optic Neuropathy (AION) rats, to observe the changes of flash visual evoked potential (F-VEP) and the optic disc and optic nerve in morphology, so as to explore the optic nerve protective effect and mechanism about Yiqiwenyangtongmai Decoction in ischemia condition.Method:60 male Sprague-Dawley rats were divided into 5 groups randomly:blank group, model group, traditional Chinese medicine high dose group, medium dose group and low dose group, with 12 rats in each group. Except the blank group, the rest rats of the 4 groups were injected in Rose Bengal 2.5mmol/(ml · kg) by caudal vein,2/3 of the area above the optic disc was irradiated with the wave length of 532 nm, power of 75mW, and spot diameter of 500μm green laser for 18 seconds immediately. To determine whether there were successful models or not, 1day after the building of models, we used the fundus camera and fluorescence fundus angiography (FFA) to observe the optic disc and its surrounding change of rat; And we used F-VEP to observe the change of the incubation period (ms) and amplitude(u v) of P2 wave respectively. After the models were made successfully, the rats of model group were given 2ml of distilled water everyday. High, medium and low dose group were given 2ml of the water decoction of traditional Chinese medicine, made of different concentration; The rats of blank group were given normal breeding. All groups were given intervention for 21 days. We used F-VEP to observe the change of the incubation period (ms) and amplitude(u v) of P2 wave respectively after intervention. After F-VEP check finished, the eyeballs of rats were removed to research by histological observation.Results:1. Fundus Photograph:There was no obvious unusual conditionon the optic disc and vessel around it in normal rat; 1 day after the building of models, there was obvious edema upper part on optic disc and the border of optic disc was unclear.2. FFArThere was no obvious fluorescence leakage on the optic disc and vessel around it in normal rat; 1 day after the building of mode Is, there was hypofluo rescence upper part on optic disc in early pha se and hyperfluo rescence in metaphase and later phase.3.Flash Visual Evoked Potential (F-VEP):The incubation periods and amplitudes of P2 wave were different between the blank group and the other four groups (P<0.01), and except the blank group, there were no difference in four groups each other before intervention(P>0.05). After intervention of 21 days, comparing with model group, there were significantly different in incubation periods and amplitudes of P2 wave of the blank group and high dose group (P<0.01), except other two groups(P>0.05). And the comparison in each group before and after intervention of 21 days, there was a statistical significance only in high dose group(P<0.01).4. Hematoxylin-Eosin (HE) staining:Intervention of 21 days, in the blank group, the structure of organization around the optic disc was complete. The morphology of optic nerve cell was normal, optic nerve fibers were linear and neatly; In the model group, the structure of organization was destructed seriously around the optic disc and the retina was been moved. Optic nerve cells were proliferated significantly, and the arrangement was very disordered, vacuoles of degeneration clearly, optic nerve fibers were atrophic; In the high dose group, the structure of organization around optic disc was complete. The morphology of optic nerve cell was normal and optic nerve fibers were linear, but nuclear column was crowded; The performance of medium dose group and low dose group were similar to the model group.5. Immunohistochemistrystaining:Intervention of 21 days, in the blank group, SMI312 was expressed obviously in axons; In the model group, expression of SMI312 was weak, which could be found only in the surrounding of optic nerve axons, expression of SMI312 was missing on optic nerve central part; In the high dose group, SMI312 was also expressed visibly in axons; The performance of medium dose group and low dose group were similar to the model group.6. TdT-mediated Nick End Label ing (TUNEL):Intervention of 21 days, in the blank group, optic nerve tissue did not see apoptosis cell; In the model group, TUNEL positive cells were expressed obviously on optic nerve. Compared with the model group, TUNEL positive cells of blank group decreased significantly (P<0.01); In the high dose group, TUNEL positive cells were very few on optic nerve tissue, there was significant difference between high dose group and model group (P<0.01); In the medium dose group and low dose group, TUNEL positive cells were expressed clearly on optic nerve, and compared with model group, there was no big difference(P>0.05).Conclusions:1. Yiqiwenyangtongmai Decoction had a significantly protective effect on the structure and function in AION rats, but it needed to reach a certain concentration.2. The reason may be related with promoting the repair of optic nerve axon fibers and save optic nerve cells in time, which were not completely apoptosis. |
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