| Background:retinal vein occlusion (RVO) is the second common retinal vascular disease after Diabetic retinopathy, which is also one of the main blind eye disease in China.The etiology and pathogenesis of RVO is extremely complex. It’s difficult to treat it. However It can get good curative effect by using the methods of Liangxue Zhixue and Huoxue Tongluo under the theory of "Xue Zheng Lun", but the mechanism of these two methods,when and how they work have not yet clear.Objective:To investigate the mechanism of Liangxue Zhixue and Huoxue Tongluo of RVO rabbit model about the retina microcirculation and blood coagulation related factors.Methods:Rabbits for randomized controlled trials, establishing by photochem method.78New Zealand white rabbits randomly divided into blank group, model group, Zhixue group (group A), Huoxue group (group B), Zhixue and Huoxue group (group C), five groups. Blank group6rabbits, without building models, only give physiological saline; The rest of the four groups, each one include18rabbits, raise different herbs after building models.Take photos of eye ground and do FFA in order to observe and record leakage area, retina fluorescent cycle time and the recanalization of block veins, when30minutes later after building models and the1st,3rd,7th,14th,28th day after taking herbs.Test the level of antithrombin Ⅲ, plasminogen, TXB2and6-Keto-PGF1a.Results:1The performance of rabbit eye ground and FFA:1.1the different period of rabbit eye ground:On the1st day, the performance of A, B and C is similar to model group; The3th～7th day three groups have different degrees of bleeding absorption, group A is more apparent; After the14th day, bleeding is absorbed mostly of A, B and C, no obvious edema, and the condition of blood vessels is better than model group, group B and C are better than A; After the28th day, the performance of A, B and C is similar to model group.1.2the different period of rabbit FFA:On the1st day, the performance of A, B and C is similar to model group; The3rd～7th day the area of blocked flourescene and Limitations high fluorescence of A, B and C are smaller than model group, group A is more apparent, few of rabbit from B and C get vein recirculated; After the14th day, blocked flourescene disappear mostly of A, B and C, some rabbits have hemangioma and ramusanastomoticus, parts of them get vein recirculated, no significant difference among A, B and C; After the28th day, the performance of A, B and C is similar to model group.1.3FFA vascular recanalization:None of the groups recirculate during30minutes to the1st day after building models; vascular recanalization appeared in group A, B and C, none of model group recanalizated, on the3rd day; On the7th day it appeared in model group, and blood become more unobstructed in group A, B and C; On the14th day vascular recanalization appeared in half of model group and all of chinese medicine groups; On the28th day, blood is unobstructed in all groups.2Observation of rabbit eyes through optical microscope in different period:After the3rd day, the situation of thrombus and NFLof group A, B and C is better than model group, B’s thrombus is less than A and C, A’s NFL is better than B and C, B bleed more than A and C, but similar to model group; On the14th day, the arrangement of ganglion cells in group A, B and C is better than model group, A and B is better than C.3Observation of rabbit eyes through electron microscope in different period:After the3rd day, the structure of nucleus and cytoplasm of group A, B and C is better than model group, A is the best; On the14th day, group A, B and C is better than model group, A and B is better than C. On the28th day, group A, B and C is better than model group.4the level of ATⅢ in different period:There’s significant difference between model group and group A (P<0.05), from30minutes to1day after building models; On the3rd day, it’s between model group and group B (P<0.01), between group A and group B, group C (P<0.01); On the7th day, it’s between model group and group C (P<0.01), between group B and group C (P<0.01); From the14th to28th day no significant difference between each two groups.5the level of PLG in different period:there’s no significant difference between each two groups, from30minutes to1day after building models; On the1st day, there’s significant difference between group B and model group, group A (P<0.05); On the3rd day, it’s between model group and group A, group B, between each two chinese medicine groups (P<0.01); On the7th day, it’s between group A and model group, group C (P<0.01); From the14th to28thday it’s between group B and group A, group C (P<0.05).6the level of TXB2in different period:On the3rd day, there’s significant difference between model group and group B (P<0.01), between group C and group A, B (P<0.05); On the14th day, it’s between group B and other three groups (P<0.01); On the28th day, no significant difference between each two groups. It showed that from the14th day to28th day every group can increases the level of TXB2.7the level of TXB2/6-Keto-PGF1a in different period:On the3rd day, there’s significant difference between model group and group C (P<0.01); On the14th day, it’s between group B and group A, C (P<0.05); On the28th day, it’s between model group and three chinese medicine groups (P<0.01).Conclusion:1It showed that the changes of retina tissues of photochemical models is just like the clinical manifestation of RVO from FFA and pathological results.2Devided RVO into3stages according to the time, within7d is the early one, around14d is the middle one,28d is the late one. Liangxue Zhixue and Huoxue Tongluo method can promote the absorption of retina edema and bleeding, and enhance vessel situation. Zhixue method is better for early stage (within7d), act on absorbing edema and bleeding of retina, while Huoxue method is better for the late-mid stage (after14d), act on enhancing vessel situation. The effect is similar to Huoxue method when using these two methods together.3Liangxue Zhixue and Huoxue Tongluo method can Protect the3kinds of retina nerve cells and Slow down or reduce pathological changes of cells, with varying degrees. Huoxue method Promote thrombolytic in the early stage (within7d), Zhixue method reduce Ischemic Changes of nerve fiber in the early stage (within7d); Zhixue and Huoxue method Protect gangliocyte in the middle stage (around14d); Zhixue and Huoxue method reduce the progressive changes of nerve cells and RPE in the late stage (around28d).The effect is worse than using the two method individually, when using them together, in all stages.4The pathological changes of retinal ultrastructure is consistent with tissue’s, after intervention of Chinese medicine. In the early stage (3d), it happens on all levels of the nerve cells.5The changes of blood coagulation factor and retina capillaries of model group: Anticoagulant factor is lower than normal level, appearing blood coagulation state,in the early and middle stages (within14d); It returned to normal gradually, after middle stage (after14d); Retina capillaries contract in the early stage (3d), then gradually ease. It begins to relax after middle stage (after14d).6Compared with model group, Liangxue Zhixue and Huoxue Tongluo methods have fuction on blood coagulation factor and microvascular smooth muscle, but the mechanism is different. Using them together does not mean add each of their mechanism simply.7Liangxue Zhixue method regulate and control blood coagulation factor and vascularsmoothmuscle in two ways. In the aspect of blood coagulation mechanism, Liangxue Zhixue method promote coagulation to stop bleeding, in the early stage(within3d); After7days it has anticoagulation to reduce thrombosis. In the aspect of adjusting vascular smooth muscle:Liangxue Zhixue method mainly effect on relaxing blood vessel, in the early and middle stages, which may improve the hypoxia, and promote microcirculation reconstruction. After the middle stage appears contraction vascular trend8Huoxue Tongluo method regulate and control blood coagulation factor in one way. It has anticoagulation to reduce thrombosis, in the early and middle stages; After middle stage this function is reduced. In the aspect of adjusting vascular smooth muscle:in the early, it has the trend of vasodilation, but the effect is not obvious.In the late-mid stage it can relax vessel.9Using two methods together can mainly effect on blood coagulation factor to promote coagulation, in the early and middle stages. In the other stages neither anticoagulation nor coagulation is obvious. In the aspect of adjusting vascular smooth muscle:The function of vasodilation is stronger than Liangxue Zhixue and Huoxue Tongluo method, in the early stage; After middle stage, the function of relaxing vessel is obvious, but similar to Huoxue Tongluo method.10From the mechanism, it showed the function of blood coagulation in the early stage of Liangxue Zhixue and the early and middle stage of two methods; it showed the function of anticoagulation after the middle stage of Liangxue Zhixue and Huoxue Tongluo method. In the aspect of adjusting micrangium:The three methods have similar effect on micrangium, in two ways. Relaxing vessel in the early stage, while constricting it in the late stage. Liangxue Zhixue method can Relax vessel for a longer time. Huoxue Tongluo method and two methods constrict vessel earlier.11It showed that Liangxue Zhixue and Huoxue Tongluo method effect on blood coagulation factor and microcirculation in two ways, It is different from western medicine, so it plays various function according to difrernt disease, stage and pathology. This suggests that we should treat patients on the basis of differentiation. |
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