Clinical Review Of Liangxuehuayu Formula On The Treatment Of Exudative Age Related Macular Degeneration And The Study Of Its Mechanism

[Objective]1. To sum up and analyze the long-term therapeutic effect of modified Liangxuehuayu formula on the treatment of exudative age related macular degeneration.2. To study the effect and the mechanism of Mingjing granule on the experimental CNV.[Methods]1. Patients from Jan.2005to April2010treated by Dr. Tang youzhi with Liangxuehuayu formula for exudative age-related macular degeneration were reviewed and assessed. Bringing in and ruling out criterias were set up. Visual acuity, retinal manifestation and OCT were measured at first visit,1month after treatment (AT),3months AT,6months AT and12months AT.2. The method of subretinal injection of Matrigel was used to build up the model of experimental CNV. Fundus examination, FFA, choroidal flat mount and pathological changes were measured to choose the best dose and injecting technique of Matrigel.3.40grey rabbits were randomly divided into control group, model group, high dose of Mingjingkeli group, middle dose of Mingjingkeli group and small dose of Mingjingkeli group. The latter4groups were subretinal injected with Matrigel.28days later, fundus examination, FFA, tissue slides were measured; CD14and CD206possitive cells were measured by immunohistochemistry.4.70C57BL/6mice were used to build up the laser induced CNV model, frost tissue slides and immunofluorescence double labers techniques were used to measure the CNV growth、VEGF expression、macrophages and its phenotype around laser spots, the effect of Mingjingkili on the above aspects was also valued.[Results]1. Visual acuity of patients who received the treatment of Liangxuehuayu formula was improved after1month therapy(p<0.05), the rate of visual acuity increased at least two lines after3months treatment was16.13%. Retinal hemorrhage and exudation was alleviated or cured among84.34%patients, average retinalthickness was also extenuated after3months treatment (p<0.05).2. In the FFA examination, the fluorescence leakage was happened in Matrigel40μl and20μl groups at7d, which was enhanced at14d and28d; the fluorescence leakage was happened in Matrigel10μl group at14d and28d, which was never found in Matrigel5μl group.In the fundus examination, most of retina were attached back at7d, a few of those were detached. A few of retina had proliferative detachment in Matrigel40μl group. Cataract and vitreous muddy were found in Matrigel40μl group. In the tissue pathological examination, small vessels and/or immature fibrous tissues were found in Matrigel40μl and20μl groups. The retinal injure was most severely in Matrigel40μl group.3. In the FFA examination, all of the large, middle and small doses of Mingjingkeli could decrease the area of fluorescence leakage (p<0.05). In the tissue pathological examination, Mingjingkeli had the protective effect on the retina which was injected with Matrigel, the middle dose is the best, the large and small doses were the second. Based on the immunohistochemistry, there had many CD14+cells(monocyte)(p<0.05), less CD206+cells(type Ⅱ macrophage) in model group, while there had the same CD14+cells and much more CD206+cells in Mingjingkeli group(p<0.05).4. In the laser induced experimental CNV model, CD105+endothelial cells could be seen in7d, increased in14d and28d. VEGF was seen in neovascular masses in7d, was most obvious in14d and decreased in28d. F4/80+cells (mature macrophages) in neovascular masses were increased with time, most of those were CD32+cells (type Ⅰ macrophages), there were some CD206+cells in28d in model group. Mingjingkeli could decrease the growth of neovascularization, decrease the expression of VEGF, most of macrophages were CD206+cells. [Conclusion]1. Liangxuehuayu formula could improve the visual acuity by diminishing the retinal hemorrhage, exudation and edema, the visual acuity could be kept stable within12months.2. The method of subretinal injection of Matrigel could create CNV successfully,20μl is the best dose.3. All three doses of Mingjingkeli could inhibit CNV growth, the middle dose is the best one.4. Macrophages could be seen in the formation of CNV in both models of subretinal injection of Matrigel and laser induced CNV, most of which are type Ⅰ macrophages; Mingjingkeli could decrease the expression of VEGF, derivate the production of type Ⅱ macrophages.