水蛭渗滤液对猴RF/6A细胞表达TNF-α的影响

Effect of leech extract on thrombin induced monkey RF/6A cell in expressing angiogenesis-promoting factor TNF-α

目的观察水蛭渗滤液对凝血酶诱导下猴脉络膜-视网膜(内皮)细胞(RF/6A)表达促血管生成因子TNF-α的影响。方法采用消化培养法,对猴RF/6A细胞进行体外传代培养,选择20μ/ml的凝血酶作为诱导浓度,分别作用于RF/6A细胞2、4、6、8、10、12及24 h,用ELISA法观察凝血酶对视网膜血管内皮细胞表达TNF-α的诱导情况。观察在有或无凝血酶诱导下水蛭渗滤液(64 mg/ml)对视网膜血管内皮细胞表达TNF-α的影响。结果 (1)凝血酶作用2 h,TNF-α的表达开始增加,作用4 h,TNF-α的表达达到最大,持续至10 h;4、6、8 h与空白组比较差异均有统计学意义(P<0.05);当凝血酶作用时间达12 h,TNF-α的表达开始下降,凝血酶组与空白组12 h和24 h比较,差异无统计学意义(P>0.05)。(2)水蛭渗滤液对有或无凝血酶诱导组都能下调TNF-α的表达(P<0.05)。结论水蛭渗滤液能下调凝血酶诱导下猴RF/6A细胞表达TNF-α。

OBJECTIVE To observe the affection of Leech extract to RF/6A cell by thrombin-induced and to investigate the mechanism of preventive affection of retinal choroidal neovascularization. METHODS RF/6A cell cultured in vitro by digestion culture, and induced by thrombin （20 μ/ml, 2,4,6,8,10,12,24 hour） to observe the ex- press of TNF-ot cytokine. Simultaneously, we observed the express of TNF-α by leech extract （64 mg/ml） to RF/6A cell, with or without thrombin-induced. RESULTS The leech extract could decline the TNF-α express of RF/6A cell, with or without thrombin-induced. CONCLUSIONS Leech extract could prevent the retinal vascular disease, from a certain extent, by blocking thrombin activation to reduce retinal angiogenesis factor TNF-α expression to achieve.