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目的观察活血解毒方对糖尿病大鼠视网膜中Hippo信号通路的影响。方法选用健康SD大鼠24只,禁食12 h,一次性腹腔注射链脲佐菌素(65 mg/kg),诱导糖尿病模型。将造模20周后的大鼠随机分为模型组和活血解毒方组。另选取10只大鼠作为正常对照组,给予等体积蒸馏水灌胃。每组每天干预1次,共12周。分别采用免疫组织化学方法、蛋白质印迹(Western blot)方法检测各组视网膜中磷酸化哺乳动物不育系20样激酶(P-MST)、大肿瘤抑制基因1(Lats1)、磷酸化Yes-相关蛋白(P-YAP)、转录相关因子(TAZ)、TEA结构域转录因子(TEAD)蛋白表达水平的变化。结果免疫组织化学方法的结果显示:与正常组比较,模型组大鼠视网膜中P-MST及P-YAP蛋白表达水平降低;与模型组比较,活血解毒方组大鼠视网膜中P-MST及P-YAP蛋白表达水平升高。Western blot的结果显示:与正常组比较,模型组大鼠视网膜中Lats1、TAZ及TEAD蛋白表达水平升高;与模型组比较,活血解毒方组大鼠视网膜中Lats1、TAZ及TEAD蛋白表达水平降低。结论活血解毒方可能通过Hippo信号通路改善糖尿病视网膜病变。
Objective To observe the effect of Huoxue Jiedu Fang on Hippo signal pathway in the retina of diabetic rats. Methods Twenty-four healthy SD rats were selected and fasted for 12 h. Diabetic rats were induced by a single intraperitoneal injection of streptozotocin (65 mg / kg). After 20 weeks of modeling, rats were randomly divided into model group and Huoxue Jiedu Fang group. Another 10 rats were selected as normal control group, given equal volume of distilled water gavage. Each group intervention once a day for a total of 12 weeks. Immunohistochemistry and Western blot were used to detect phosphorylated mammalian male sterile line 20-like kinase (P-MST), large tumor suppressor gene 1 (Lats1), phosphorylated Yes-related protein (P-YAP), transcriptional factor (TAZ) and TEA (TEAD). Results The results of immunohistochemistry showed that the expression of P-MST and P-YAP in retina of model group decreased compared with normal group. Compared with model group, the expression of P-MST and P -YAP protein expression increased. The results of Western blot showed that compared with the normal group, the expression of Lats1, TAZ and TEAD protein in retina of the model group increased; compared with the model group, the expression of Lats1, TAZ and TEAD protein in the retina of the Huoxue Jiedu Decoction group decreased . Conclusion Huoxue Jiedu Fang may improve diabetic retinopathy through Hippo signaling pathway.