解毒化瘀方药物血清与药物血浆对模型PC12细胞蛋白酶活化受体―1细胞外信号调节激酶12表达影响.docVIP

解毒化瘀方药物血清与药物血浆对模型PC12细胞蛋白酶活化受体―1细胞外信号调节激酶12表达影响 　　摘要：目的 比较解毒化瘀方药物血清与药物血浆对凝血酶合并缺氧诱导PC12细胞损伤蛋白酶活化受体（PAR）-1、细胞外信号调节激酶（ERK）1/2表达的差异。方法 灌胃给药制备大鼠药物血清和药物血浆。采用三气培养箱和无糖DMEM培养液造成细胞缺氧模拟缺血，并同时加入150 U/mL凝血酶，建立体外缺氧合并凝血酶诱导的PC12细胞损伤模型。实验分为空白组、模型组、药物血清组及药物血浆组。实时荧光定量PCR检测细胞PAR-1、ERK1/2 mRNA表达，免疫荧光法检测细胞PAR-1、ERK1/2蛋白表达。结果 与空白组比较，模型组PAR-1、ERK1/2 mRNA和蛋白表达增强（P0.05）；与模型组比较，药物血清组和药物血浆组PAR-1、ERK1/2 mRNA和蛋白表达下降（P0.05，P0.01）；药物血浆组PAR-1、ERK1/2表达较药物血清组下降（P0.05）。结论 解毒化瘀方药物血浆抗缺氧合并凝血酶诱导PC12细胞损伤作用优于其药物血清。 　　关键词：解毒化瘀方；药物血浆；药物血清；PC12细胞；大鼠 　　DOI：10.3969/j.issn.1005-5304.2016.06.019 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）06-0073-04 　　Abstract： Objective To compare the differences between Jiedu Huayu Prescription containing serum and plasma on PAR-1， ERK1/2 expression of the PC12 cell injury model. Methods Rat serum containing and plasma models was prepared through gavage. Three gas incubators and sugar-free DMEM medium simulated ischemia caused by hypoxia were used， and 150 U/mL of thrombin was added， with a purpose to establish the thrombin-induced hypoxic PC12 cell injury model. The experiment was divided as control group， model group， serum containing group and plasma containing group. Real-time fluorescent quantitative PCR was used to detect the expression levels of PAR-1 and ERK1/2 mRNA in each group； immunofluorescence was used to detect the protein expressions of PAR-1 and ERK1/2. Results Compared with the control group， the expressions of PAR-1 and ERK1/2 mRNA and protein in model group were enhanced （P0.05）； compared with the model group， the expressions of PAR-1 and ERK1/2 mRNA and protein in serum containing group and plasma containing group decreased （P0.05， P0.01）； the expressions of PAR-1 and ERK1/2 mRNA and protein in plasma containing group decreased more significantly than serum containing group （P0.05）. Conclusion Jiedu Huayu Prescription containing plasma was superior to its containing serum against the injury of PC12 cell induced by hypoxic and thrombin. 　　Key words： Jie