牡蛎提取物对足细胞裂孔膜蛋白NEPHRIN和ERK通路的影响  

Effect of Oyster Extract on NEPHRIN Protein and ERK Pathway

作　　者：王竹[1] 孙万森[1] 李睿萍[1] 王晔[1] 刘军花[1] 

机构地区：[1]西安交通大学医学院第二附属医院,陕西西安710004

出　　处：《中医药导报》2015年第23期7-12,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：陕西省中医药管理局项目(jc38.2011-2014);项目名称:牡蛎提取物对阿霉素肾病大鼠足细胞骨架及裂孔隔膜相关蛋白;信号通路影响的实验研究

摘　　要：目的:探讨牡蛎提取物对足细胞裂孔膜蛋白NEPHRIN细胞外调节蛋白激酶(ERK)通路的影响。方法:体内实验采用一次性尾静脉注射阿霉素(adriamycin,ADR)的方法,建立阿霉素肾病大鼠模型。随机分为空白对照组、模型对照组、牡蛎提取物小、中、大剂量组和阳性对照组。从第4周起,牡蛎提取物组分别给予牡蛎提取物3.0 g/kg、6.0 g/kg、12.0 g/kg灌胃,阳性对照组给予泼尼松25 mg/kg灌胃。连续28 d后,进行24 h尿蛋白定量、血清生化指标检测,实验结束时用Real-time PCR法及Western Blot法检测足细胞骨架蛋白NEPHRIN m RNA及NEPHRIN蛋白表达。体外选用1μmol/L阿霉素制造足细胞损伤模型,Western Blot检测NEPHRIN蛋白含量及足细胞ERK信号通路的磷酸化水平。结果:牡蛎提取物可升高血浆白蛋白、降低总胆固醇的含量,同时减少尿蛋白;阿霉素肾病大鼠及阿霉素诱导的足细胞中NEPHRIN蛋白表达均下降。经牡蛎提取物干预后,可以上调体内、体外的NEPHRIN蛋白表达;阿霉素(1μmol/L)可显著升高ERK的磷酸化水平,与空白对照组比较,差异有统计学意义(P<0.05)。牡蛎提取物预处理细胞后,能够有效地遏制阿霉素诱导的ERK通路活化,与模型对照组比较,差异有统计学意义(P<0.05)。结论:牡蛎提取物对足细胞裂孔膜蛋白NEPHRIN有调控作用,可能是通过抑制ERK通路活化而实现的。Objective： To investigate the effect of oyster extract on podocyte protein NEPHRIN and ERK pathway. Methods： In vivo by a single intravenous injection of adriamycin （adriamycin, ADR） method, the rat model of adriamycin induced nephropathy rats was established. The rats were randomly divided into blank control group, model control group, oyster extract small, high dose group and positive control group. From the fourth week, treatment groups were given the oyster, oyster extract 3 g/kg, 6 g/kg and 12 g/kg orally respectively,and the positive control group was given prednisone 25 mg/kg orally. For 28 consecutive days, 24 hours urine protein and serum biochemical indexes were detected; at the end of the experiment, Real-time PCR method and Western Blot method was used to detect the expression of podocyte cytoskeleton protein NEPHRIN mRNA and NEPHRIN protein. In vitro selection, 1 μmol/L doxorubicin was used to create podocyte injury model,and Western Blot was applied to detect the NEPHRIN protein content and ERK of podocyte signaling pathway phosphorylation. Results： The oyster extract can increase the plasma albumin and reduce total cholesterol as well as urinary protein. Expression of NEPHRIN protein was decreased in podocyte injury model created by doxorubicin. In adriamycin nephropathy rats and adriamycin induced in podocytes, the oyster extract intervention can increase the expression of NEPHRIN protein in vivo and in vitro. Adriamycin （1 μmol/L） significantly increased ERK phosphorylation level,and there were significant difference between the control group and blank control（P〈0.05）. Pretreatment of cells with oyster extract, can effectively prevent the adriamycin induced activation of ERK pathway,and there were significant difference between the control group and the model （P〈0.05）. Conclusion： The oyster extract has a role in the regulation of podocyte slit diaphragm protein NEPHRIN, which may be achieved by inhibiting the activation ofthe ERK collaterals.

关 键 词：牡蛎提取物 NEPHRIN ERK信号通路 

分 类 号：R285.5[医药卫生—中药学]