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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 extracellular regulated protein kinase (ERK) pathway of podocyte mesoporous membrane protein NEPHRIN. Methods: The rat model of adriamycin nephropathy was established by injecting adriamycin (ADR) into the tail vein in vivo. Randomly divided into blank control group, model control group, oyster extract small, medium and large dose groups and positive control group. From the 4th week onwards, oyster extract group were orally administered 3.0 g / kg, 6.0 g / kg, and 12.0 g / kg orally respectively. The positive control group was given prednisone 25 mg / kg orally. After 28 days, 24 h urinary protein and serum biochemical indexes were measured. At the end of experiment, the expressions of NEPHRIN m RNA and NEPHRIN protein were detected by Real-time PCR and Western Blot. 1μmol / L doxorubicin was used in vitro to establish the model of podocyte injury. Western Blot was used to detect the protein level of NEPHRIN and phosphorylation of p ERK signaling pathway. Results: Oyster extract increased plasma albumin, decreased total cholesterol and reduced urinary protein. NEPHRIN protein expression in doxorubicin-induced nephropathy rats and doxorubicin-induced podocytes decreased. The intervention of oyster extract could up-regulate NEPHRIN protein expression in vitro and in vivo. Adriamycin (1μmol / L) significantly increased the phosphorylation of ERK, which was significantly different from that of the blank control group (P <0.05) ). Oyster extract pretreatment cells, can effectively curb adriamycin-induced ERK pathway activation, compared with the model control group, the difference was statistically significant (P <0.05). Conclusion: Oyster extract can regulate the podocyte membrane pore membrane protein NEPHRIN, which may be through inhibiting the activation of ERK pathway.