摘要
目的探讨肾康丸含药血清对晚期氧化蛋白产物(AOPP)诱导的体外培养足细胞(MPC5)单核细胞趋化蛋白1(MCP-1)表达的影响及其机制。方法制备肾康丸含药血清和AOPP;体外培养MPC5,用不同浓度的肾康丸含药血清刺激不同时间,MTT法检测细胞增殖;用肾康丸含药血清干预AOPP处理的MPC5,Western blot法检测p38MAPK、IκBα蛋白表达,免疫荧光法观察NF-κB p65核转移,ELISA法检测细胞培养上清中MCP-1表达。结果 (1)肾康丸含药血清呈时间和浓度依赖性促进MPC5增殖,最佳作用浓度和时间分别是5%、24 h;(2)AOPP呈浓度和时间依赖性诱导MPC5分泌MCP-1;p38抑制剂SB203580或NF-κB抑制剂小白菊内酯预孵育能使细胞上清中MCP-1分泌减少;肾康丸含药血清能抑制AOPP诱导MPC5分泌MCP-1;(3)AOPP以浓度依赖的方式增加P-p38蛋白的表达、降低IκBα蛋白的表达;与AOPP组相比,AOPP+肾康丸组MPC5 IκBα蛋白表达增加;AOPP+SB203580组IκBα蛋白表达亦增加,但不及AOPP+肾康丸组;(4)肾康丸含药血清减少AOPP诱导的NF-κB p65核转移。结论肾康丸含药血清能通过p38MAPK/NF-κB途径下调MCP-1表达而发挥抗炎作用,为应用其防治糖尿病肾病提供了新的理论依据。
Objective To investigate the effect of the serum of rats fed with Shenkang pill in regulating monocyte chemoattractant protein I （MCP-1） expression induced by advanced oxidation protein products （AOPP） in mouse podocyte clone 5 （MPC5） and explore the underlying mechanism. Methods MPC5 cultured in vitro were incubated for different time lengths in the presence of different concentrations of serum of rats medicated with Shenkang pill, and the cell proliferation was assessed using MTT assay. In MPC5 treated with AOPP prior to exposure to the rat serum, the changes in the protein expressions of p38MAPK and IlcBa were examined with Western blotting, NF-lcB p65 nuclear translocation was analyzed with immunofluorescence assay, and MCP-1 expression in the supernatant was determined using ELISA kits. Results The medicated rat serum time- and concentration-dependently promoted the proliferation of MPCS, with the optimal serum concentration of 5% and incubation time of 24 h. AOPP significantly increased MCP-1 expression in the cell supernatant in a time-and concentration-dependent manner; pretreatment with SB203580 （a p38 inhibitor） or parthenolide （a NF-nB inhibitor） significantly decreased MCP-1 expression, and treatment with the medicated serum significantly decreased AOPP-induced MCP-1 expression. AOPP concentration-dependently increased the protein expression of P-p38 but decreased that of IkBa. Both the medicated serum and SB203580 increased IkBa protein in AOPP-induced cells, but the effect was more obvious with the medicated serum. The medicated serum also decreased NF-kB p65 nuclear translocation in AOPP-induced MPC5. Conclusion Shenkang pill-medicated serum can decrease AOPP-induced expression of MPC-1 in MPC5 by regulating p38MAPK/ NF-kB to mediate its anti-inflammatory effect. This finding provides a new theoretical basis for the application of Shenkang pill to treat diabetic nephropathy.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第9期1265-1271,共7页
Journal of Southern Medical University
基金
国家自然科学基金(81072848)
国家级大学生创新创业训练计划项目(201212121021)
广东省科技计划项目(2011B061300014)~~
关键词
肾康丸
晚期氧化蛋白产物
足细胞
单核细胞趋化蛋白1
P38MAPK
NF-ΚB
Shenkang pill
advanced oxidation protein products
mouse podocyte clone 5
monocyte chemoattractant protein 1
p38MAPK
nuclear factor-kB
