摘要
通过化瘀通络中药对H-2Kb-ts A58条件性永生化小鼠足细胞裂孔膜蛋白podocin、CD2AP表达的干预作用,探讨化瘀通络中药降低糖尿病肾病蛋白尿、保护肾功能的可能作用机制。实验以体外培养小鼠足细胞为研究对象,将其分为正常糖组(CG)、高糖组(HG)、高糖加药组(HG+Z),通过免疫细胞化学(ICC)、蛋白质印迹法(Western-Blot)、实时荧光定量(Real-time PCR)方法检测不同时间点不同干预后各组podocin、CD2AP蛋白及基因的表达情况。结果显示干预足细胞48 h后,与CG组比较,其余两组podocin、CD2AP两种蛋白表达量均明显降低(P<0.05,P<0.01);与HG组比较,(HG+Z)组两种蛋白表达量均明显升高(P<0.05)。与同时间点CG组比较,其余两组podocin、CD2AP mRNA表达均显著降低(P<0.01);与同时间点HG组比较,(HG+Z)组48 h两种蛋白的基因表达量均明显升高(P<0.01),24、72 h与同时间点HG组无明显差异(P>0.05)。以上结果说明化瘀通络中药可以上调高糖刺激下足细胞裂孔膜蛋白podocin、CD2AP的表达,其可能为化瘀通络中药减少糖尿病肾病蛋白尿、保护肾功能的作用机制之一。
To explore the possible mechanism of Huayu Tongluo traditional Chinese medicine in reducing proteinuria and protecting renal function of diabetic nephropathy on expression of podocin, CD2AP in the conditionally immortalized mouse podoeytes. The conditionally immortalized mouse podocytes were selected as the research object. These podocytes cultured under the growth permissive condition were divided into the normal glucose group （ CG group）, high glucose group （ HG group） and high glucose dosing group （ HG ＋ Z group） and intervened for 24 h,48 h and 72 hours. The ex- pression of podocin and CD2AP was detected by ICC, Western-blot and Real-Time PCR. It was observed that at 48 hours ,compared with CG group,the expression of podoein and CD2AP protein in HG and （HG ＋ Z） groups was lower （ P 〈 0.05, P 〈 0.01 ）. Compared with HG group, the expression of podocin and CD2AP protein in （ HG ＋ Z） group was higher （P 〈 0.05）. Compared with CG group, at same time point, the expression of podocin and CD2AP mRNA in other groups was lower （P 〈 0.01 ）. Compared with HG group, only at 48 hours, the expression of podocin and CD2AP mRNA in （HG ＋ Z） group was higher （P 〈0.01 ）. The results indicated that Huayu Tongluo traditional Chinese medicine can increase the expression of membrane protein podocin, CD2AP and it may be one of the mechanisms on reducing diabetic nephropathy proteinuria and protecting renal function.
出处
《天然产物研究与开发》
CAS
CSCD
北大核心
2016年第12期1886-1890,共5页
Natural Product Research and Development
基金
国家自然科学基金(81373804)
