复方肾华片对缺氧/复氧肾小管上皮细胞NaDC1转运蛋白定位的影响

The Effects of Shenhua Tablet Compound on the Location of NaDC1 in Renal Tubular Epithelial Cells with Anoxia /reoxygenation

目的:观察复方肾华含药血清对缺氧/复氧模型肾小管上皮细胞钠/二羧酸协同转运蛋白1(NaDC1)的定位影响。方法:雄性wistar大鼠,灌胃给药3 d后处死,分离含药血清备用。分离培养大鼠肾小管上皮细胞,随机分4组:正常血清GFP空质粒组、正常血清GFP-NaDC1质粒组、肾华含药血清GFP空质粒组和肾华含药血清GFP-NaDC1质粒组,分别进行质粒转染后,在正常培养基或肾华含药血清培养基中培养,进行缺氧/复氧处理后应用免疫荧光和生物素化免疫印迹技术观察肾小管上皮细胞NaDC1的分布变化及表达。结果:免疫荧光结果显示,正常血清GFP空质粒组、正常血清GFP-NaDC1质粒组和肾华含药血清GFP空质粒组NaDC1均匀分布在细胞内,肾华含药血清GFP-NaDC1质粒组NaDC1主要分布于胞膜。免疫印迹结果显示肾华含药血清GFP-NaDC1质粒组NaDC1顶端分布明显多于基底端,其余各组细胞NaDC1在顶端和基底端分布差异无统计学意义。结论:复方肾华含药血清可以有效促进缺氧/复氧损伤大鼠肾小管上皮细胞极性修复。

Objective： To observe the effect of the Shenhua - containing serum on the location of Sodium - dependent dicar- boxylate transporter 1 （ NaDC1 ） in the renal tubular epithelial cells of rats. Methods： After 3 days of continuous intragastric drug ad- ministration, male Wistar rats were sacrificed and Shenhua - containing serum has been prepared and separated. Renal tubular epithe- lial cells （RTECs） were divided into a normal serum- GFP- empty plasmid group, a normal serum- GFP- NaDC1 plasmid group, a Shenhua -containing serum -GFP- empty plasmid group and a Shcnhua -containing serum -GFP- NaDC1 plasmid group. After anoxia/reoxygenation, irarnunofluorescence and immunoblot were performed. Results： Immunofluorescence examinations revealed a widened distribution of NaDC1 in normal serum- GFP- empty plasmid group, normal serum- GFP- NaDC1 plasmid group and Shenhua - containing serum - GFP - empty plasmid group, in Shenhua - containing serum - GFP - NaDC1 plasmid group, NaDC1 was distributed at membrane. Immunoblot show that approximately 90% of the NaDC1 was expressed at the basolateral membrane of RTECs in Shenhua - containing serum - GFP - NaDC1 plasmid group, whereas other three groups were expressed on the apical and basolateral surfaces at the nearly equal level. Conclusion：The Shenhua -containing serum effectively promoted the recovery of the po- larity of RTECs caused by anoxia/reoxygenation.