摘要
目的探讨瞬时受体电位阳离子通道蛋白6(TRPC6)在正常人、小鼠和大鼠肾组织及小鼠足细胞系(MPC5)的表达和分布,为研究TRPC6在肾脏的功能及其与足细胞分子间的关系奠定基础。方法1.应用免疫组织化学方法观察TRPC6在正常人、小鼠和大鼠肾组织及MPC5分布。2.通过反转录酶-聚合酶链反应(RT-PCR)检测TRPC6在小鼠肾组织和MPC5表达。3.应用免疫蛋白印迹检测TRPC6在正常人、小鼠和MPC5表达。结果1.TRPC6在正常人肾小球呈弱表达,在肾小管和肾血管表达较强,在小鼠和大鼠主要沿肾小球毛细血管袢和系膜区分布,肾间质也有少许表达。TRPC6在MPC5的荧光染色为阳性,在分化态细胞主要分布于胞膜表面。2.在小鼠肾组织及MPC5均检测到特异性TRPC6的PCR产物条带。3.在正常人、小鼠肾组织及MPC5均检测到特异性的相对分子质量为106的TRPC6蛋白条带。结论TRPC6在正常人、小鼠和大鼠肾小球均有表达,在mRNA及蛋白水平均证实MPC5能表达TRPC6,为从离子通道的角度利用MPC5探讨蛋白尿发生的分子机制奠定基础。
Objective To explore the expression and distribution of transient receptor potential cation channel 6（TRPC6）in normal human,mice,rats＇ renal tissue and the mouse podocyte clone 5（MPC5）for further investigating the relationship between TRPC6 and the protei-nuria-related podocyte molecules.Methods 1.The distributions of TRPC6 in normal human,mice,rats＇ renal tissue and MPC5 were observed by using the immunochemistry staining.2.The mRNA expression of TRPC6 in mouse renal cortex and differentiated MPC5 was detected by using reverse transcriptase-polymerase chain reaction（RT-PCR）.3.The protein expression of TRPC6 in human,mice and differentiated MPC5 was detected by using Western blotting.Results 1.In human kidney,TRPC6 showed a weak staining in glomeruli and a strong staining in renal tubules and vessels.In mice and rats＇ kidney,TRPC6 showed a strong staining in glomeruli and was mainly distributed along the capillary loops of glomerulus and in mesangium.The positive staining of TRPC6 was observed in MPC5,which was distributed evenly on the cell membrane in differentiated podocytes.2.The specific PCR band of TRPC6 was detected in mouse renal cortex and differentiated MPC5.3.The specific protein band of TRPC6 was detected in normal human,mice renal cortex and differentiated MPC5 with the size of 106.Conclusions The expression of TRPC6 is verified in normal human,mice and rats＇ kidneys,and in differentiated MPC5.These results will benefit for further exploring the relationship between TRPC6 and the proteinuria-related podocyte molecules.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2007年第17期1293-1295,共3页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金项目资助(30170992
30672259)
北京市自然基金项目资助(7072080)
北京大学第一医院科研基金项目资助
关键词
肾脏
足细胞
瞬时受体电位阳离子通道蛋白6
表达
分布
kidney
podocyte
transient receptor potential cation channel 6
expression
distribution
