Chen M, Harris M P, Rose D, Smart A, He X R, Kretzler M, Briggs J P, Schnermann J
Department of Physiology, University of Michigan, Ann Arbor 48109.
J Clin Invest. 1994 Jul;94(1):237-43. doi: 10.1172/JCI117312.
The present study was undertaken to assess the presence of renin enzymatic activity and renin mRNA in proximal tubules of rat kidneys, and to determine the effect of converting enzyme inhibition (CEI) on proximal tubule renin gene expression. Proximal convoluted tubules (PCT), proximal straight tubules (PST), outer medullary collecting ducts (OMCD), and glomeruli (Gloms) were isolated by microdissection. Renin activity was measured in sonicated segments by radioimmunoassay. Renin mRNA levels were assessed using a quantitative PCR. Renin activity in PCT averaged 51 +/- 15 microGU/mm compared to 405 +/- 120 microGU/glomerulus. No measurable renin activity was found in PST and OMCD. Renin activity in both glomeruli and tubules had the same pH optimum, between 7.0 and 7.5. Renin mRNA was consistently detectable in cDNA prepared from PCT and PST, although its abundance per mm tubule was about 1/500th that found in one glomerulus. Renin mRNA was not detectable in OMCD. Tubular renin PCR product identity was confirmed by restriction digestion. CEI administration increased glomerular renin activity and renin mRNA, but not proximal tubular renin. The absence of a stimulatory effect of CEI on proximal tubule renin gene expression suggests the operation of different intracellular signals in control of renin synthesis in the proximal tubule than in the vascular compartment.
本研究旨在评估大鼠肾脏近端小管中肾素酶活性和肾素mRNA的存在情况,并确定转换酶抑制(CEI)对近端小管肾素基因表达的影响。通过显微切割分离出近端曲管(PCT)、近端直小管(PST)、外髓集合管(OMCD)和肾小球(Gloms)。通过放射免疫测定法测量超声处理后的节段中的肾素活性。使用定量PCR评估肾素mRNA水平。PCT中的肾素活性平均为51±15微GU/mm,而肾小球中的肾素活性为405±120微GU/肾小球。在PST和OMCD中未发现可测量的肾素活性。肾小球和小管中的肾素活性在pH值7.0至7.5之间具有相同的最佳值。从PCT和PST制备的cDNA中始终可检测到肾素mRNA,尽管每毫米小管中的丰度约为一个肾小球中的1/500。在OMCD中未检测到肾素mRNA。通过限制性消化确认了肾小管肾素PCR产物的同一性。给予CEI可增加肾小球肾素活性和肾素mRNA,但不增加近端小管肾素。CEI对近端小管肾素基因表达无刺激作用,这表明在近端小管和血管腔室中控制肾素合成的细胞内信号不同。