Wolf W C, Yoshida H, Agata J, Chao L, Chao J
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425-2211, USA.
Kidney Int. 2000 Aug;58(2):730-9. doi: 10.1046/j.1523-1755.2000.00219.x.
Tissue kallikrein cleaves kininogen substrate to produce the potent vasodilating peptide kinin, which plays important roles in cardiovascular and renal function. To explore cardiac and renal potential protective effects of kallikrein gene delivery in chronic renal failure, we delivered adenovirus carrying the human tissue kallikrein cDNA (cHK) into rats with 5/6 reduction of renal mass.
Expression of human tissue kallikrein in rats was assessed by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR)/Southern blotting. Physiological parameters monitored in rats included systolic blood pressure, heart rate, and urinary excretion of protein, albumin, kinin, cGMP, cAMP, and nitrate/nitrites. Systemic and regional hemodynamics were measured by fluorescent-labeled microspheres. Heart weight and myocyte diameter were used to assess left ventricular hypertrophy. Quantitative and qualitative morphological analyses were used to evaluate histologic changes in kidney and heart sections.
Active tissue kallikrein reached a peak serum level of 463 +/- 76 ng/mL following gene delivery and returned to control levels within 21 days. A maximal blood pressure reduction of 37 mm Hg was observed within one week in rats receiving kallikrein gene delivery as compared with control rats receiving adenovirus containing the luciferase gene (159 +/- 5 vs. 196 +/- 6 mm Hg, N = 15, P < 0.001), and a significant blood pressure difference continued for five weeks postgene delivery. Kallikrein gene delivery significantly decreased total urinary protein and albumin excretion and increased levels of urinary kinin, nitrite/nitrate, and cGMP levels. Cardiac output and regional blood flow were also increased, while peripheral vascular resistance decreased. Kallikrein gene transfer reduced glomerular sclerotic lesions, tubular damage, lumenal protein cast accumulation, and interstitial inflammation in the kidney. Myocardial hypertrophy and fibrosis were also attenuated in rats receiving kallikrein gene delivery.
These findings indicated that kallikrein gene delivery attenuates hypertension and protects against renal injury and cardiac remodeling in the rat remnant kidney model of chronic renal failure.
组织激肽释放酶可裂解激肽原底物以产生强效血管舒张肽激肽，其在心血管和肾功能中发挥重要作用。为了探究激肽释放酶基因递送对慢性肾衰竭心脏和肾脏的潜在保护作用，我们将携带人组织激肽释放酶cDNA（cHK）的腺病毒导入肾质量减少5/6的大鼠体内。
通过酶联免疫吸附测定（ELISA）以及逆转录-聚合酶链反应（RT-PCR）/Southern印迹法评估大鼠中人组织激肽释放酶的表达。监测大鼠的生理参数包括收缩压、心率以及蛋白质、白蛋白、激肽、环磷酸鸟苷（cGMP）、环磷酸腺苷（cAMP）和硝酸盐/亚硝酸盐的尿排泄量。通过荧光标记微球测量全身和局部血流动力学。心脏重量和心肌细胞直径用于评估左心室肥大。采用定量和定性形态学分析评估肾脏和心脏切片的组织学变化。
基因递送后，活性组织激肽释放酶血清水平达到峰值463±7（6）ng/mL，并在21天内恢复至对照水平。与接受含荧光素酶基因腺病毒的对照大鼠相比，接受激肽释放酶基因递送的大鼠在一周内观察到最大血压降低37 mmHg（159±5 vs. 196±6 mmHg，N = 15，P < 0.001），并且在基因递送后五周内血压仍存在显著差异。激肽释放酶基因递送显著降低尿总蛋白和白蛋白排泄量，并增加尿激肽、亚硝酸盐/硝酸盐和cGMP水平。心输出量和局部血流量也增加，而外周血管阻力降低。激肽释放酶基因转移减少了肾脏中的肾小球硬化病变、肾小管损伤、管腔内蛋白管型积聚和间质炎症。接受激肽释放酶基因递送的大鼠心肌肥大和纤维化也得到减轻。
这些发现表明，在慢性肾衰竭大鼠残余肾模型中，激肽释放酶基因递送可减轻高血压，并预防肾损伤和心脏重塑。
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