Sebeková K, Dämmrich J, Krivosíková Z, Heidland A
Clinic of Pharmacotherapy, Institute of Preventive and Clinical Medicine, Bratislava, Slovakia.
Res Exp Med (Berl). 1999 Dec;199(3):177-88. doi: 10.1007/s004330050122.
It has been demonstrated that intraperitoneal administration of proteolytic enzymes ameliorates the progression of renal diseases in various animal models. In the present study, we employed the rat remnant kidney model to study the effectiveness of oral administration of proteases. Twenty male Wistar rats underwent sham operation (CTRL), while 25 were subjected to 5/6 nephrectomy (5/6 NX). Rats were randomised into placebo (PL) (2 ml tap water/day by gavage), or Phlogenzym (E; fixed mixture of trypsin 2.42 mg, bromelain 4.54 mg, and rutozid 5.04 mg added as antioxidant, in 2 ml tap water daily by gavage) treated group. Duration of the study was 45 days. Rats were pair-fed. Enzyme treatment exerted salutary effects on various functional and morphological parameters. Proteinuria was higher in both 5/6 NX group rats throughout the study. Administration of proteases ameliorated its rise effectively (data at sacrifice: CTRL-PL 6.27 +/- 1.25, CTRL-E 9.27 +/- 0.99, 5/6 NX-PL 74.04 +/- 21.33, 5/6 NX-E 39.09 +/- 7.93 mg/24 h; P < 0.01). Increased urinary excretion of the fibrogenic cytokine transforming growth factor (TGF-beta 1) was improved, too (CTRL-PL 0.349 +/- 0.051, CTRL-E 0.693 +/- 0.230, 5/6 NX-PL 3.044 +/- 0.540, 5/6 NX-E 1.390 +/- 0.238 ng/mumol creatinine; P < 0.05). At sacrifice, tubulointerstitial fibrosis was less pronounced in E-treated rats. Correspondingly, the volume fraction of tubulointerstitial tissue in the renal cortex was improved in 5/6 NX-E rats (CTRL-PL 9.9 +/- 0.2, CTRL-E 10.0 +/- 0.2, 5/6 NX-PL 17.9 +/- 1.8, 5/6 NX-E 13.8 +/- 0.9%; P < 0.05). The protein/DNA ratio in isolated glomeruli and tubules, as an estimate of glomerular matrix accumulation and hypertrophy of tubules, was enhanced in 5/6 NX groups and a tendency towards lower values was observed after E treatment. Renal function as evaluated by serum creatinine and urea levels was not influenced by the enzyme therapy. No between-group differences in blood pressure were observed. In summary, oral administration of proteolytic enzymes improved proteinuria and urinary TGF-beta 1 excretion, as well as the severity of tubulointerstitial fibrosis without signs of toxicity.
已证明腹腔注射蛋白水解酶可改善多种动物模型中肾脏疾病的进展。在本研究中，我们采用大鼠残肾模型来研究口服蛋白酶的效果。20只雄性Wistar大鼠接受假手术（CTRL），而25只大鼠接受5/6肾切除术（5/6 NX）。大鼠被随机分为安慰剂（PL）组（每天经口灌胃2 ml自来水）或菠萝蛋白酶（E）组（在2 ml自来水中添加2.42 mg胰蛋白酶、4.54 mg菠萝蛋白酶和5.04 mg芦丁作为抗氧化剂，每天经口灌胃）。研究持续时间为45天。大鼠采用配对饲养。酶治疗对各种功能和形态学参数产生有益影响。在整个研究过程中，5/6 NX组大鼠的蛋白尿均较高。蛋白酶给药有效改善了蛋白尿的增加（处死时数据：CTRL-PL 6.27±1.25，CTRL-E 9.27±0.99，5/6 NX-PL 74.04±21.33，5/6 NX-E 39.09±7.93 mg/24 h；P<0.01）。促纤维化细胞因子转化生长因子（TGF-β1）的尿排泄增加也得到改善（CTRL-PL 0.349±0.051，CTRL-E 0.693±0.230，5/6 NX-PL 3.044±0.540，5/6 NX-E 1.390±0.238 ng/μmol肌酐；P<0.05）。处死时，E治疗组大鼠的肾小管间质纤维化不那么明显。相应地，5/6 NX-E组大鼠肾皮质肾小管间质组织的体积分数得到改善（CTRL-PL 9.9±0.2，CTRL-E 10.0±0.2，5/6 NX-PL 17.9±1.8，5/6 NX-E 13.8±0.9%；P<0.05）。5/6 NX组中，分离的肾小球和肾小管中的蛋白质/DNA比值升高，这是肾小球基质积聚和肾小管肥大的一个指标，E治疗后观察到该比值有降低的趋势。通过血清肌酐和尿素水平评估的肾功能不受酶治疗的影响。未观察到组间血压差异。总之，口服蛋白水解酶可改善蛋白尿和尿TGF-β1排泄，以及肾小管间质纤维化的严重程度，且无毒性迹象。
