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作者：高洁福建中医学院

学位级别：硕士

目的:本课题旨在通过动物实验,研究桃核真武汤与益肾降浊汤对5/6肾切除所致慢性肾功能衰竭大鼠肾小管功能的影响,从细胞分子水平观察两方防治慢性肾功能衰竭的作用机理。方法:采用5/6肾切除所致慢性肾衰大鼠作为研究对象,SD大鼠78只,雌... 详细信息

目的:本课题旨在通过动物实验,研究桃核真武汤与益肾降浊汤对5/6肾切除所致慢性肾功能衰竭大鼠肾小管功能的影响,从细胞分子水平观察两方防治慢性肾功能衰竭的作用机理。方法:采用5/6肾切除所致慢性肾衰大鼠作为研究对象,SD大鼠78只,雌雄各半,随机分为空白对照组8只、假手术组10只和手术组60只。手术组采用两步法制备5/6肾切除CRF模型后,再随机分为病理模型组、依那普利对照组、大黄颗粒对照组、益肾降浊治疗组、桃核真武治疗组,手术四周后,开始六周的治疗。观察治疗后大鼠的一般状况,治疗结束后测定各组大鼠的血清BUN、Scr、β2-MG及尿β2-MG、RBP、NAG酶;同时取上述各组大鼠肾脏观察其形态学改变。结果:1.病理模型组大鼠血清BUN、Scr、β2-MG和尿β2-MG、RBP、NAG酶水平显著高于正常组（P＜0.01）,说明5/6肾切除大鼠模型存在肾小管功能的损伤。2.桃核真武汤和益肾降浊汤能明显改善5/6肾切除大鼠的一般情况,降低血清BUN、Scr、β2-MG和尿β2-MG、RBP、NAG酶水平（P＜0.01）,光镜下显示两方可以改善肾小管-间质病理损伤,效果优于大黄颗粒和依那普利。3.治疗组中桃核真武汤和益肾降浊汤比较,二者无明显差异。结论:桃核真武汤与益肾降浊汤能有效降低5/6肾切除大鼠慢性肾衰模型血清BUN、Scr、β2-MG和尿β2-MG、RBP、NAG酶,一定程度上改善肾小管-间质的病理损伤,对大鼠肾切除所致慢性肾衰肾小管-间质损伤的改善有一定的疗效,效果优于依那普利和大黄颗粒;桃核真武汤与益肾降浊汤是防止或延缓肾小管-间质损害至终末期肾衰的有效方剂。

关键词：肾功能衰竭,慢性/中医药疗法肾小管/药物作用复方/药理学经方/药理学动物,实验 @桃核真武汤 @益肾降浊汤

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Influence of exogenous thiols on inorganic mercury-induced injury in renal proximal and distal tubular cells from normal and uninephrectomized rats

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第2期291卷 492-502页

作者： Lash, LH Putt, DA Zalups, RK Wayne State Univ Sch Med Dept Pharmacol Detroit MI 48201 USA Mercer Univ Sch Med Div Basic Med Sci Macon GA 31207 USA

Inorganic mercury (Hg2+) induced time- and concentration-dependent cellular injury in freshly isolated proximal tubular (PT) and distal tubular (DT) cells from normal (control) rats or uninephrectomized (NPX) rats. PT cells from NPX rats were more susceptible than PT cells from control rats, and DT cells were slightly more susceptible than PT cells to cellular injury induced by Hg2+ (not bound to a thiol). Preloading cells with glutathione increased Hg2+-induced cellular injury in PT cells from control rats. However, coincubation of PT or DT cells from control or NPX rats with Hg2+ and glutathione (1:4) provided significant protection relative to incubations with Hg2+ alone. No support was obtained for a role for gamma-glutamyltransferase in glutathione-dependent protection. However, the organic anion carrier does appear to play a role in accumulation and toxicity of mercuric conjugates of cysteine in PT cells from control, but not NPX, rats. Coincubation with Hg2+ and cysteine (1:4) had little effect on, or slightly enhanced, Hg2+-induced cellular injury at low concentrations of Hg2+ in all cells studied. Coincubation with Hg2+ and albumin (1:4) markedly protected PT and DT cells from control and NPX rats at all concentrations except the highest concentration of Hg2+ in DT cells from NPX rats. 2,3-Dimercapto-1-propanesulfonic acid protected cells both when preloaded or added simultaneously with Hg2+. Thus, renal cells from NPX rats are more susceptible to Hg2+-induced injury, PT and DT cells respond differently to exposure to Hg2+, and thiols can significantly modulate the toxic response to Hg2+.

关键词：白蛋白类/药理学半胱氨酸/药理学剂量效应关系药物药物相互作用谷胱甘肽/药理学无机化学品/毒性肾小管/药物作用肾小管远端/药物作用肾小管近端/药物作用汞/毒性大鼠 Sprague-Dawley 时间因素二巯丙磺钠/药理学 γ-谷氨酰转移酶/生理学动物男(雄)性大鼠

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Effects of β-adrenergic stimulation on the acutely obstructed ureter in dogs

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第1期292卷 67-75页

作者： Murakami, M Tomiyama, Y Hayakawa, K Akahane, M Ajisawa, Y Park, YC Ohnishi, N Sugiyama, T Kurita, T Kissei Pharmaceut Co Ltd Div Discovery Res Nagano 3998304 Japan Kinki Univ Sch Med Dept Urol Osaka 589 Japan

The objective of the present study was to evaluate the effects of a selective beta(3)-adrenoceptor agonist, (R,R)-5-[2-[[2-(3-chlorophenyl)- 2- hydroxyethyl]- amino] propyl]-1,3-benzodioxole-2,2-dicarboxylate (CL 316243), on the acutely obstructed ureter in anesthetized dogs. After a complete ureteral obstruction produced by the inflation of a balloon catheter placed within the left lower ureter, the intraluminal ureteral pressure gradually rose to reach a plateau of similar to 52.5 mm Hg. Intravenous administration of isoproterenol (a nonselective beta-adrenoceptor agonist;10 mu g/kg) and CL 316243 (1 mu g/kg) significantly decreased this elevated ureteral pressure (by 74.1 and 77.2%, respectively), with the reduction more sustained with CL 316243 than with isoproterenol. In addition, under both isoproterenol and CL 316243, urine flow (which had been interrupted by the balloon) was resumed, resulting in further sustained decreases in ureteral pressure. The mean blood pressure decreased and heart rate increased after the administration of both drugs, but these changes were greater in the isoproterenol group than in the CL 316243 group. In contrast, i.v. administration of butylscopolamine (an anticholinergic agent;1000 mu g/kg) had no evident effects on ureteral pressure or on urine flow. The increase in left kidney weight seen after ureteral obstruction was suppressed by CL 316243. We conclude that the selective beta(3)-adrenoceptor agonist tested appears to be more useful than isoproterenol for reducing ureteral pressure above the obstructed site and for promoting ureteral relaxation and increasing urine flow around the point of obstruction in dogs.

关键词：肾上腺素能β激动剂/药理学麻醉血压/药物作用丁溴东莨菪碱/药理学导管插入术二恶茂类/药理学心率/药物作用异丙肾上腺素/药理学肾小管/药物作用肾小管/病理学毒蕈碱拮抗剂/药理学时间因素输尿管梗阻/病理生理学尿动力学动物狗女(雌)性男(雄)性

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HOST-MEDIATED ASSAY WITH YEAST AND RATS USING PROBENECID (BENEMID) TO BLOCK RENAL TUBULAR EXCRETION OF CYCLOPHOSPHAMIDE METABOLITES

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MUTATION RESEARCH 1975年第1期28卷 57-61页

作者： SIEBERT, D UNIV FREIBURG INST FOREST BOTFREIBURGFED REP GER

The genetic activity of cyclophosphamide (Cy) was tested in the host-mediated assay (injection of yeasts into the peritoneal cavity of rats) modified by the use of probenecid (Pro) (Benemid®) to block the renal tubular excretion of the genetically active metabolites. The genetic test system used was the induction of mitotic gene conversion in two unlinked loci of a diploid strain of Saccharomyces cerevisiae . By this method the genetic effect of Cy was doubled in comparison with the case of administering Cy alone. Compared with the animals which received only Pro, increases of conversion frequencies of 20 times in the ade2 locus and of 15 times in the trp5 locus were found.

关键词：染色体图环磷酰胺/代谢环磷酰胺/药理学肾小管/药物作用肾小管/生理学有丝分裂诱变剂/药理学腹膜腔丙磺舒/药理学重组遗传酿酒酵母菌/药物作用动物男(雄)性大鼠

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Attenuation of cisplatin-induced acute renal failure is associated with less apoptotic cell death

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JOURNAL OF LABORATORY AND CLINICAL MEDICINE 1999年第6期134卷 649-658页

作者： Zhou, H Miyaji, T Kato, A Fujigaki, Y Sano, K Hishida, A Hamamatsu Univ Sch Med Dept Med Hamamatsu Shizuoka 4313192 Japan

To clarify the pathophysiologic role of apoptosis in acute renal failure (ARF), we examined whether the attenuation of cisplatin-induced ARF is associated with the change in the degree of apoptotic cell death. The administration of cisplatin (CDDP) (6 mg/kg body weight) in rats induced ARF at day 5, as manifested by a significant increase in serum creatinine (Scr) and tubular damage. CDDP-induced apoptotic cell death was confirmed by electron microscopic examination, agarose gel electrophoresis, and increased cells positive for TaT-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) in the outer medulla of the kidney. Treatment with dimethylthiourea (DMTU)-a scavenger of hydroxyl radicals--or glycine abrogated CDDP-induced increases in Scr, the tubular damage score, and the number of TUNEL-positive cells. Pretreatment with uranyl acetate (UA) induced a significant expression of Bcl-2 in the kidney and ameliorated CDDP-induced increases in Scr, the tubular damage score, and TUNEL-positive cells in the outer stripe of the outer medulla, Our findings indicate (1) that the attenuation of CDDP-induced ARF was associated with less apoptotic cell death and (2) that the induction of the anti-apoptotic protein Bcl-2 attenuated apoptosis and tubular damage. Our results suggest that apoptotic cell death may play an important role in the development of cisplatin-induced ARF.

关键词：肾髓质/药物作用肾髓质/代谢肾髓质/病理学肾小管/药物作用肾小管/代谢肾小管/超微结构有机金属化合物/药理学原癌基因蛋白质c-bcl-2/生物合成大鼠 Sprague-Dawley 硫脲/类似物和衍生物硫脲/治疗应用男(雄)性大鼠

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Studies on the mechanisms and kinetics of apoptosis induced by microinjection of cytochrome c in rat kidney tubule epithelial cells (NRK-52E)

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AMERICAN JOURNAL OF PATHOLOGY 2000年第2期156卷 637-649页

作者： Chang, SH Phelps, PC Berezesky, IK Ebersberger, ML Trump, BF Univ Maryland Sch Med Dept Pathol Baltimore MD 21201 USA

Recent reports substantiating the role of cytochrome c in the induction of apoptosis led us to examine the kinetics and mechanisms involved in this process as an extension of our ongoing studies of cell injury and cell death. Microinjection of cytochrome c into NRK-52E kidney cells produced rapid apoptosis, which usually began within 30 minutes and reached a maximum of 60-70% by 3 hours. The changes that occurred included four phases: an initial shrinkage phase, an active phase, a spherical phase, and a necrotic phase. For morphological purposes, the progressive changes were followed by phase-contrast and fluorescence microscopy, transmission and scanning electron microscopy, and time-lapse video microscopy, Cells first showed shrinkage, then displayed multiple pseudopods, which rapidly extended and retracted, giving the cells a bosselated appearance. During this active phase there was chromatin condensation, mitochondria were swollen but retained membrane potential, and the endoplasmic reticulum was dilated. Within 2-4 hours, active-phase cells became spherical and smooth-surfaced but were still alive, the nuclei showed chromatin clumping, the mitochondria underwent high-amplitude swelling but retained membrane potential, the endoplasmic reticulum was highly dilated, acid many large apical vacuoles were present. Elevation of [Ca2+](i) was seen at the late spherical phase, shortly before cell death. Pretreatment with the caspase 3 inhibitor (Ac-DEVD-CHO) prevented apoptosis, whereas overexpression of Bcl-2 did not. Depletion of cellular ATP by cyanide inhibition of energy metabolism prevented cytochrome c from inducing the active and later phases of apoptosis, The results clearly indicate that cytochrome c-induced apoptosis is a dynamic and energy-requiring process that has a distinct active and spherical phase before cell death.

关键词：腺苷三磷酸/代谢细胞凋亡/生理学钙/代谢半胱氨酸天冬氨酸蛋白酶3 半胱氨酸天冬氨酸蛋白酶/生理学细胞系细胞色素c组/药理学胞质溶胶/代谢上皮细胞/药物作用上皮细胞/生理学上皮细胞/超微结构肾小管/细胞学肾小管/药物作用肾小管/生理学动力学微量注射显微镜检查电子显微镜检查电子扫描原癌基因蛋白质c-bcl-2/代谢动物大鼠

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Cyclosporin exerts a direct fibrogenic effect on human tubulointerstitial cells:: Roles of insulin-like growth factor I, transforming growth factor β₁, and platelet-derived growth factor

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第1期289卷 535-542页

作者： Johnson, DW Saunders, HJ Johnson, FJ Huq, SO Field, MJ Pollock, CA Royal N Shore Hosp Univ Sydney Dept Med Sydney NSW 2065 Australia

To assess the direct fibrogenic effects of cyclosporin A (CyA) on the human tubulointerstitium, primary cultures of human renal proximal tubule cells (PTC) and renal cortical fibroblasts (CF) were incubated for 24 h with various concentrations of CyA. Cytotoxicity was confirmed in both cell populations by dose-dependent inhibition of thymidine incorporation, viability, and PTC apical sodium-hydrogen exchange activity (ethylisopropylamiloride-sensitive apical Na-22(+) uptake). Compared with controls, both 500 and 1000 ng/ml CyA significantly stimulated CF collagen synthesis (proline incorporation 4.6 +/- 0.4, 6.5 +/- 0.8, and 7.1 +/- 1.0%, respectively;p <.05) and inhibited matrix metalloproteinase-2 (100%, 85.7 +/- 10.0%, and 38.8 +/- 9.2%) and matrix metalloproteinase-9 activity (100%, 110.6 +/- 19.0%, and 49.9 +/- 12.8%), CyA did not affect CF secretion of transforming growth factor beta(1), but markedly stimulated insulin-like growth factor-I (IGF-I) secretion and inhibited secretion of both IGF-I binding protein-(IGFBP)-3 and IGFBP-2, CyA-induced CF collagen synthesis was abrogated by 5 mu g/ml anti-IGF-I receptor antibody, but not by 5 mu g/ml murine nonimmune globulin. Increasing concentrations of CyA progressively augmented PTC secretion of the fibrogenic cytokines transforming growth factor-beta(1) and platelet-derived growth factor. These results indicate that clinically relevant concentrations of CyA are directly toxic to PTC and CF, irrespective of hemodynamic effects, and promote interstitial fibrosis by inhibiting matrix degradation and stimulating cortical fibroblast collagen synthesis via induction of autocrine IGF-I action. The latter effect may be further accentuated by the ability of CyA to augment secretion of transforming growth factor beta(1) and platelet-derived growth factor by PTCs.

关键词：细胞存活/药物作用细胞培养的胶原/生物合成培养基条件性环孢菌素/毒性纤维化明胶/代谢免疫抑制剂/毒性胰岛素样生长因子Ⅰ/生理学肾皮质/药物作用肾皮质/代谢肾皮质/病理学肾小管/药物作用肾小管/代谢肾小管/病理学肾小管近端/药物作用肾小管近端/病理学金属内肽酶类/代谢血小板源性生长因子/生理学转化生长因子β/生理学女(雌)性人类男(雄)性中年人

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Nephroprotective effect of treatment with calcium channel blockers in spontaneously hypertensive rats

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 2000年第3期294卷 948-954页

作者： Sabbatini, M Vitaioli, L Baldoni, E Amenta, F Univ Camerino Dipartimento Sci Farmacol & Med Sperimentale Sez Anat Umana I-62032 Camerino Italy Univ Camerino Dipartimento Sci Morfol & Biochim Comparate I-62032 Camerino Italy

The influence of hypertension and of treatment with some dihydropyridine-type Ca2+ channel blockers and with the nondihydropyridine-type vasodilator hydralazine on the morphology of kidney was investigated in 26-week-old spontaneously hypertensive rats (SHR) and in age-matched Wistar-Kyoto rats. Fourteen-week-old SHR were treated for 12 weeks with a nonhypotensive dose of lercanidipine or with equihypotensive doses of lercanidipine, manidipine, nicardipine, and hydralazine. In control SHR, systolic pressure values were significantly higher in comparison with Wistar-Kyoto rats. Treatment with the low dose of lercanidipine did not reduce systolic blood pressure in SHR, whereas the higher dose of lercanidipine or other compounds tested significantly decreased systolic pressure values. Glomerular hypertrophy accompanied by signs of glomerulosclerosis, increase of mesangial cells, and convoluted tubules degeneration were observed in control SHR. Hypotensive doses of Ca2+ antagonists countered glomerular injury, the increase of mesangial cells, the reduction of capsular space, and tubular degeneration. Hydralazine, in spite of its hypotensive activity, displayed a slight nephroprotective action. The nonhypotensive dose of lercanidipine countered in part glomerular injury, narrowing of capsular space, and tubular degeneration, and decreased mesangial cell augmentation in SHR. These results suggest that treatment with dihydropyridine-type Ca+2 antagonists counters hypertensive glomerular and tubular changes occurring in SHR. The demonstration of nephroprotection by the nonhypotensive dose of lercanidipine suggests that the renal effects of the compound may be in part unrelated to its hemodynamic activity.

关键词：血压/药物作用钙通道阻滞药/药理学二氢吡啶类/药理学肼苯哒嗪/药理学肾/血液供给肾/药物作用肾/病理学肾皮质/药物作用肾皮质/病理学肾小球/药物作用肾小球/病理学肾小管/药物作用肾小管/病理学尼卡地平/药理学大鼠近交SHR 大鼠近交WKY 血管舒张药/药理学动物男(雄)性大鼠

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Effects of hemoglobin-based oxygen-carrying solutions in anesthetized rats with acute ischemic renal failure

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JOURNAL OF LABORATORY AND CLINICAL MEDICINE 2000年第1期135卷 73-81页

作者： Lieberthal, W Fuhro, R Andry, C Valeri, CR Boston Univ Med Ctr Div Renal Boston MA 02118 USA Boston Univ Med Ctr Div Pathol Boston MA 02118 USA Boston Univ Med Ctr Naval Blood Res Lab Boston MA 02118 USA

The effects of three hemoglobin solutions were compared with those of iso-oncotic human serum albumin in rats with ischemic renal failure and sham-operated controls. Unmodified and alpha-alpha cross-linked hemoglobins both increase mean arterial pressure and systemic vascular resistance and reduce cardiac output substantially and to a comparable extent. In contrast, o-raffinose cross-linked hemoglobin has no deleterious effect on any of these parameters, In sham-operated rats unmodified hemoglobin reduces the glomerular filtration rate (GFR) by approximately 30%, whereas neither of the two cross-linked hemoglobins has any adverse effect on GFR in this group. None of the three hemoglobin solutions exacerbated the degree to which GFR was reduced by ischemia-reperfusion injury. Also, the degree of tubular necrosis induced by ischemia-reperfusion injury was also comparable in all groups. We conclude the following: (1) o-raffinose cross-linking, but not alpha-alpha cross-linking, ameliorates the effects of unmodified hemoglobin on vascular resistance and cardiac output;(2) both forms of cross-linking reduce the nephrotoxicity exhibited by unmodified hemoglobin in sham-operated rats;and (3) none of the hemoglobin solutions exacerbate renal injury induced by ischemia-reperfusion.

关键词：阿司匹林/类似物和衍生物阿司匹林/药理学血压/药物作用肾小球滤过率/药物作用心率/药物作用血红蛋白类/药理学肾功能衰竭急性/代谢肾功能衰竭急性/病理学肾功能衰竭急性/病理生理学肾小管/药物作用肾小管/病理学肾小管/病理生理学坏死氧/代谢棉子糖/类似物和衍生物棉子糖/药理学大鼠 Sprague-Dawley 再灌注损伤/代谢再灌注损伤/病理学再灌注损伤/病理生理学血清白蛋白/药理学钠/尿血管收缩/药物作用动物人类男(雄)性大鼠

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An evaluation of the carcinogenic hazard of 1,4-dychlorobenzene based on internationally recognized criteria

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REGULATORY TOXICOLOGY AND PHARMACOLOGY 1999年第1期29卷 64-79页

作者： Barter, JA Sherman, JH PPG Ind Inc Pittsburgh PA 15272 USA Solutia Inc St Louis MO 63166 USA

1,4-Dichlorobenzene (1,4-DCB) was shown to induce the formation of male rat renal tubule tumors and male and female mouse liver tumors when administered in a chronic bioassay. Since the original carcinogenicity findings, an extensive body of mechanistic information has been developed to elucidate the mode of action by which 1,4-DCB induces these effects and to evaluate the human relevance of the observed animal tumors. In addition, some regulatory and authoritative bodies (U.S. EPA and IARC) have developed rigorous scientific criteria for the amount and types of evidence needed to establish that a material causes kidney toxicity and tumors in male rats through a specific mechanism, alpha-2u-globulin nephropathy. This paper summarizes the mechanistic data developed for 1,4-DCB, which affords an understanding of the lack of human relevance of the male rat renal tubule tumors and mouse Liver tumors;assesses that mechanistic data set utilizing the defined set of evaluation criteria formulated by U.S. EPA and IARC for alpha-2u-globulin nephropathy;and discusses the predictive power of mechanistic data developed to elucidate the mode of action of 1,4-DCB in inducing mouse liver tumors. Finally, there is a discussion of how some, but not all, regulatory and authoritative bodies have incorporated this substantial mechanistic data set for 1,4-DCB into their cancer hazard evaluations and concluded that 1,4-DCB presents Little, if any, cancer hazard to humans. (C) 1999 Academic Press.

关键词：致癌物/毒性氯苯类/毒性杀虫药/毒性肾肿瘤/化学诱导肾小管/药物作用肝肿瘤实验性/化学诱导小鼠近交C3H 小鼠近交C57BL 大鼠近交F344 动物女(雌)性人类男(雄)性小鼠大鼠

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