纳洛酮对大鼠肾脏缺血再灌注损伤的保护作用

纳洛酮对大鼠肾脏缺血再灌注损伤的保护作用孙成春贾暖张霞１郝俊文２王景祥（济南军区总医院药理科、１病理科、２泌尿外科，济南２５００３１）１９９６－１０－２３收稿，１９９７－０３－０７修回作者简介：孙成春，女，３２岁，硕士，主管药师；王景祥，男，６２岁，...     

人参皂甙Rb1对大鼠急性肾小管损伤的保护作用

目的:观察单体人参皂甙R b1(R b1)对缺血/再灌注(I/R)所致急性肾小管损伤的保护作用,并探讨其机制。方法:夹闭大鼠双肾动脉造成急性I/R肾损伤模型,光镜观察形态学改变,应用图像分析系统计算肾小管坏死面积,观察血清肌酐(Scr)变化,测定血清丙二醛(M DA)及超氧化物歧化酶(SOD)。结果:与对照组相比,R b1组肾小管坏死面积明显减少[(32.32%±18.65%)vs(56.87%±27.94%)]P<0.01;肾功能损害减轻;I/R 48 h,R b1组小管上皮增生明显活跃;血清M DA升高及SOD下降的程度减轻。结论:R b1能够保护肾I/R肾小管损伤,加速小管上皮细胞的修复,抗氧化作用参与了其保护机制。     

百里醌对大鼠肾脏缺血再灌注损伤的保护作用

目的 观察百里醌对大鼠肾缺血再灌注损伤（ischemia-reperfusion injury,IRI）的保护作用。方法 60只健康SD大鼠,♂,随机分成正常组、模型组和百里醌组。模型组和百里醌组大鼠建立肾脏缺血再灌注模型,百里醌组在再灌注同时腹腔注射百里醌（40 mg·kg^-1）。HE染色观察肾脏组织病理改变;测定血清指标肌酐（Scr）、尿素氮（BUN）以及肾组织指标丙二醛（MDA）、超氧化物岐化酶（SOD）;采用原位缺口末端标记检测肾小管上皮细胞凋亡指数（AI）;RT-PCR检测肾脏组织中的caspase-3,Bax和TNF-α和IL-1β的表达情况。结果 模型组可见肾小管扩张,肾小管上皮细胞坏死等,经百里醌治疗后明显改善。与模型组相比,百里醌组凋亡指数明显降低（P<0.05）。与正常组比较,模型组SOD活性显著降低（P<0.05）,MDA含量、Scr和BUN显著升高（P<0.05）;与模型组比较,百里醌组SOD活性显著升高（P<0.05）,MDA含量、Scr和BUN显著降低（P<0.05）;与正常组比较,模型组中caspase-3、Bax、TNF-α和IL-1β mRNA表达水平明显增高（P<0.05）。和模型组比较,百里醌组中caspase-3、Bax、TNF-α和IL-1β表达明显减低（P<0.05）。结论 百里醌可减轻氧化应激水平,发挥抗炎及抗凋亡作用,从而在肾脏IRI过程中发挥保护作用。     

慢性染砷对大鼠肾脏DNA损伤的影响

目的本实验通过用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)实验方法探讨慢性染砷对大鼠肾脏组织细胞DNA的影响及其可能机制。方法60只3周龄Wistar大鼠随机分为低剂量染砷组(A1)、高剂量染砷组(A2)和对照组(C),分别自由饮用10.0mg/L、40.0mg/L的三氧化二砷水溶液和自来水,喂养6个月后,处死大鼠,取新鲜的肾脏组织进行SCGE实验。结果染砷组肾脏组织细胞DNA损伤明显高于对照组,A2、A1组与对照组比较,P<0.01。结论慢性染砷可引起大鼠肾脏组织细胞DNA的损伤。     

丹参对大鼠肾脏缺血再灌注损伤的保护作用机制的研究

目的：探讨丹参对大鼠肾脏缺血再灌注损伤的保护作用。方法：雄性Wistar大鼠随机分为4组,假手术组,模型组（肾脏缺血再灌注损伤）,治疗1组（肾脏缺血再灌注损伤前24h给予药物）,治疗2组（肾脏缺血再灌注损伤后12h给予药物）。双侧肾动脉夹闭22min,制作动物模型;比色法测定血清肌酐和血清尿素氮;Westernblotting检测肾脏组织中,天冬氨酸特异性半胱氨酸蛋白酶（caspase-3）的蛋白表达;TUNEL法检测肾脏上皮细胞凋亡。结果：模型组与假手术组比较,大鼠肾脏功能明显减退（P〈0.05）;肾脏组织中,caspase-3蛋白质表达显著增加（P〈0.05）,大量肾小管上皮细胞凋亡（P〈0.05）。再灌注前24h给予药物,能够显著改善肾脏功能（P〈0.05）,并且显著下调caspase-3的蛋白表达（P〈0.05）,减轻肾脏上皮细胞凋亡（P〈0.05）,再灌注后12h给药,不能改善肾脏功能,也不能显著下调caspase-3的蛋白表达（P〉0.05）。结论：再灌注前给予丹参,能够抑制肾脏缺血再灌注损伤诱导的肾脏上皮细胞凋亡,对肾脏缺血再灌注损伤具有保护作用。     

大黄和泽泻提取物对二甘醇致小鼠肾脏损伤的保护作用研究

目的:研究大黄和泽泻提取物对二甘醇(DEG)致小鼠肾脏损伤的保护作用及其机制。方法:采用ig给予DEG方法建立小鼠肾脏损伤模型,ig给予大黄和泽泻提取物后测定血清中肌酐(Cr)和尿素氮(BUN)浓度,测定超氧化物歧化酶(SOD)和谷胱甘肽过氧化酶(GSH-PX)活性及丙二醛(MDA)含量变化。结果:小鼠ig给予DEG后有明显中毒表现,肾脏体质量比增加,血清中Cr和BUN水平升高,肾组织SOD和GSH-PX活性降低、MDA含量增加;给予大黄和泽泻提取物后小鼠中毒表现明显减轻,肾脏体质量比减少,血清中Cr和BUN水平明显降低,肾组织SOD和GSH-PX活性升高、MDA含量降低。结论:大黄和泽泻提取物对DEG所致小鼠肾脏损伤具有明显的保护作用,该作用与其改善肾脏抗氧化酶活性及抑制脂质过氧化反应有关。     

丙二醇质量控制方法和二甘醇检查方法探讨

目的对丙二醇质量控制方法和二甘醇检查方法进行理论探讨和实践研究。方法在对丙二醇和二甘醇进行简要介绍的基础上,通过分析《中国药典（2005年版,二部）》对丙二醇质量控制方法,指出目前药典方法存在的不足,并通过实验研究提出解决方案。结果药典规定的质量控制方法不能完全保证丙二醇的质量,新的质量控制方法应包括丙二醇的含量测定和二甘醇检查。结论GC—MS法可作为丙二醇含量测定和二甘醇检查的参考方法。     

GC法测定葛根素注射液中丙二醇和二甘醇的残留量

采用气相色谱法测定葛根素注射液中丙二醇和二甘醇的残留量.用DB-WAXetr毛细管柱,检测器温度为280℃.丙二醇的测定在49.69～993.8цg·mL-1范围内线性关系良好,平均回收率为98.4%,RSD=1.2%(n=9);二甘醇在11.09～110.9цg·mL-1范围内线性关系良好,平均回收率为99.2%,RSD=1.4%(n=9).方法简单、灵敏度高.     

胶束电动色谱法测定硫芥水解产物硫二甘醇

目的　硫芥是一种化学性质非常活泼的糜烂性毒剂 ,其水解反应的主要产物是硫二甘醇 (TDG)。本实验采用胶束电动色谱法 (MEKC)对微量TDG进行检测。方法　采用P/ACE5 0 0 0毛细管电泳仪 ,有效分离长度为 5 0cm、内径 5 0 μm的石英毛细管柱 ,UV检测器 (检测波长 2 0 0nm) ;温度 2 5℃ ;工作电压2 0kV ;电迁移进样。pH为 9.2的 0 .0 1mol·L- 1硼砂缓冲盐体系 (其中SDS浓度为 5 0mmol·L- 1) ,对不同浓度的TDG溶液进行检测。并对其中工作缓冲液的pH值、测定波长及进样方式等实验条件进行了优化。结果　在优化的工作条件下 ,TDG浓度在10～ 5 0mg·L- 1范围内 ,其浓度与峰高有良好的线性关系 ,相关系数r =0 .972 3。方法的回收率为10 0 .6 %。结论　用MEKC对水中TDG进行检测 ,方法简便 ,灵敏度高 ,检测时间短 ,运行成本低 ,有较高的应用价值。     

番茄红素对肾脏缺血再灌注损伤大鼠的保护作用

目的 探究番茄红素(LP)通过抑制硫氧还蛋白相互作用蛋白/NOD样受体热蛋白结构域相关蛋白3(TXNIP∕NLRP3)信号通路保护大鼠肾脏缺血再灌注损伤(IRI).方法 100只大鼠随机分为对照组(n=25)、模型组(n=25)及低剂量实验组(n=25)、高剂量实验组(n=25).模型组及低/高剂量实验组均采用右侧肾切...     

气相色谱法测定聚山梨酯80中二甘醇的含量

目的建立一种气相色谱法测定聚山梨酯80中二甘醇的含量.方法色谱柱:以聚乙二醇(PEG-20M)为固定液的极性毛细管柱(30m×0.32mm,0.50μm);检测器:氢火焰离子检测器(FID);检测器温度230℃;进样口温度200℃;柱温度:程序升温,初始温度60℃,保持3min,再以10℃·min-1升温至230℃保持10min;载气:高纯氮气.结果二甘醇线性范围为0.025～0.25mg·mL-1(r=0.9993);平均回收率101.57%(RSD=1.505%,n=5).结论本方法简单、结果准确、重现性好,可用于聚山梨酯80中二甘醇含量的测定.     

Screening method for ethylene glycol and diethylene glycol in glycerin-containing products

This paper describes a capillary gas chromatographic method with flame ionization detection for the identification/quantification of ethylene glycol (EG) and diethylene glycol (DEG) in glycerin. The validation study shows that the proposed method is specific, sensitive, precise, and accurate. The linear range of the method was 0.013–0.031 mg/mL for EG and 0.012–0.030 mg/mL for DEG. Wider ranges may be achievable but were not investigated. The limit of detection of EG and DEG were determined as 0.0018% and 0.0036% (w/w) respectively, and at this concentration the signal-to-noise ratios for EG and DEG were approximately 3:1. The method was also used to determine EG and DEG in toothpaste. The results were compared to those obtained by thin-layer chromatography (TLC) and showed greater sensitivity and specificity.     

Toxicokinetics of diethylene glycol (DEG) in the rat

Oral doses of 1 and 5 ml/kg¹⁴C-diethylene glycol (DEG) given to rats were rapidly and almost completely absorbed, the invasion constants being 2.95 h^– and 4.24 h^–1. The kinetics of invasion were determined with the method of residuals (Rowland and Tozer 1989) and by reconstruction of the invasion curves according to Kübier (1970).¹⁴C-DEG was rapidly distributed from the blood into the organs and tissues in the order kidneys > brain > spleen > liver > muscle > fat, i.e. the same order as the blood flow. The relative volume of distribution, app. V_D, was determined at 298 ml, indicating distribution over the whole body. After oral doses of 1, 5, and 10 ml¹⁴C-DEG/kg 64, 87, and 91% of¹⁴C activity in rat blood disappeared in 12–16 h with a half-life of 3.4 h and the remaining 9, 5, and 4% with half-lives of 39 h, 45 h, and 49 h. A total of 73–96% of¹⁴C activity in blood was excreted with the urine and 0.7–2.2% with the faeces. From the cumulative urinary excretion kinetics half-lives of 6 h were determined for doses of 1 and 5 ml/kg and 10 h for the dose of 10 ml/kg. After doses of 5 ml/kg and 10 ml/kg¹⁴C-DEG semi-logarithmic plots of elimination rate versus time were constant for 5 and 9 h, respectively, indicating that DEG accelerated its renal elimination by inducing osmotic diuresis. Thereafter urinary excretion followed first order kinetics with elimination half-lives of 3.6 h. After oral doses of 5 ml/kg¹⁴C-DEG given to rats of 336 g body weight with an app. V_D of 297 ml, the total clearance of¹⁴C activity was determined at 63 ml/h, and the renal clearance of unmetabolized DEG was 66 ml/h. The ratio of Cl_DEG to Cl_inulin = 0.64 indicated that DEG and its metabolite 2-hydroxyethoxyacetate (2-HEAA) were reabsorbed from the tubuli into the blood capillaries. DEG produced metabolic acidosis, which was completely balanced after doses of 1 and 5 ml/kg, but doses greater than 10 ml/kg produced non-compensated metabolic acidosis, hydropic degeneration of the tubuli, oliguria, anuria, accumulation of urea-N, and death in uraemic coma.Second communication on the effects of DEG in the rat; first communication, Lenk et al. (1989)     

Identification and quantification of ethylene glycol and diethylene glycol in plasma using gas chromatography-mass spectrometry

A method for the gas chromatographic-mass spectrometric identification and quantification of ethylene glycol and diethylene glycol in plasma is described. Such a method is necessary in clinical and forensic toxicology to diagnose probable intoxication and to control the efficacy of detoxification. For sample preparation, the glycols were isolated using acetone after the addition of 1,3-propylene glycol as internal standard. The glycols were then esterified by pivalic acid (pivalic acid anhydride, triethylamine and methanol, 70° C, 15 min) to improve their gas chromatographic characteristics. The glycols were first identified by a comparison of the full mass spectra with reference spectra and then quantified. Therefore, the peak area ratio in the total ion chromatogram (ethylene glycol or diethylene glycol/1,3-propanediol) of the sample was compared with the calibration curve in which the peak area ratios of the standards (0.05, 0.1, 0.5, 1 and 2 g/l), prepared in the same way, were plotted versus their concentrations. The method was linear at least from 0.05 to 2 g/l, with a detection limit of <0.01 g/l. The analytical recoveries were 99.2–102.9% for the different concentrations. Precision studies show coefficients of variation of 3.0–6.3% for the different concentrations.Some of these results were reported at the Spring meeting of the German Pharmacological Society in Mainz, FRG, March 11–14, 1986 (Maurer et al. 1986). They are part of the M. D. thesis of C. Kessler (Kessler 1988).     

Detection of diethylene glycol adulteration in propylene glycol--method validation through a multi-instrument collaborative study

Four portable NIR instruments from the same manufacturer that were nominally identical were programmed with a PLS model for the detection of diethylene glycol (DEG) contamination in propylene glycol (PG)-water mixtures. The model was developed on one spectrometer and used on other units after a calibration transfer procedure that used piecewise direct standardization. Although quantitative results were produced, in practice the instrument interface was programmed to report in Pass/Fail mode. The Pass/Fail determinations were made within 10s and were based on a threshold that passed a blank sample with 95% confidence. The detection limit was then established as the concentration at which a sample would fail with 95% confidence. For a 1% DEG threshold one false negative (Type II) and eight false positive (Type I) errors were found in over 500 samples measured. A representative test set produced standard errors of less than 2%. Since the range of diethylene glycol for economically motivated adulteration (EMA) is expected to be above 1%, the sensitivity of field calibrated portable NIR instruments is sufficient to rapidly screen out potentially problematic materials. Following method development, the instruments were shipped to different sites around the country for a collaborative study with a fixed protocol to be carried out by different analysts. NIR spectra of replicate sets of calibration transfer, system suitability and test samples were all processed with the same chemometric model on multiple instruments to determine the overall analytical precision of the method. The combined results collected for all participants were statistically analyzed to determine a limit of detection (2.0% DEG) and limit of quantitation (6.5%) that can be expected for a method distributed to multiple field laboratories.     

Pivotal role of pericytes in kidney fibrosis

1. Kidney pericytes were recently identified as collagen Iα1-producing cells in healthy kidney, but the developmental, physiological and pathological roles of kidney pericytes remain poorly understood. Pericytes are stromal-derived cells that envelop and have intimate connections with adjacent capillary endothelial cells (EC). Recent studies in the eye and brain have revealed that pericytes are crucial for angiogenesis, vascular stability and vessel integrity. 2. In response to kidney injury, pericytes promptly migrate away from the capillary wall into the interstitial space. Here, pericytes are activated and differentiate into scar-forming myofibroblasts. In the absence of pericytes, peritubular capillaries are destabilized, leading to vascular regression. Consequently, capillary loss and fibrosis following kidney injury are intimately linked and hinge centrally around pericyte detachment from EC. 3. Kinetic mathematical modelling has demonstrated that pericytes are the major source of myofibroblasts in the fibrotic kidney. Comprehensive genetic fate mapping studies of nephron epithelia or kidney stroma has demonstrated that epithelial cells do not migrate outside of the epithelial compartment to become myofibroblasts; rather, interstitial pericytes are progenitors of scar-forming myofibroblasts. Bidirectional signalling between pericytes and EC is necessary for pericyte detachment from peritubular capillaries. 4. In the present review, we summarize the pathologically vital roles of kidney pericytes in fibrosis, including our new findings. The study of kidney pericytes and endothelial-pericyte cross-talk will identify novel therapeutic targets for currently incurable chronic kidney diseases.     

前列地尔治疗急性肾损伤的临床疗效

目的探讨前列地尔治疗急性肾损伤(AKI)的临床疗效。方法AKI病人70例,随机分成治疗组(n=38)和对照组(n=32)。对照组给予常规治疗。治疗组在常规治疗的基础上再给予前列地尔每日剂量80μg,静脉滴注,连用10～14d。结果治疗组少尿持续时间为(8±s 4)d,对照组为(12±5)d,2组比较差异有显著意义(P<0.05)。对照组21例(66%)行血液透析治疗,平均透析次数(6.8±2.6)次;治疗组14例(37%)行血液透析治疗,平均透析次数(4.1±2.3)次,2组比较差异显著(P<0.05)。治疗组死亡9例(24%),对照组死亡15例(47%),差异有显著意义(P<0.05)。结论前列地尔明显缩短AKI疗程、改善病人预后,早期诊治可以改善AKI的预后。     

埃他卡林对脂多糖、油酸、二甘醇等所致肾脏损伤的影响

目的观察埃他卡林对脂多糖、油酸和二甘醇等不同因素所致肾脏损伤的影响。方法采用大鼠股静脉注射2%醋酸铅致敏后注射脂多糖1μg（1ml/kg）,4h造成内毒素休克肾损伤模型,大鼠左肾动脉注射油酸0.15ml/kg,24h造成油酸所致肾损伤模型,分别在造模前3d及1h,以埃他卡林1,3,9mg/（kg.d）灌胃给药;小鼠腹腔注射二甘醇10g/kg后,立即以埃他卡林1,3,9mg/（kg.d）灌胃给药6d,第7天造模成功。3种模型建立后,观察血清肌酐、尿素氮水平和肾脏形态学变化,评价肾脏功能。结果（1）内毒素性休克大鼠血清肌酐和尿素氮水平显著升高,组织病理显示有肾小球微血栓、肾小管上皮肿胀和管腔内蛋白管型形成。埃他卡林9mg/kg组能明显降低血清尿素氮和肌酐水平,改善上述病理变化。（2）左肾动脉注射油酸大鼠血清肌酐和尿素氮水平显著升高,组织病理显示肾小球内皮细胞坏死,球囊腔减小,肾小管间质充血且管腔内有蛋白管型形成。埃他卡林对油酸所致大鼠肾脏损伤无显著性改善作用。（3）二甘醇肾损伤小鼠血清肌酐水平显著性升高,埃他卡林9mg/kg组血清肌酐水平恢复至正常。结论埃他卡林不适合于脂多糖、油酸所致肾脏损伤的防治,埃他卡林可否用于二甘醇所致肾损伤的治疗值得进一步研究。     

P38MAPK 抑制剂 SB203580对高糖诱导HK-2细胞转分化的影响

摘要： 目的 探讨不同浓度 P38 丝裂原活化蛋白激酶 （P38MAPK） 抑制剂 SB203580 在高糖诱导肾小管上皮细 胞-肌成纤维细胞转分化 （TEMT） 过程中的机制及其较佳作用浓度。方法 体外培养人近端肾小管上皮细胞 （HK- 2） 并分为对照组 （5.5 mmol/L GS）、 DMSO 组 （5.5 mmol/L GS + 30 μmol/L SB203580 等体积的 DMSO）、 高糖组 （30 mmol/L GS）, 以及 30 mmol/L GS +5、 10、 20、 30 μmol/LSB203580 处理的 S5、 S10、 S20 及 S30 组, 干预 48 h。四甲基偶 氮唑蓝 （MTT） 法检测细胞增殖情况, 计算半数抑制浓度 （IC50 ）; 选取对照组、 高糖组、 S30 组, Western blot 法检测 P38MAPK、 P-P38MAPK 及α-平滑肌肌动蛋白 （SMA） 的表达、 免疫荧光法检测α-SMA 的表达。结果 （1） 与对照组 相比, DMSO 对 HK-2 细胞增殖无显著抑制作用 （P > 0.05）; 高糖组、 S5 组 HK-2 细胞增殖增多 （P < 0.05）; S20、 S30 组 HK-2 细胞增殖减少 （P < 0.05）。与高糖组相比, S5、 S10、 S20、 S30 组细胞增殖均受到抑制 （P < 0.05）。（2） 与对照 组相比, 高糖组、 S30 组 P-P38MAPK 表达量增高 （P < 0.05）。与高糖组相比, S30 组 P-P38MAPK 的表达量降低 （P < 0.05）。3 组 P38MAPK 表达量无显著差异 （P > 0.05）。（3） 与对照组相比, 高糖组、 S30 组α-SMA 表达量增高 （P < 0.05）。与高糖组相比, S30 组α-SMA 表达量降低 （P < 0.05）。结论 30 mmol/L GS 可以诱导 HK-2 细胞 TEMT;30 μmol/L SB203580 是抑制 HK-2 细胞 TEMT 的较佳抑制浓度, SB203580 可能通过下调 P-P38MAPK 表达, 从而抑制 HK-2细胞增殖及胞浆中α-SMA 的表达, 延缓 TEMT 进程。     

醛固酮对人近端肾小管上皮细胞结缔组织生长因子mRNA表达的影响

目的探讨醛固酮(ALD)在体外是否能够促进人近端肾小管上皮细胞株(HK2)结缔组织生长因子(CTGF)的表达.方法采用体外培养的HK2细胞,分别观察不同浓度(10-11、10-10、10-9、10-8、10-7 mol/L)ALD处理48 h,或用10-9 mol/L ALD处理不同时间(24 h、48 h、72 h)对HK2细胞CTGF表达的影响,应用RT-PCR法检测CTGFmRNA水平的表达.结果HK2细胞培养48 h后,空白对照组即有CTGF的基础表达,ALD不同浓度组CTGF mRNA的表达均有增加,与空白对照组相比较差异有统计学意义(P＜0.05或P＜0.01);以10-9 mol/L浓度的ALD作用于HK2细胞,随着作用时间的延长,CTGF mRNA的表达量明显增加,与对照组相比较差异有统计学意义(P＜0.05或P＜0.01).结论ALD能够促进HK2细胞CTGF mRNA水平的表达,并具有剂量依赖性和时间依赖性,ALD可能通过促进CTGF的分泌,发挥其促进肾间质纤维化的作用.