BALB/c小鼠肾缺血再灌注损伤模型的建立与评价

背景：对于一些药物研究,小鼠是理想的造模工具,但由于小鼠耐受性相对较差,肾脏及肾蒂小且难于寻找,容易增加实验误差,导致造模失败。 目的：探讨BALB/c小鼠肾缺血再灌注损伤模型的建立方法,评价肾脏缺血时间对肾缺血再灌注损伤的影响。 方法：采用微型动脉夹夹闭小鼠双侧肾蒂的方法建立雄性BALB/c小鼠肾缺血再灌注损伤模型,根据肾缺血时间不同分为    0 min组(对照组)、30 min组、35 min组、45 min组,肾再灌注后24 h观察肾功能和肾脏病理组织学的变化,比较不同的肾脏缺血时间对上述指标的影响;观察45 min组小鼠肾缺血再灌注损伤后的生存率。 结果与结论：模型成功率95.9%,与对照组相比,肾缺血30 min组、35 min组和45 min组再灌注后24 h血清肌酐、尿素氮和肾脏病理组织学评分均升高,肾缺血45 min组生存率明显下降,差异均有显著性意义(P < 0.05)。结果提示,应用微型动脉夹夹闭小鼠双侧肾蒂的方法可制备稳定肾缺血再灌注损伤模型,雄性小鼠肾缺血35~45 min是造模较为理想的肾缺血时间,所得模型效果满意。     

大鼠急性肾损伤模型的建立及意义

目的探讨建立大鼠缺血再灌注急性肾损伤模型。方法采用切除大鼠右肾,左肾蒂夹闭45分钟,再灌注2、6、12、24 h测定血尿素氮(BUN)、肌酐(Scr)的水平,观察肾组织病理形态学改变,TUNEL法检测肾小管凋亡细胞,确定大鼠缺血再灌注急性肾损伤模型。结果 IRI组BUN、Scr在再灌注后2h开始上升,在24 h达到高峰;肾组织24 h后肾小管上皮细胞凋亡、坏死脱落,小管可见细胞碎片及管型,小管排列稀疏、紊乱,管腔明显扩张,肾小球无明显改变;与Sham组相比,IRI各亚组肾小管凋亡细胞指数与损伤评分,差异有统计学意义(P0.05),且随缺血再灌注时间延长及肾功能损伤的加重,肾小管凋亡细胞指数与损伤评分增加。结论切除大鼠右肾、夹闭左肾动脉的方法可成功建立缺血再灌注急性肾损伤动物模型。     

丝裂素活化蛋白激酶亚类在急性缺血/再灌注肾损伤修复过程中的变化

目的:研究急性缺血/再灌注肾损伤修复时丝裂素活化蛋白激酶(MAPKs)亚类细胞外信号调节激酶(ERK)、c-Jun氨基末端激酶(JNK)的变化,探讨其在肾损伤修复中的意义。方法:夹闭大鼠双侧肾蒂造成急性缺血/再灌注肾损伤模型,应用电镜观察肾组织形态学改变,免疫组化法检测肾组织增殖细胞核抗原(PCNA)表达,采用特异性底物磷酸化结合免疫沉淀法测定肾组织的ERK、JNK活性。结果:急性缺血后再灌注2h肾小管上皮细胞腔面微绒毛重新出现,肾小管PCNA阳性细胞开始增加;缺血45min使ERK活性降低,再灌注5minERK活性完全恢复;缺血45min对JNK活性无影响,再灌注5minJNK活性增加并持续到再灌注2h。结论:MAPKs活性变化,特别是JNK活性增加参与了急性缺血/再灌注肾损伤早期细胞修复过程。     

缺血再灌注损伤对小鼠肾脏血管内皮生长因子受体3表达的影响

研究肾脏缺血再灌注( ischemia-reperfusion, IR)对小鼠肾脏血管内皮生长因子受体3( vascular endothelial growth factor receptor 3, VEGFR3)表达的影响。方法30只雄性C57BL/6小鼠随即分为5组：假手术组( Sham组)﹑缺血再灌注0﹑6、12﹑24 h组( IR 0 h组﹑IR 6 h组﹑IR 12 h组﹑IR 24 h组),每组6只。缺血再灌注组用无创性动脉夹夹闭左侧肾带,置于32℃温箱后1 h松开血管夹,随后切除右肾。 Sham组操作同上,但不夹闭左侧肾蒂。再灌注0﹑6﹑12﹑24 h后处死小鼠,收集肾脏及小鼠外周血标本。测定血肌酐(Cr)和尿素氮(BUN)水平。 PAS染色后显微镜下观察肾脏病理学变化, Western印迹和免疫组织化学法检测肾脏组织血管内皮生长因子受体3的表达。结果与Sham组相比较,再灌注0 h时小鼠血肌酐和尿素氮无明显升高,但缺血再灌注6﹑12﹑24 h后血肌酐和尿素氮呈显著性上升。 IR各组肾组织病理损伤也随着再灌注时间的延长而逐渐加重,可见肾小管上皮细胞明显肿胀坏死,蛋白管型形成,刷状缘脱落。随着缺血再灌注时间的进展, VEGFR-3蛋白表达量增加,免疫组织化学染色结果显示VEGFR3主要分布在肾脏皮质髓质交界处的肾小管。结论 VEGFR3在肾脏缺血再灌注损伤后表达量上升,且表达部位主要分布于肾脏皮髓质交界处的肾小管,因此VEGFR3可能参与调控肾脏缺血再灌注损伤。     

脑缺血再灌注后小鼠海马神经前体细胞的增殖

目的:研究小鼠脑缺血再灌注后海马神经前体细胞的增殖。方法:采用无创微动脉夹夹闭小鼠双侧颈总动脉0·5h的方法制作脑缺血再灌注动物模型。分正常对照组、假手术组、实验组小鼠脑缺血再灌注后1、3、5、7、14、21、28d组,共7个时相点。各组小鼠于各时相点处死前注射5-溴脱氧尿苷嘧啶(BrdU)。处死后取出脑组织制备石蜡切片进行Nissl染色,BrdU免疫组织化学显色,观察脑组织缺血后的病理变化并比较缺血后不同时相点神经前体细胞增殖的差异。同时从小鼠心采血进行血气分析。结果:夹闭小鼠双侧颈总动脉的脑缺血再灌注模型可致PO2明显降低,PCO2明显升高,大脑皮质出现了缺血的形态学变化。小鼠海马BrdU 细胞在脑缺血再灌注后1d开始增加,7d达到高峰,28d恢复正常水平。结论:夹闭双侧颈总动脉是致小鼠脑缺血再灌注损伤的理想模型。小鼠脑缺血再灌注损伤能激活海马神经前体细胞的增殖,并在术后7d达峰值。     

肾缺血再灌注对大鼠室旁核电活动的影响

为了观察肾缺血-再灌注对室旁核电活动的影响,探讨中枢神经活动在急性缺血再灌注肾损伤发生、发展中的作用和机制。本实验通过夹闭大鼠一侧肾动脉30min、再灌注30min的肾缺血再灌注模型,同步动态记录肾动脉夹闭致缺血-再灌注性肾损伤大鼠室旁核的放电频率、放电周期、放电积分等参数的变化。结果显示:(1)与对照组相比,肾缺血瞬间室旁核电活动受到抑制,缺血5min后电活动逐渐增强并维持在较高水平;(2)去夹闭再灌流瞬间放电立即减少,再灌注5min后放电又逐渐增强,并于再灌注20min～25min时室旁核放电达最强,与缺血前及缺血中各个观测点相比,均有显著性差异(P<0.05～0.01)。上述结果提示肾脏在缺血和缺血-再灌注过程中可引起室旁核电活动发生周期性变化,其中再灌注后对室旁核电活动的影响较缺血的影响更为明显。     

促红细胞生成素对急性肾损伤中干细胞因子及其受体表达的影响

背景：细胞因子在减轻肾脏缺血再灌注损伤中的作用日益受到重视,干细胞因子具有造血系统以外的器官保护作用。目的：探讨大鼠急性肾损伤模型中干细胞因子及其受体c-Kit表达的变化和促红细胞生成素预处理对其表达的影响。方法：成年雄性Wistar大鼠34只,采用夹闭双侧肾蒂建立缺血再灌注模型,缺血45 min后再灌注24 h,随机分为假手术组(n=10)、缺血再灌注组(n=12)和促红细胞生成素组(n=12),促红细胞生成素组于造模前2 h一次性尾静脉注射重组人促红细胞生成素5 000 U/kg。免疫组化及图像分析技术检测各组肾组织中干细胞因子及c-Kit的表达变化,测定血清肌酐和尿素氮水平,苏木精-伊红染色观察肾组织病理学改变并计算肾小管损伤积分。结果与结论：干细胞因子及c-Kit在肾组织中的表达仅限于肾小管区域。与假手术组比较,干细胞因子和c-Kit在缺血再灌注组和促红细胞生成素组的表达均明显增高(P < 0.05),促红细胞生成素组干细胞因子表达高于缺血再灌注组(P < 0.01),但两组c-Kit表达差异无显著性意义(P > 0.05);促红细胞生成素组血清肌酐与尿素氮水平明显低于缺血再灌注组(P < 0.05),但高于假手术组(P < 0.05)。与缺血再灌注组比较,促红细胞生成素组肾组织病变减轻。说明缺血再灌注导致急性肾损伤发生时干细胞因子及c-Kit表达升高,而促红细胞生成素对急性肾损伤的保护作用可能与上调干细胞因子及c-Kit表达有关。中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

P53在急性肾损伤小鼠肾脏的表达及其与细胞凋亡的关系

 目的：探讨缺血再灌注致急性肾损伤（AKI）小鼠肾脏不同部位P53的表达及其与细胞凋亡的关系。方法：采用随机对照动物实验方法,将18只小鼠随机分为假手术组、AKI组及P53抑制剂pifithrin-alpha（PFT-α）组,每组6只。采用双侧肾蒂夹闭45 min后松开动脉夹的方法建立小鼠AKI模型,PFT-α组于建模前5 min腹腔注射PFT-α  2.2 mg/kg,并于建模后48 h采血检测尿素氮和肌酐,取肾脏组织行HE染色观察肾组织病理学变化,采用Western blotting法测定P53蛋白含量,免疫荧光法确定肾脏不同部位P53的表达,TUNEL法检测肾脏细胞凋亡,免疫组化方法检测肿瘤坏死因子受体（TNFR）、caspase-3及Bcl-2蛋白水平 。结果：（1）PFT-α组和AKI组小鼠血尿素氮和肌酐水平均明显高于假手术组,而PFT-α组与AKI组比较血尿素氮和肌酐水平明显降低（P<0.05）;肾组织HE染色显示假手术组肾组织细胞形态完整,排列整齐,无明显病理改变,AKI组肾小管上皮细胞刷状缘脱落、空泡及滴状变性,皮髓质间有明显淤血带;PFT-α组肾小管上皮部分刷状缘脱落消失,空泡及滴状变性减轻,皮髓质间无明显淤血带;（2）假手术组小鼠肾脏有少量P53表达且未检测到凋亡细胞,而AKI组小鼠缺血再灌注48 h后P53蛋白水平及凋亡细胞明显增加（P<0.05）,并且均主要定位在肾皮质,PFT-α组较AKI组小鼠肾脏P53蛋白含量及凋亡细胞指数均减少（P<0.05）;（3）与假手术组相比,AKI组小鼠肾脏TNFR蛋白及caspase-3蛋白水平升高（P<0.05）,Bcl-2蛋白水平下降（P<0.05）,PFT-α组较AKI组小鼠肾脏TNFR蛋白及caspase-3蛋白水平降低（P<0.05）,Bcl-2蛋白水平升高（P<0.05）。结论：急性肾损伤时,肾组织P53表达增加,主要定位于皮质,通过调控凋亡蛋白Bcl-2和TNFR的水平,促进caspase-3的释放,从而介导细胞凋亡。     

大鼠缺血性肾损伤的细胞聚合糖性死亡与外源性凋亡

目的探讨大鼠肾缺血再灌注所致急性肾损伤时细胞聚合糖性死亡与外源性凋亡。方法应用免疫印迹技术、免疫组织化学染色技术以及光学和电子显微镜技术对缺血60min再灌注24h的大鼠肾组织进行观察和分析。结果免疫印迹分析结果表明,与sham组比较肾缺血再灌注后(AKI组)肾组织PARP-1、caspase-3和TNFRα表达增强。PARP-1、caspase-3免疫组化染色阳性细胞出现在缺血再灌注损伤肾组织,主要分布于肾小管,皮质和髓质外带的肾小管出现了大面积细胞坏死,表现为细胞肿胀,空泡形成,崩解脱落。在髓质外带肾小管坏死细胞之间存在着较多的凋亡细胞,细胞皱缩,核固缩。电镜下坏死细胞肿胀,细胞器也肿胀,崩解消失。凋亡细胞皱缩,界限清楚,核染色质固缩边聚。结论大鼠肾缺血60 min再灌注24 h部分肾小管上皮细胞发生聚合糖性死亡和外源性凋亡。     

丝裂素活化蛋白激酶亚类在急性缺血／再灌注肾损伤修复过？…

目的 研究急性缺血／再灌注肾损伤修复时丝裂素活化蛋白激酶（ＭＡＰＫｓ）亚类细胞外信号调节激酶（ＥＲＫ）、ｃ－Ｊｕｎ氨基末端激酶（ＪＮＫ）的变化,探讨其在肾损伤修复中的意义。方法 夹闭大鼠双侧肾蒂造成急性缺血／再灌注肾损伤模型,应用电镜观察肾组织形态学改变,免疫组化法检测肾组织增殖细胞核抗原（ＰＣＮＡ）表达,采用特异性底物磷酸化结合免疫沉淀法测定肾组织的ＥＲＫ、ＪＮＫ活性。结果 急性缺血后再灌注２ｈ     

甘油致BALB/C小鼠急性肾损伤后肾脏观察

目的 观察甘油致小鼠急性肾脏损伤后肾脏修复的情况.方法 肌肉注射甘油致小鼠发生急性肾脏损伤后出取肾脏组织,于损伤后的不同时间点进行肾脏组织形态学检测;同时,应用免疫组织化学技术检测核增殖抗原在肾脏组织的表达情况,分析肾脏损伤后的修复情况.结果 肌肉注射甘油3 d后小鼠开始出现急性肾脏损害的表现.主要表现为肾脏肿胀,肾脏...     

Detection and evaluation of renal biomarkers in a swine model of acute myocardial infarction and reperfusion

The prevalence of type 1 cardiorenal syndrome (CRS) is increasing and strongly associated with long-term mortality. However, lack of reliable animal models and well-defined measures of renoprotection, made early diagnosis and therapy difficult. We previously successfully established the swine acute myocardial infarction (AMI) model of ischemia-reperfusion by blocking left anterior descending branch (LAD). Reperfusion was performed after 90-minute occlusion of the LAD. AMI was confirmed by ECG and left ventricular angiography (LVG). Then those 52 survived AMI reperfusion swine, including ventricular fibrillation-cardiac arrest after restoration of blood flow, were randomly divided into four groups (four/group) according to different interventions: resuscitation in room temperature, resuscitation with 500 ml saline in room temperature, resuscitation with 4°C 500 ml saline and normal control (with no intervention of resuscitation). Each group was further observed in four groups according to different time of resuscitation after ventricular arrhythmias: 1, 3, 5, 10-minute reperfusion after ventricular arrhythmias. Plasma and random urine were collected to evaluate renal function and test renal biomarkers of acute kidney injury (AKI). Our swine AMI model of ischemia-reperfusion provoked subclinical AKI with the elevation of the tubular damage biomarker, NGAL, IL-18 and L-FABP. Renal damage rapidly observed after hemodynamic instability, rather than observation after several hours as previously reported. The increasing rate of biological markers declined after interventions, however, its impact on the long-term prognosis remains to be further studied. These data show that elevation of tubular damage biomarkers without glomerular function loss may indicate appropriate timing for effective renoprotections like hypothermia resuscitation in type 1 CRS.     

肾缺血-再灌注损伤引起的脑生化改变

目的:观察血清及脑脊液生化成分变化，初步探讨肾缺血-再灌注损伤对脑的影响。 方法: 20只健康新西兰兔随机分为对照组和肾缺血再灌注组（IR组），检测和比较血清和脑脊液中肌酐（Cr）、尿素氮(UN)、Na+ 、Ca2+、一氧化氮（NO）代谢产物含量及脑组织总一氧化氮合酶（NOS）活力。 结果: 在肾缺血再灌注后24 h，IR组血清和脑脊液中的UN、Cr和NO均显著高于对照组（P<0.05），Na+明显低于对照组；IR组血清Ca2+显著低于（P<0.05）、而脑脊液中Ca2+显著高于对照组（P<0.05）。IR组脑组织中NO2-/NO3-含量和NOS活力均明显高于对照组（P<0.05）。 结论: 肾缺血再灌注损伤不仅影响血清成分，而且还可引起脑生化的改变；NO可能参与了肾缺血-再灌注损伤对脑功能的影响。     

高压氧对短暂前脑缺血小鼠海马脑红蛋白表达的影响

目的 观察小鼠脑缺血再灌注以及高压氧治疗后海马脑红蛋白表达的变化,探讨高压氧对脑红蛋白表达的影响及其意义.方法 雌性C57B/6N小鼠随机分为假手术对照组、缺血再灌注48h组(48h I/R组)及缺血再灌注48h加高压氧治疗组(48h HBO组).对各组小鼠脑组织神经元进行HE及免疫组织化学染色、图像学分析及统计学处理.结果 48h I/R组及48h HBO组的脑红蛋白表达均高于假手术组(P＜0.01和P＜0.001),48hHBO组吸光度高于48h I/R组(P＜0.01).结论 高压氧对缺血性脑损伤有显著治疗作用,提高脑红蛋白的表达水平可能是其治疗的机制之一.     

Effect of combination of vitamin E and umbilical cord-derived mesenchymal stem cells on inflammation in mice with acute kidney injury

Background: The objective of this study is to investigate the effect of combination of umbilical cord-derived mesenchymal stem cell (UC-MSC) and vitamin E (VitE) on inflammation in mice with acute kidney injury (AKI).

Methods: UC-MSCs were isolated from pregnant wistar mice and cultured. A total of 90 female wistar mice were randomly divided into control group, AKI group, AKI?+?VitE group, AKI?+?UC-MSC group, and AKI?+?VitE?+?UC-MSC group (18 mice in each group) which were given no treatment, normal saline, VitE, UC-MSC, and VitE?+?UC-MSC, respectively. The renal pedicles on both sides were clipped for 50?min with micro-artery clips to induce AKI. Six mice were sacrificed at days 1, 3, and 7, while blood and kidney tissues were collected to detect levels of blood urea nitrogen (BUN) and creatinine (Scr). Kidney tissues were stained by HE staining to observe pathological changes; levels of interleukin-lβ, TNF-α, interleukin-10, and β-FGF were measured by ELISA.

Results: Compared with the control group, AKI mice showed higher levels of serum BUN and Scr, tubular swelling and necrosis suggesting that AKI model was successfully established. Mice in AKI?+?VitE group, AKI?+?UC-MSC group, and AKI?+?VitE?+?UC-MSC presented better renal function than mice of AKI group. Mice from AKI?+?VitE?+?UC-MSC group showed the best renal function with the least renal tubular injury (p?p?p?Conclusions: Combination of UC-MSC and VitE significantly inhibited inflammatory reaction in kidney through the regulation of inflammatory cytokines in the microenvironment of kidney with AKI. Combination of UC-MSC and VitE presented therapeutic effect on AKI than the single use of UC-MSC or VitE.     

Managing stress and anxiety through qigong exercise in healthy adults: a systematic review and meta-analysis of randomized controlled trials

Background

Renal ischemia-reperfusion injury (IRI) increases the rates of acute kidney failure, delayed graft function, and early mortality after kidney transplantation. The pathophysiology involved includes oxidative stress, mitochondrial dysfunction, and immune-mediated injury. The anti-oxidation, anti-apoptosis, and anti-inflammation properties of baicalin, a flavonoid glycoside isolated from Scutellaria baicalensis, have been verified. This study therefore assessed the effects of baicalin against renal IRI in rats.

Methods

Baicalin was intraperitoneally injected 30 min before renal ischemia. Serum and kidneys were harvested 24 h after reperfusion. Renal function and histological changes were assessed. Markers of oxidative stress, the Toll-like receptor (TLR)2 and TLR4 signaling pathway, mitochondrial stress, and cell apoptosis were also evaluated.

Results

Baicalin treatment decreased oxidative stress and histological injury, and improved kidney function, as well as inhibiting proinflammatory responses and tubular apoptosis. Baicalin pretreatment also reduced the expression of TLR2, TLR4, MyD88, p-NF-κB, and p-IκB proteins, as well as decreasing caspase-3 activity and increasing the Bcl-2/Bax ratio.

Conclusions

Baicalin may attenuate renal ischemia-reperfusion injury by inhibiting proinflammatory responses and mitochondria-mediated apoptosis. These effects are associated with the TLR2/4 signaling pathway and mitochondrial stress.     

促红细胞生成素对急性肾损伤大鼠肾小管细胞凋亡和p-MAPKAPK-2表达的影响

目的:探讨促红细胞生成素(EPO)对急性肾损伤大鼠肾小管细胞凋亡和MAPKAPK-2水平的影响.方法:30只SD雄性大鼠分为假手术组、模型组、EPO治疗组,假手术组仅分离肾被膜后逐层关闭腹腔;模型组采用缺血再灌注法建立急性肾损伤模型;EPO治疗组为急性肾损伤大鼠腹腔注射EPO 50 U/kg,每周3次,共6周,其余大鼠...     

Renal cholesterol accumulation: a durable response after acute and subacute renal insults

Proximal tubular cholesterol levels rise within 18 hours of diverse forms of acute renal tubular injury (eg, myoglobinuria, ischemia/reperfusion, urinary tract obstruction). These increments serve to protect against further bouts of tubular attack (so-called "acquired cytoresistance"). Whether these cholesterol increments are merely transitory, or persist into the maintenance phase of acute renal failure (ARF), has not been previously defined. Furthermore, whether subacute/insidious tubular injury [eg, cyclosporine A (CSA), tacrolimus toxicity], nontubular injury (eg, acute glomerulonephritis), or physiological stress (eg, mild dehydration) impact renal cholesterol homeostasis have not been addressed. This study sought to resolve these issues. Male CD-1 mice were subjected to glycerol-induced ARF. Renal cortical-free cholesterol (FC) and cholesterol ester (CE) levels were determined 3, 5, 7, or 14 days later, and the values contrasted to prevailing blood-urea nitrogen concentrations. The impact of 40 minutes of unilateral renal ischemia plus reflow (3 to 6 days) on mouse cortical FC/CE content was also assessed. Additionally, FC/CE levels were measured in rat renal cortex either 10 days after CSA or tacrolimus therapy, or 48 hours after induction of nephrotoxic serum nephritis. Finally, the impact of overnight dehydration on mouse renal cortical/medullary FC/CE profiles was determined. Compared to sham-treated animals, glycerol, CSA, tacrolimus, ischemia-reperfusion, and nephrotoxic serum each induced dramatic CE +/- FC elevations, rising as much as 10x control values. In the glycerol model, striking correlations (r 相似文献   

Beagle犬药物诱导急性肾损伤模型的研究

目的 建立顺铂诱导的Beagle犬急性肾损伤模型,为开展肾毒性生物标志物研究奠定基础.方法 通过对Beagle犬单次静脉注射3、4和5 mg/kg顺铂,探讨采用顺铂建立急性肾小管损伤模型的给药方案.密切观察动物给药后临床症状,测定不同时间点的传统血清及尿液新型肾损伤生物标志物的动态变化,在试验结束时解剖动物并进行肾脏组...