TLR4和NF-κB在大鼠肾缺血再灌注损伤中的表达及意义

目的：观察大鼠肾缺血再灌注损伤后Toll样受体4（TLR4）和核因子-κB（NF-kB）的表达变化及其关系.方法：将40只SD大鼠随机等分为假手术组（S组）和肾缺血再灌注损伤组（I/R组）,建立肾缺血再灌注模型.在肾缺血再灌注后24 h切取肾脏.采用免疫组织化学法观察TLR4蛋自及NF-κB蛋白在肾脏组织中的表达变化及其相关性 采用半定肇逆转录-聚合酶链反应（RT-PCR）检测两组肾组织中TLR4、NF-κB的mRNA水平表达变化.结果：TLR4蛋白在S组大鼠肾小管细胞中有少量表达,在I/R组24 h后肾组织中表达明显增加,与S组比较,差异有统计学意义（P〈0.05）.NF-κB P65蛋白在S组大鼠肾小管细胞胞浆和少量胞核中有表达.在I/R组24 h后肾小管细胞胞浆和胞核均可见NF-κB P65明显表达.差异有统计学意义（P〈0.01）.TLR4和NF-κB在肾缺血再灌注损伤组织中的表达具有明显的相关性.I/R组中的TLR4 mRNA、NF-κB mRNA表达均较S组上调,差异有统计学意义（P〈0.05）.结论：大鼠肾缺血冉灌注损伤后,肾组织TLR4和NF-κB P65的表达明显上调,且有明显的相关性.TLR4有可能通过激活NF-κB继而引起下游的炎症因子募集,介导肾缺血再灌注损伤.     

核因子κB在肝缺血再灌注损伤中的表达及意义

我们研究核因子κB(nuclearfactorκB ,NF κB)与肝缺血再灌注损伤的关系 ,报告如下。材料与方法1.动物模型制备 :SD大鼠 (购自武汉大学医学院实验动物中心 ) 4 8只 ,体重 2 2 5～ 2 75 g/只 ,雌雄不拘 ,随机分为2组 :对照组 (2 4只 )和肝缺血再灌注损伤模型组 (2 4只 )。各组内又根据再灌注后不同时间 (1、3、6、2 4h)分为 4小组 ,每小组 6只。肝缺血再灌注损伤模型 :SD大鼠禁食 12h后 ,麻醉后上腹正中切口进腹 ,分离切断肝脏周围韧带 ,解剖肝门 ,显露肝左、中叶血管胆管蒂并用小号无损伤动脉夹将其阻断 ,肝左、中叶转为苍白证实肝血流…     

缺氧诱导因子1α及氧自由基在肾缺血再灌注损伤中的表达

氧自由基参与肾缺血引起的急性肾衰的病理过程。研究发现，缺氧诱导因子1α（HIF-1α）可以增强缺血心肌对缺血缺氧的耐受性。HIF-1α是否参与肾脏缺血再灌注损伤过程及其与氧自由基联系尚未见相关报道。我们通过建立大鼠肾缺血再灌注模型，为临床防治肾缺血再灌注损伤提供依据。     

缺血预处理在移植肾缺血-再灌注损伤中的应用进展

肾移植相对于其他器官移植术后疗效更为显著,但肾缺血-再灌注损伤（IRI）等术后并发症严重影响受者生存率和生存质量,如何减轻移植肾IRI成为了肾移植领域当前的研究重点。目前认为缺血预处理使移植肾适应缺血是预防IRI发展的有效方法之一,但具体机制尚未完全清楚。本文就缺血预处理在IRI中的应用,缺血预处理对移植肾IRI的调控机制,包括细胞水平的调控和细胞内信号通路的调控,以及缺血预处理的临床应用价值和前景进行综述,以期为改善移植肾IRI,提升肾移植受者和移植肾存活率,改善受者生存质量提供参考。     

急性肢体缺血术后再灌注损伤的防治

急性肢体缺血(acute limb ischemia,ALI)是血管外科常见急症,如救治不及时,可致肢体发生不可逆坏死而截肢,甚至危及病人生命.缺血再灌注损伤(ischemia-reperfusioninjury,IRI)是ALI再通术后常见现象,是该病术后并发症发生率和病死率高的主要原因,因此减轻IRI一直是ALI术后处理的重点.     

川芎嗪在大鼠肾脏缺血再灌注损伤中的作用

目的：探讨川芎嗪对大鼠肾脏缺血再灌注损伤的保护作用。方法：观察川芎嗪注射液对大鼠肾脏缺血再灌注损伤后血浆超氧化物歧化酶（SOD）、脂质过氧化物丙二醇（MDA）、内皮素－1（ET－1）的作用及肾小管损伤形态学的影响。结果：川芎嗪治疗组大鼠血浆SOD水平较缺血再灌注组显著升高（P＜0.05),MDA和ET－1水平显著性下降（P＜0.05),肾小管损伤的病理分级显著减轻（P＜0.05)。结论：川芎嗪对肾脏缺血再灌注性损伤具有保护作用。     

香叶木素对小鼠肾缺血再灌注损伤的保护作用

目的 分析香叶木素对小鼠肾缺血再灌注(I/R)损伤是否具有保护作用及其潜在机制。方法 将45只BALB/c雄性小鼠分为假手术组、模型组和香叶木素处理组,通过检测血生化指标和肾组织HE染色损伤评分评价香叶木素对小鼠肾功能的保护作用,通过免疫组化和蛋白免疫印迹检测NF-κB家族关键分子p-P65表达水平变化的情况,通过实时荧光定量聚合酶链式反应检测NF-κB信号通路下游炎症因子的变化情况。结果 与假手术组比较,模型组小鼠血肌酐和尿素氮水平显著升高,小鼠肾组织损伤显著,p-P65以及下游炎症因子表达显著增加。通过香叶木素处理后,可改善肾功能血生化指标,减轻小鼠肾组织损伤,并发现香叶木素可以下调肾I/R损伤中p-P65和相关炎症因子的表达。结论 香叶木素可能通过抑制NF-κB信号通路的激活,从而在小鼠肾I/R损伤中起到保护作用。     

内皮素在急性肾缺血再灌注损伤中作用的实验研究

内皮素在急性肾缺血再灌注损伤中作用的实验研究涂响安余明年谢佛龙赵志毅冯家骅何炳辉彭轼平用放射免疫方法（ＲＩＡ）检测大鼠左肾动脉夹闭６０分钟致急性肾缺血再灌注损伤（ＡＲＲＩ）模型其血浆和肾组织内皮素（ＥＴ）水平变化，以探讨ＥＴ在急性肾缺血再灌注损伤中的...     

NF—κB与脑缺氧、缺血再灌注损伤

核因子NF－κB是一组存在于细胞和病毒中调控许多基因表达的重要转录调节因子之一。当中枢神经系统发生某些病理改变时，NF－κB表现特异的活化。本文综述脑缺氧、缺血再灌注损伤后NF－κB的活化及其可能机制，为进一步研究提供理论依据。     

Toll样受体在肾缺血再灌注损伤中的研究进展题录

肾缺血再灌注损伤(RIRI)造成的急性肾损伤(AKI)在临床上的病死率很高。目前认为Toll样受体(TLR)与其介导的信号通路在其发病机制中占据着重要地位。相关实验表明, 通过抑制TLR及其相关信号通路, 可以抑制炎症反应的发生发展, 减少炎症因子的产生, 从而减轻肾损伤。本文对Toll样受体与其信号通路在RIRI中的作用机制进行综述, 以期为临床治疗RIRI提供新的思路。     

Expression of stromal cell-derived factor 1 in the acute ischemia-reperfusion injury and its relationship with macrophages

Objective To observe the expression of stromal cell-derived factor 1 (SDF-1) in the kidney after ischemic reperfusion injury (IRI), and explore its relationship with macrophage during the IRI kidney. Methods A total of 28 healthy C57BL/6 male mice were used to establish renal IRI model by clamping both pedicles for 35 min followed by reperfusion. Kidney tissue samples were collected at indicated time points. Renal histological changes were estimated. The expression of SDF-1 was determined by immunohistochemistry, ELISA and real-time PCR. After the liposomal clodronate was injected intraperitoneally, the location of CD68 was observed by immunofluorescence. Renal histology and protein expression of SDF-1 were also detected. Results Compared with sham-operated group, classical tubular damage was found in IRI group, accompanied by a large number of inflammatory cells. The expression of total renal SDF-1 peaked on day 1 and decreased to control levels in the following days. SDF-1 in healthy kidney was localized at cortex, but spread to the corticomedullary area of the kidney during IRI. Compared with IRI groups, elimination of macrophage by injection of liposomal clodronate alleviated renal IRI and down-regulated the expressions of CD68 while up-regulating SDF-1. Conclusions SDF-1 expression is up-regulated in IRI kidney and is associated with macrophage. SDF-1 may play a role in the early phase of acute kidney injury and it may be a new marker in diagnosis of AKI.     

热休克蛋白27与缺血-再灌注损伤关系的研究进展

热休克蛋白27是细胞在应激状态下产生的一种小分子蛋白,具有高度保守性,在抗缺血-再损伤中发挥重要作用.随着器官移植技术的不断提高,心脏、肝脏、肾脏等器官移植越来越多,器官的缺血-再灌注损伤成为移植物存活与否的一个不可忽视的环节,研究内源性的器官保护机制,显得尤为重要.本文就热休克蛋白27与缺血-再灌注损伤的关系作一综述.     

雷帕霉素对肾脏缺血再灌注损伤的影响

移植肾在经历缺血再灌注损伤后可以产生不同程度的损伤,而雷帕霉素作为临床上常用的免疫抑制剂,又可对再灌注的移植肾产生多方面的影响.本文就雷帕霉素对肾脏缺血再灌注损伤的影响作一综述.     

小鼠急性缺血-再灌注肾损伤模型的建立及体会

目的观察应用小鼠制备急性缺血一再灌注性肾损伤模型的效果。方法应用微型动脉夹夹闭小鼠双侧肾动脉制备急性缺血一再灌注肾损伤模型,其中两组分别于术后24h和48h后处死观察肾功能及肾脏病理变化．另一组观察其病情及存活情况14天。结果各次造模成功率均达85％以上;术后24h及48h实验组血清肌酐（Scr）和血尿素氮（BUN）水平明显升高,与对照组比较差异有统计学意义（P〈0．01）：实验组肾脏外观出现典型“大白肾”表现,镜下出现典型急性肾小管坏死表现,并有较多炎症细胞浸润,肾小管组织学评分与对照组比较差异有统计学意义（P均〈0．01）;实验组在观察期间逐渐出现典型急肾衰竭表现,至14天末,死亡率达91．7％,而对照组全部正常存活。结论应用微型动脉夹夹闭小鼠双侧肾动脉可制备稳定急性缺血一再灌注肾损伤模型,而且成功率较高。     

Prevention and management of acute kidney injury in the perioperative patient

Acute kidney injury does not just describe a set of abnormal blood results, it is a medical emergency and biomarker of acute illness. It is a common and costly surgical complication, increasing perioperative morbidity and mortality. Key to managing these patients successfully and reducing the massive burden AKI and its sequelae has on the NHS is rapid identification and intervention, not just of patients who have developed AKI but also those at risk. Once identified, robust strategies must be in place to ensure appropriate treatment, identification and treatment of underlying causes, escalation of care and specialist referral if indicated, take place promptly.     

Prevention and management of acute kidney injury in the perioperative patient

Acute kidney injury (AKI) does not just describe a set of abnormal blood results, it is also a biomarker of acute illness. It is a common and costly surgical complication, increasing perioperative morbidity and mortality. Key to managing these patients successfully and reducing the massive burden AKI and its sequelae is rapid identification and intervention, not just of patients who have developed AKI but also those at risk. Once identified, robust strategies must be in place to ensure appropriate treatment, identification and treatment of underlying causes, escalation of care and specialist referral if indicated, take place promptly.     

Role of OMA1 in lipopolysaccharide-induced acute kidney injury

Objective To investigate the role of OMA1 in acute kidney injury (AKI) induced by lipopolysaccharide (LPS). Methods OMA1 wild-type and knocked out mice (8 week old) were injected with 10 mg/kg body weight of LPS. The model was confirmed by testing mouse serum creatinine and blood urea nitrogen. The apoptosis in mouse kidney cortex was examined by TUNEL staining and cleaved caspase 3. In vitro, in humam kidney proximal tubular cells (HK2) were knocked down OMA1 by transfecting OMA1 shRNA, with the scramble shRNA being used as negative control of transfection. HK2 cells were cultured with 5 μg/ml of LPS for 24 hours to induce apoptosis. DAPI staining of cells and caspase-3 activity were applied to test apoptosis. The images of mitochondria in cells were obtained by transfection of mito-green plasmid and OMA1 shRNA. Western blotting was used to exam the OMA1 and Cytochrome C expressions. Results Compared with OMA1 KO mice, LPS induced more severe AKI of WT mice with higher Scr [(97.2±26.5) μmol/L vs (53.0±17.7) μmol/L, P＜0.05] and BUN [(43.3±13.7) mmol/L vs (29.7±7.7) mmol/L, P＜0.05]. Moreover, there were more apoptosis cells in kidney cortex in WT mice than in OMA1 KO mice [(75.4± 26.1)/mm2 vs (38.3± 14.4)/mm2, P＜0.05]. About 46% of OMA1 expressions in HK2 cells were inhibited by OMA1 shRNA transfection (P＜0.05). Further, OMA1 shRNA cells with LPS stimulation had decreased mitochondria fragmentation [(29.8±10.9)% vs (43.2±6.8)% , P＜0.05], Cytochrome C release [(37.0±12.3)% vs (76.0±26.2)%, P＜0.05], and cell apoptosis [(13.2±3.9)% vs (25.0±7.1)%, P＜0.05] as compared with control cells. Conclusion Knockdown of OMA1 alleviated septic AKI through inhibition of cell apoptosis, mitochondria fragmentation, and Cytochrome C release.     

骨髓间充质干细胞对大鼠急性缺血再灌注肾损伤的修复作用

目的 观察骨髓间充质干细胞(BMSCs)移植对大鼠缺血再灌注(IR)急性肾损伤(AKI)的修复作用.方法 将50只SD大鼠随机分为3组,干细胞移植组(IR+ BMSCs组)18只,生理盐水注射组(IR组)18只,正常组(Sham组)14只.分离及培养大鼠BMSCs到第3代,建立大鼠缺血再灌注AKI模型,24h后移植干细胞;检测血清及尿肌酐值,并以细胞核增殖抗原(Ki-67)进行肾脏免疫组织化学观察.结果 BMSCs培养后,得到纯度高、生长状态良好的第3代(P3) BMSCs.造模后第4天,细胞治疗组血清肌酐浓度为(37.34±4.22) μmol/L,尿肌酐水平为(22.75±6.23) μmol,而生理盐水注射组血清肌酐浓度为(238.34±32.42) μmol/L,尿肌酐水平为(7.68±1.03) μmol,两组比较差异有统计学意义(P＜0.01);组织切片学观察,与对照组比较,IR+ BMSCs组肾小管结构改善及上皮细胞数量增多、刷状缘部分恢复,Millar法评分IR+ BMSCs组为0.799±0.023,IR组为2.798±0.055,两组比较差异有统计学意义(P＜0.01).以Ki-67进行免疫组织化学IR+ BMSCs组有大量增殖抗原阳性表达.结论 BMSCs对缺血再灌注性AKI有良好的修复作用.     

自噬和ASPPs在老年大鼠急性肾损伤早期的表达

目的观察自噬相关蛋白和p53凋亡刺激蛋白(ASPPs)在肾损伤早期的表达变化,初步探讨自噬相关蛋白和ASPPs是否可能成为老年大鼠AKI早期生物标志物。 方法建立顺铂致AKI青年与老年大鼠模型。雄性SD老年大鼠随机分为假手术组(Sham),顺铂模型组,同时设数量匹配的雄性SD青年大鼠为对照;模型组大鼠一次性腹腔注射顺铂4 mg/kg,Sham组相同途径注射生理盐水4 ml/kg;在给药12 h、1 d、3 d、5 d、7 d时检测大鼠Scr、BUN;光镜观察大鼠肾脏病理变化;透射电镜观察大鼠肾小管上皮细胞超微结构变化及自噬体的情况;免疫印迹法检测肾脏组织Beclin 1、溶酶体相关膜蛋白2(LAMP2)、p62、p53及ASPP抑制物(iASPP)和ASPP1表达情况。 结果顺铂诱导12 h后,与Sham组比较,青年与老年大鼠Scr无明显变化(P>0.05);电镜观察到大鼠肾小管上皮细胞自噬体出现而且数量显著增多;老年大鼠肾组织Beclin 1、p62、LAMP-2和p53表达水平明显升高(P<0.05),iASPP表达水平明显降低(P<0.05),并且老年大鼠肾组织Beclin 1、LAMP-2和p53变化时间早于青年大鼠(P<0.05)。 结论自噬和ASPPs在老年大鼠AKI发生早期即可出现,在Scr开始升高前,反应性自噬已经启动。自噬相关蛋白和ASPPs有望成为AKI早期的损伤标志物,可能是AKI早期干预的新靶点,但仍需更深入的研究。