缺血后处理下调脑缺血再灌注大鼠白细胞介素-1β和肿瘤坏死因子-α表达

目的 探讨缺血后处理(ischemic postconditioning,IP)对局灶性脑缺血再灌注大鼠白细胞介素-1β(interleukin- 1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达的影响.方法 110只成年健康雄性Sprague-Dawley大鼠随机分为假手术组(n=10)、缺血再灌注组和IP组,后2组根据再灌注时间再分为6、12、24、48和72 h亚组(每亚组n=10).采用大脑中动脉线栓法建立局灶性脑缺血再灌注模型.IP方法为在大脑中动脉闭塞2h后实施再灌注15 s/缺血15 s,反复3次.对各组大鼠进行神经行为学评分,2,3,5-氯化三苯基四氮唑染色测定脑梗死体积,免疫组化法检测脑组织TNF-α和IL-1β蛋白表达,原位杂交法检测IL-1β和TNF-α mRNA表达.结果 与缺血再灌注组比,IP组神经行为学评分显著降低(P均＜0.05),脑梗死体积显著缩小(P均＜0.05).假手术组额顶叶皮质IL-1β、TNF-α蛋白和mRNA表达微弱;缺血再灌注组IL-1β、TNF-α蛋白和mRNA在额顶叶皮质大量表达,6h开始上调,24 h达高峰(与其他时间点比较,p均＜0.05),之后逐渐下降;IP组具有相同的动态变化趋势,且各时间点表达量均显著低于缺血再灌注组(P均＜0.05).结论 IP可显著下调脑组织IL-1β和TNF-α表达,缩小缺血再灌注大鼠脑梗死体积,提示IP可通过抑制脑组织缺血再灌注后炎症反应发挥神经保护作用.     

落新妇甙对热缺血再灌注损伤肝脏肿瘤坏死因子α与白细胞介素-10表达的影响

目的 观察落新妇甙对热缺血再灌注(I/R)损伤肝脏Th1、Th2型细胞因子表达的影响. 方法 C57BL/6小鼠随机分为4组:假手术组、模型组,落新妇甙小剂量(10 mg/kg)干预组和大剂量(40mg,/kg)干预组.干预组小鼠于缺血前24h和1 h分别给予10mg/kg或40mg/kg的落新妇甙腹腔注射,建立70%部分肝I/R模型.收集肝组织标本,Western blot检测肝组织中肿瘤坏死因子(TNF)α,白细胞介素(IL)-10蛋白含量,RT-PCR检测肝组织TNF α、IL-10 mRNA的水平.多组间比较采用单因素方差分析,两两比较采用SNK法.结果 与I/R模型对照组比较,小、大剂量落新妇甙干预组肝组织中TNF α蛋白表达水平依次降低,与TNF α mRNA的半定量RT-PCR结果 相符(相对表达量分别为1.74±O.12与1.56±0.09、0.82±0.12,P值均<0.05);而IL-10蛋白表达水平依次升高,与IL-10 mRNA的半定量RT-PCR结果 相符(相对表达量分别为0.96±0.08与1.11±0.17、1.47±0.08,P值均<0.05).结论 落新妇甙干预能抑制I/R损伤肝组织中Th1型细胞因子TNFα的高表达,同时促进Th2型细胞因子IL-10的表达.     

落新妇甙对热缺血再灌注损伤肝脏肿瘤坏死因子α与白细胞介素-10表达的影响

Objective To investigate the effects of astilbin on the expressions of TNF α and IL-10 during liver warm ischemia-reperfusion injury. Methods C57BL/ 6 mice were randomly divided into 4 groups (n = 8): sham-operated group (Sham), model control group(I/R), low dosage of astilbin treatment group (10 mg/kg) and high dosage of astilbin (40 mg/kg) treatment group. The treatment group mice were intraperitoneally injected with 10 or 40 mg/kg astilbin 24 hours and one hour before Ischemia, the hepatic ischemia-reperfusion model were thus established. After jn90 of min ischemia and 6 h reperfusion of the partial hepatic lobe, the expressions of TNF α and IL-10 in liver tissues collected from the experimental groups were detected by Western blot and semiquantitative RT-PCR. Results The expression of TNF a protein in liver tissues gradually decreased in treatment groups (low and high dosages of astilbin treatment groups) as compared to the I/R model control group. Similar results were observed in the mRNA expressions of these genes as determined by semiquantitative RT-PCR (P < 0.05 for low dosage group; P < 0.01 for high dosage group). Compared with the I/R model control group, the expression of IL-10 was increased in both treatment groups (low dosage group P < 0.05; large dosage group P < 0.01). Conclusion Treatment with astilbin decreases TNF α expression but induces IL-10 expression in liver during warm ischemia-reperfusion injury.     

落新妇甙对热缺血再灌注损伤肝脏肿瘤坏死因子α与白细胞介素-10表达的影响

Objective To investigate the effects of astilbin on the expressions of TNF α and IL-10 during liver warm ischemia-reperfusion injury. Methods C57BL/ 6 mice were randomly divided into 4 groups (n = 8): sham-operated group (Sham), model control group(I/R), low dosage of astilbin treatment group (10 mg/kg) and high dosage of astilbin (40 mg/kg) treatment group. The treatment group mice were intraperitoneally injected with 10 or 40 mg/kg astilbin 24 hours and one hour before Ischemia, the hepatic ischemia-reperfusion model were thus established. After jn90 of min ischemia and 6 h reperfusion of the partial hepatic lobe, the expressions of TNF α and IL-10 in liver tissues collected from the experimental groups were detected by Western blot and semiquantitative RT-PCR. Results The expression of TNF a protein in liver tissues gradually decreased in treatment groups (low and high dosages of astilbin treatment groups) as compared to the I/R model control group. Similar results were observed in the mRNA expressions of these genes as determined by semiquantitative RT-PCR (P < 0.05 for low dosage group; P < 0.01 for high dosage group). Compared with the I/R model control group, the expression of IL-10 was increased in both treatment groups (low dosage group P < 0.05; large dosage group P < 0.01). Conclusion Treatment with astilbin decreases TNF α expression but induces IL-10 expression in liver during warm ischemia-reperfusion injury.     

缺氧诱导因子-1α在鼠肺缺血再灌注损伤中的表达与意义

目的:探讨缺氧诱导因子-1α(hypoxia inducible factor-1 alpha,HIF-1α)对鼠肺缺血再灌注损伤是否具有保护作用及可能的机制。方法:选择体重250～350 g的健康雄性SD大鼠72只,随机分为3组,缺血再灌注组、二甲基乙二酰基甘氨酸(DMOG)组、假手术组,每组24只。缺血再灌注组:建立左肺缺血和再灌注动物模型:在肺充气状态下用无损伤动脉钳夹闭左侧肺门,阻断45 min后松开动脉夹,形成再灌注;DMOG组:在缺血和再灌注动物模型建立前24 h进行腹腔内注射DMOG 20μg/g;假手术组:仅行左侧开胸,不行缺血再灌注处理。缺血再灌注组与DMOG组于再灌注1 h、3 h、6 h、12h后颈动脉放血处死大鼠,假手术组于开胸后1 h、3 h、6 h、12 h颈动脉放血处死大鼠,取左肺上半部制成匀浆,硫代巴比妥酸比色法检测丙二醛含量、黄嘌呤氧化酶法检测超氧化物歧化酶活力,酶联免疫吸附法测定匀浆内白细胞介素-8含量,下半部通过免疫组化法行HIF-1α蛋白表达的观察与光镜下苏木素伊红(HE)染色后肺组织病理学改变的观察。结果:HIF-1α蛋白主要表达于肺泡上皮细胞的胞核或胞浆,与假手术组比较,缺血再灌注组、DMOG组各时间点HIF-1α蛋白的表达增强,DMOG组较缺血再灌注组各时间点HIF-1α蛋白的表达增强;缺血再灌注组、DMOG组在再灌注后3 h、6 h、12 h(缺血再灌注组12 h除外)较1 h HIF-1α蛋白的表达增强,差异有统计学意义(P<0.05或P<0.01)。与假手术组比较,缺血再灌注组、DMOG组各时间点白细胞介素-8(除DMOG组1 h外)、丙二醛含量均升高,而超氧化物歧化酶活力降低;DMOG组较缺血再灌注组各时间点白细胞介素-8、丙二醛含量均降低,而超氧化物歧化酶活力升高;缺血再灌注组、DMOG组在再灌注后3 h、6 h、12 h较1 h:白细胞介素-8含量(除DMOG组3 h外)升高,丙二醛含量(除缺血再灌注组、DMOG组12 h外)升高,超氧化物歧化酶活力降低,差异均有统计学意义(P<0.05或P<0.01)。光镜下DMOG组肺组织炎性渗出较缺血再灌注组减轻。结论:HIF-1α通过降低肺组织中丙二醛、白细胞介素-8含量,增强肺组织中超氧化物歧化酶活力,减轻肺内毛细血管充血及炎性细胞浸润,对大鼠肺缺血与再灌注损伤发挥保护作用。     

风湿性心脏瓣膜病慢性心房颤动患者白细胞介素-1β和肿瘤坏死因子-α蛋白表达的研究

目的 观察风湿性心脏瓣膜病慢性心房颤动(房颤)患右心耳白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)蛋白表达的改变。方法将48例接受心脏外科手术的风湿性瓣膜病患于术中获取的右心耳分为两组,其中窦性心律组27例,慢性房颤组21例,采用病理学检查评价心房组织炎症细胞浸润和纤维化,采用免疫组织化学检查评价IL-1β和TNF-α蛋白表达的变化。结果慢性房颤患心房组织有显的纤维化,而且其心房肌细胞IL-1β和TNF-α的表达强度也显大于窦性心律组。结论风湿性心脏瓣膜病慢性房颤患心房组织IL-1β和TNF-α蛋白的表达显增加。炎症反应可能是风湿性心脏瓣膜病慢性房颤患房颤发生和维持的机制之一。     

肿瘤坏死因子-α在大鼠脑缺血再灌注损伤中的作用机制

目的　探讨肿瘤坏死因 α(TNF α)在大鼠脑缺血再灌注损伤中的作用机制 ,尝试应用抗TNF α单抗进行缺血后的脑保护治疗。　方法　分别应用免疫组化方法和分光光度计比色法监测经大脑中动脉闭塞 2h后tor大鼠脑缺血再灌注不同时点TNF α的表达规律及髓过氧化物酶 (MPO)活性 ,并应用四氮唑红 (TTC)染色法测量再灌注 2 4h脑梗死灶体积。　结果　脑缺血再灌注后 2 4hTNF α表达和白细胞活性均达到高峰〔分别为 (3 7 86± 9 78)个和 (0 2 90± 0 0 71)U/ g湿组织 ;对照组分别为 (1 83± 1 41)个和 (0 0 16± 0 0 0 3 )U/ g湿组织〕 ,二者呈现同步变化。应用抗TNF α抗体可减少白细胞聚集〔治疗组为 (0 148± 0 0 3 3 )U/g湿组织〕和脑梗死灶体积〔手术组为 (15 8 7± 8 1)mm3,治疗组为 (86 3± 12 2 )mm3〕。　结论　TNF α通过炎性反应参与了缺血再灌注脑损伤 ,应用抗TNF α抗体可起到缺血性脑保护作用     

肿瘤坏死因子—α与脑缺血再灌注损伤

脑缺血再灌注后脑内肿瘤坏死因子－α（ＴＮＦ－α）的表达增加,在再增注损伤病理过程中ＴＮＦ－α具有损伤与修复双重作用。其作用机制与致炎、促凝血机制、血脑屏障破坏,诱导缺血耐受及神经生长因子生成等有关。     

肿瘤坏死因子—α在脑缺血再灌注损伤中的双刃剑作用

肿瘤坏死因子－α是一种具有广泛生物学活性的细胞因子，在脑缺血再灌注后表达增加。以往，人们注重的是其在脑缺血再灌注损伤中的细胞毒性，而近年来，随着研究的深入，已有一些研究证实，肿瘤坏死因子－α尚有神经保护和促进修复等作用，因此，肿瘤坏死因子－α在脑缺血再灌注损伤中具双刃剑作用。     

肿瘤坏死因子-α与脑缺血再灌注损伤

作为一种重要的炎性细胞因子,肿瘤坏死因子-α(tulllor necrosis factor-α,TNF-α)在脑缺血再灌注过程中起重要作用.探讨TNF-α,在脑缺血再灌注损伤过程中的动态变化、神经毒性作用机制以及拮抗TNF-α的治疗作用,将为脑血管病的治疗提供理论依据.     

TLR4 mediates LPS-induced HO-1 expression in mouse liver： Role of TNF-α and IL-lβ

AIM: Heme oxygenase (HO)-I catalyzes the conversion of heme to biliverdin, iron and carbon monoxide. HO-1 is induced by many stimuli including heme, Hb, heat stress,lipopolysaccharide (LP5) and cytokines. Previous studies demonstrated that LP5 induced HO-1 gene activation and HO-1 expression in liver. However, the mechanisms of LPS-induced HO-1 expression in liver remain unknown. The effect of toll-like receptor-4 (TLR4) on LPS-induced liver HO-1 expression and the role of TNF-α and IL-1β in this condition were determined.METHODS: HO-1 expression was determined by immunofluorescent staining and immunoblotting. Double immunofluorescent staining was performed to determine the cell type of HO-1 expression in liver.RESULTS: A low dose of LPS significantly increased HO-1 expression in the liver which was localized in Kupffer cells only. Furthermore, HO-1 expression was enhanced by three doses of LPS. HO-1 expression was significantly inhibited in the liver of TLR4 mutant mice. While the liver HO-1 expression in TNF KO mice was much lower than that in C57 mice following the same LPS treatment, IL-1β KO had a slight influence on liver HO-1 expression following LPS treatment.CONCLUSION: The preserfl: results confirm that macrophages are the major source of HO-1 in the liver induced by LPS.This study demonstrates that TLR4 plays a dominant role in mediating HO-1 expression following LPS. LPS-induced HO-1 expression is mainly mediated by endogenous TNF-α, but only partially by endogenous IL-1β.     

TLR4对肝脏缺血再灌注损伤大鼠肝组织Th17细胞相关细胞因子表达的影响

目的 观察Toll样受体4（TLR4）对肝脏缺血再灌注大鼠肝组织Th17细胞相关因子[IL-17A、IL-23、孤核儿相关受体（RORrt）]表达的影响,并探讨其可能机制.方法 将40只健康雄性SD大鼠分为4组各10只,假手术组只分离同侧的颈总动脉和股动静脉,并进行相应插管注入相同剂量的肝素;模型组、抗TLR4组、同型抗体组均制备创伤失血性休克再灌注肝损伤模型,抗TLR4组、同型抗体组分别于再灌注前10 min经股静脉注入TLR4抗体50μg、同型对照抗体50 μg,整个过程用微量注射器缓慢注射.于12 h后处死取材.采用Western blot法检测各组肝组织TLR4蛋白,ELISA检测IL-17A、IL-23蛋白,观察IL-17A、IL-23、RORrt mRNA,并对模型组、抗TLR4组、同型抗体组IL-17A、IL-23、RORrt进行相关性分析.结果 模型组TLR4蛋白表达较假手术组相比明显升高（P＜0.01）,应用抗TLR4抗体后表达明显减少（P＜0.01）,而同型抗体组较模型组无明显差异（P＞0.05）.与假手术组比较,模型组、抗TLR4组、同型抗体组IL-17A、IL-23蛋白及mRNA表达升高,RORrt mRNA表达升高;与模型组比较,抗TLR4组IL-17A、IL-23蛋白及mRNA表达降低,RORrt mRNA表达降低（P＜0.05或0.01）.IL-23水平与IL-17A水平呈正相关,IL-23 mRNA水平与RORrt mRNA呈正相关.结论 中和TLR4可减少IL-23、IL-17A、RORrt的表达;机制可能是中和阻断TLR4可通过降低IL-23的表达,影响Th17特异性转录因子RORrt,进而降低肝脏缺血再灌注损伤大鼠肝组织Th17/IL-17A的表达.     

Neutrophil-mediated liver injury during hepatic ischemia-reperfusion in rats

BACKGROUND: Neutrophil plays an important role in hepatic ischemia-reperfusion injury. We investigated neutrophil infiltration in liver tissue, Kupffer cells' role in neutrophil accumulation, and apoptosis and regeneration of hepatocytes in liver ischemia-reperfusion injury. METHODS: Vascular microclamps were placed across the pedicles of the median and left lateral lobes for 90 minutes after 30% hepatectomy with the resection of caudate, right lateral and quadrate lobes and papillary process. Gadolinium chloride (GdCl3) was used to destroy Kupffer cells. Neutrophil activity was inhibited with Urge-8, a monoclonal antibody against neutrophil produced in our laboratory. GdCl3 (10 mg/kg) and Urge-8 (50 mg/kg) were given intravenously in respective groups. Ischemia control, GdCl3 and Urge-8 groups were compared. RESULTS: Following hepatic reperfusion, serum interleukin-8 (IL-8) levels and hepatic neutrophil counts peaked at 3 hours, and peak concentrations of alanine aminotransferase (ALT) occurred at 6 hours. Animals of the control group showed increases in neutrophil infiltration in liver tissue, liver enzyme levels, and apoptosis index of hepatocytes and decreases in overall survival rate and proliferating cell nuclear antigen (PCNA) expression of hepatocytes. The survival rates and PCNA proportion of hepatocytes were higher and the levels of hepatic neutrophil infiltration, liver enzymes, and hepatocyte apoptosis after reperfusion were lower in the GdCl3 and Urge-8 groups than those in the ischemia control group. CONCLUSIONS: Blockades of Kupffer cells' activity and neutrophil infiltration by GdCl3 and Urge-8 eliminate neutrophil-mediated hepatic injury and enhance subsequent hepatic regeneration during liver ischemiareperfusion.     

大鼠肝缺血再灌注损伤肝核因子-κB、髓过氧化物酶、组织因子与凝血功能紊乱

肝缺血再灌注（IR）过程中，白细胞和细胞因子等不仅参与了肝组织炎症反应，而且极易引起凝血功能的紊乱。组织因子（TF）是一种跨膜糖蛋白，是凝血过程的启动因子，在炎症介质和内毒素等的刺激下，血管内皮细胞和单核细胞及巨噬细胞可表达TF。研究证实，核因子-κB（NF-κB）在肝IR损伤中发挥着重要作用。现旨在探讨肝I／R损伤中肝组织的炎性浸润与所引起的血液凝血功能失常之间的关系。     

三七总皂甙对大鼠肝缺血再灌注损伤的保护

三七总皂甙对大鼠肝缺血再灌注损伤的保护梁力建１何强２吕明德１彭宝冈１黄洁夫Ｔｈｅｐｒｏｔｅｃｔｉｖｅｅｆｅｃｔｓｏｆｐａｎａｘｎｏｔｏｇｉｎｓｅｎｇｏｎｌｉｖｅｒｉｓｃｈｅｍｉａ－ｒｅｐｅｒｆｕｓｉｏｎｉｎｊｕｒｙｉｎｒａｔｓＬＩＡＮＧＬｉ＿Ｊｉ...     

N-乙酰半胱氨酸对大鼠肺缺血再灌注损伤的保护作用

目的探讨N-乙酰半胱氨酸(NAC)对大鼠在体肺缺血再灌注(I/R)损伤的保护作用。方法建立大鼠在体肺缺血再灌注模型,将30只SD大鼠随机分成假手术对照组,缺血再灌注组(I/R组)和N-乙酰半胱氨酸组(NAC组),NAC组缺血前1 h给予腹腔注射N-乙酰半胱氨酸200 mg/kg。再灌注2 h后摘取左肺,分别对各组进行以下检测:肺湿/干比(W/D)、超氧化物歧化酶(SOD)活力、髓过氧化物酶(MPO)活性、丙二醛(MDA)含量并进行病理学检查及肺组织损伤定量评价(IQA)。结果I/R组肺W/D和IQA显著高于假手术组(P0.01),NAC组上述指标明显降低(P0.01)。病理学结果显示三组动物肺组织结构基本正常,假手术组无充血;与NAC组比较,I/R组肺组织充血明显、白细胞浸润更严重及肺间质高度淤血水肿。I/R组MDA含量和MPO活性较假手术组明显升高(P0.01),SOD活性显著下降(P0.01)。NAC能明显减少MDA含量和降低MPO活性,提高SOD活性(P0.01)。结论N-乙酰半胱氨酸对肺缺血再灌注损伤具有保护作用,可能与其抗氧化作用和抑制中性粒细胞激活有关。