纤维蛋白沉积对老年急性肺损伤大鼠肺组织内皮细胞功能的影响

目的 探讨纤维蛋白沉积对不同鼠龄大鼠肺组织血管内皮细胞功能的影响.方法 青年及老年大鼠均随机分为4组,对照组（NC）腹腔注射生理盐水;脂多糖组（L）腹腔注射脂多糖;氨甲环酸组（LT）注射脂多糖加氨甲环酸;尿激酶组（LTU）注射脂多糖及氨甲环酸加尿激酶.应用免疫组化分析纤维蛋白沉积;应用Western blot和Northern blot分别检测肺组织血栓调节蛋白（TM）、细胞间黏附分子-1（ICAM-1）、纤溶酶原激活剂抑制物-1（PAI-1）的表达.结果 （1）正常青年和老年大鼠肺组织内均无纤维蛋白沉积,脂多糖刺激后纤维蛋白沉积增多,加用氨甲环酸纤维蛋白沉积更加明显（P＜0.01）,应用尿激酶后纤维蛋白沉积明显减轻（P＜0.05）;老年大鼠L组、LT组和LTU组均比相同干预组的青年大鼠肺组织纤维蛋白沉积明显（P＜0.05）.（2）青年和老年大鼠给与脂多糖刺激后,ICAM-1和PAI-1表达明显上调（P＜0.05）,TM表达显著下降（P＜0.01）;加用氨甲环酸后ICAM-1和PAI-1表达上调更加明显（P＜0.01）,TM几乎无表达;给予尿激酶处理后,ICAM-1和PAI-1表达与LT组比较明显下调（P＜0.05）,TM表达增多（P＜0.05）;老年大鼠L组、LT组和LTU组Ⅰ CAM-1和PAI-1的表达均较青年大鼠相同干预组显著增多（P＜0.05）,TM表达则显著减少.（3）经纤维蛋白沉积量校正后,老年大鼠肺组织ICAM-1和PAI-1的表达量仍显著高于青年大鼠（P＜0.05）.结论 增龄能够促进脂多糖诱导的急性肺损伤大鼠肺组织纤维蛋白沉积,增强纤维蛋白的内皮细胞损伤作用.     

增龄对纤维蛋白急性肺损伤大鼠肺组织单核巨噬细胞趋化因子-1表达的影响

目的研究增龄对纤维蛋白诱导急性肺损伤大鼠肺组织单核巨噬细胞趋化因子-1 (MCP-1)表达的影响,探讨老年大鼠对炎性刺激易感的可能机制。方法青年及老年大鼠均根据腹腔注射不同的药物随机分为4组:对照组、脂多糖组、氨甲环酸组、肝素组。应用免疫组化分析纤维蛋白沉积;应用Western blot和Northern blot方法分别检测肺组织MCP-1蛋白质和基因表达。结果(1)青年和老年对照组大鼠肺组织内均无纤维蛋白沉积,老年大鼠脂多糖组、氨甲环酸组和肝素组均比青年大鼠相同干预组纤维蛋白沉积明显,差别均有统计学意义(18．5％±3．1％对12．3％±2．1％,32．5％±5．4％对24．1％±4．5％和20．8％±3．6％对12．6％±1．8％,均为P<0．05);(2)青年和老年对照组大鼠肺组织内几乎无MCP-1表达,老年大鼠脂多糖组、氨甲环酸组和肝素组的MCP-1表达均较青年大鼠相同干预组上调,差异有统计学意义(0．40±0．02对0．20±0．03,0．60±0．05对0．25±0．04和0．30±0．01对0．20±0．04,均为P<0．05);(3)经纤维蛋白沉积增加量校正后,老年氨甲环酸组大鼠肺组织MCP-1的表达量仍显著高于青年大鼠氨甲环酸组(P<0．05)。结论纤维蛋白可上调急性肺损伤肺组织MCP-1基因及蛋白质表达,促进肺组织炎症反应;增龄能够促进肺组织纤维蛋白沉积,增强其上调肺组织MCP-1表达的作用。     

Early effects of fibrin sealant on colonic anastomosis in rats: an experimental and case-control study

Background Leakage from colonic anastomoses leads to mortality and morbidity. Fibrin adhesives can be used to increase the strength of the anastomosis. In this study, we evaluated the early effects of fibrin sealant and hyaluronic acid-carboxymethylcellulose on colonic anastomosis in rats. Methods Anastomoses were made in the descending colon of 38 female Wistar-Albino rats, in three groups: control group (n=12), group 1 treated with hyaluronic acid-carboxymethylcellulose (n=16), and group 2 treated with fibrin sealant (n=10). After 72 hours, adhesion scores, bursting pressure, rupture strength and histopathologic healing scores were evaluated. Results Due to postoperative mortality, we evaluated 10, 10 and 9 rats in the control group and in groups 1 and 2, respectively. Of these, we excluded 4, 5 and 4 rats that had macroperforations at autopsy. In the remaining rats, bursting pressure (123.2±14.8 mmHg) and rupture strength (400±16 mg) in the fibrin sealant group were significantly greater than in the two other groups (Control: 68.0±10.6 p=0.006 and 325±52 p=0.009; Group 1: 74.0±9.8 p=0.03, 330±27 p=0.016). However, we did not observe any significant difference between adhesion scores (2.5±0.6, 2.0±0.7, 2.0±0.7, p=0.343). Conclusions In this experimental study, fibrin sealant increased bursting pressure and rupture strength of colonic anastomoses while hyaluronic acid-carboxymethylcellulose had no effetcs in rats, but both of them showed no effect on adhesion scores. In order to use fibrin sealant to decrease the rate of early leakages from colonic anastomoses, further studies have to be performed.     

双歧杆菌抑制小鼠实验性结肠炎肠道炎症反应的研究

目的观察补充益生菌对葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎的影响,探讨益生菌对结肠炎的治疗作用和可能机制。方法将小鼠分为4组:正常对照组、DSS组、柳氮磺胺吡啶(SASP)组和双歧杆菌组。建立小鼠急性DSS结肠炎模型。SASP组在饮DSS水同时予SASP灌胃。双歧杆菌组在实验开始前7d予双歧杆菌灌胃至实验结束。每天观察各组疾病活动指数(DAI),并在实验结束后检测各组小鼠炎症肠段肿瘤坏死因子(TNF)α、核因子(NF)κBP65、髓过氧化物酶(MPO)等的表达。结果自实验第4天开始,SASP组和双歧杆菌组DAI明显低于DSS组。实验结束时,SASP组和双歧杆菌组炎症肠段TNFα[(10.01±1.11)和(10.45±0.98)pg·mg-1·ml-1]、MPO[(3.21±0.20)和(3.24±0.16)U/g组织]等的水平较DSS组[(13.35±1.01)pg·mg-1·ml-1和(3.63±0.23)U/g组织]均明显降低,NFκBP65阳性的炎症细胞数减少。结论给小鼠预服双歧杆菌能有效抑制炎症肠段炎症细胞NFκB的活化及炎性细胞因子的分泌,减轻肠道炎症反应。     

Effects of primary suture and fibrin sealant on hemostasis and liver regeneration in an experimental liver injury

AIM： To investigate the effects of fibrin sealant on hemostasis and liver regeneration and intra-abdominal adhesions in an experimental liver injury. METHODS： Thirty-six Wistar rats were randomly divided into primary suture group （n = 15）, fibrin sealant group （n = 15） and control group （n = 6）. A wedge resection was performed on the left lobe of the liver. In primary suture group, liver was sutured using polypropylene material, while fibrin glue was administrated on the liver surface in fibrin sealant group. RESULTS： More intra-abdominal adhesions were observed in the primary suture group compared to the fibrin sealant group on 3rd （2.50 ± 0.5 vs 0.25 ± 0.5, P = 0.015）, 10^th （2.75 ± 0.5 vs 0.50 ± 0.6, P = 0.06） and 20^th （1.75 ± 0.5 vs 0.70 ± 0.5, P = 0.015） postoperative days. Histopathological scores were better in the fibrin sealant group in comparison with the primary suture group on 3rd （8.75 ± 0.5 vs 6.75 ± 0.5, P = 0.006）, 10th （7.50 ± 1.0 vs 5.5 ±0.6, P = 0.021） and 20th （6.40 ± 1.7 vs 3.20 ± 1.6, P = 0.025） postoperative days. CONCLUSION： Out data suggest that fibrin sealant is preferred over primary suture in appropriate cases including liver trauma since it causes less intra-abdominal adhesions while allowing shorter hemostasis time as assessed in experimental liver trauma.     

纤维蛋白原、纤维蛋白及其降解产物对共培养血管内皮细胞纤溶活性的影响

目的研究纤维蛋白原（Fg）、纤维蛋白（Fb）及其降解产物（FDP）对共培养体系中人脐静脉内皮细胞组织型纤溶酶原激活物和纤溶酶原激活物抑制剂表达的影响。方法应用Transwell膜建立人脐静脉内皮细胞-兔主动脉平滑肌细胞共培养体系，在不同浓度（0、0．5、1．5、3．0、4．5和6．0g／L）Fg、Fb和FDP干预24h后，分别检测该共培养体系中人脐静脉内皮细胞组织型纤溶酶原激活物和纤溶酶原激活物抑制剂mRNA水平（RT-PCR法）以及培养上清中组织型纤溶酶原激活物和纤溶酶原激活物抑制剂抗原含量（ELISA法）与活性（发色底物法）的变化情况。结果Fg对组织型纤溶酶原激活物的表达没有显著影响，较高浓度的Fg（3．0～4．5g／L）可明显促进纤溶酶原激活物抑制剂mRNA表达、抗原含量及活性升高，但过高浓度的Fg（6．0g／L）却抑制纤溶酶原激活物抑制剂的表达。3．0-4．5g／L的Fb对组织型纤溶酶原激活物mRNA和抗原含量都起上调作用，同时显著下调组织型纤溶酶原激活物活性。较高浓度的Fb（1．5-4．5g／L）则可明显上调纤溶酶原激活物抑制剂的表达，且在mRNA、蛋白和活性水平趋势基本一致。3．0-6．0g／L的FDP均可明显下调组织型纤溶酶原激活物mRNA、蛋白和活性水平，1．5-6．0mg／ml的FDP均可促进纤溶酶原激活物抑制剂的高表达。结论Fg、Fb和FDP可以通过影响组织型纤溶酶原激活物和纤溶酶原激活物抑制剂的表达，引起纤溶活性降低，参与动脉粥样硬化的发展进程。     

硒对氟致大鼠肾脏损伤保护作用的实验观察

目的 观察硒对氟致大鼠肾脏损伤的保护作用,探讨硒的最佳作用剂量及作用靶点.方法 断乳SD雄性大鼠80只,按体质量随机分8组,每组10只.对照组饮用自来水;染氟组饮用50 mg/L的氟化钠溶液;低、中、高硒组分别饮用0.375、0.750、1.500 mg/L的亚硒酸钠溶液;氟+低、中、高硒组分别饮用50 mg/L的氟化钠和0.375、0.750、1.500 mg/L的亚硒酸钠两两组合的溶液.染毒6个月后,测大鼠肾脏组织的氧化水平和核因子κB(NF-κB)的表达量.结果 染氟组大鼠体质量[(695.95±55.89)g]低于对照组[(782.69±56.12)g,P＜0.01],染氟组谷胱甘肽过氧化物酶(GSH-Px)活性[(55.86±5.09)U/mgprot]与对照组[(68.66±4.52)U/mgprot]比较,差异无统计学意义(P＞0.05),但有降低的趋势.染氟组大鼠总抗氧化能力(T-AOC)水平[(7.54±1.35)U/mgprot]低于对照组[(9.03±0.37)U/mgprot,P＜0.05],染氟组丙二醛(MDA)水平[(3.86±0.31)nmol/mgprot]高于对照组[(3.14±0.32)nmol/mgprot,P＜0.05].氟+高硒组GSH-Px活性[(74.99±8.41)U/mgprot]高于染氟组[(55.86±5.09)U/mgprot,P＜0.05],MDA水平[(3.17±0.20)nmol/mgprot]低于染氟组[(3.86±0.31)nmol/mgprot,P＜0.05].染氟组、高硒组和氟+低硒组的NF-κB的表达水平(0.360±0.015,0.367±0.007,0.376±0.006)高于对照组(0.312±0.022,P均＜0.05),氟+高硒组(0.312±0.005)低于染氟组(0.360±0.015,P＜0.05).结论 1.500 mg/L硒是本实验条件下硒对慢性氟中毒致大鼠肾脏损伤的最佳保护作用剂量,NF-κB可能是硒拮抗氟中毒的药物靶点.

Abstract：

Objective To explore the protective effect of selenium, an antioxidant, on fluoride-induced renal injury in rats and find out the optimal level of selenium against fluoride toxicity and its valid molecular target.Methods All 80 male weanling SD rats were randomly divided into 8 groups by body weight as follows: normal control group(drinking tap water), fluoride exposed group (drinking water containing 50 mg/L of NaF), low, middle,high selenium exposed groups(drinking water containing 0.375, 0.750, 1.500 mg/L of Na2SeO3) and low, middle,high Se-fluoride groups (drinking water containing both 50 mg/L NaF and three doses of Na2SeO3 as abovementioned, respectively). After 6 months, the rats were killed then the oxidation level and nuclear factor κB(NF-κB)expression level in kidney were measured. Results The weight of the fluoride exposed group[(695.95 ± 55.89 )g]was significantly deceased than the controls[(782.69 ± 56.12)g, P ＜ 0.01]. Glutathione peroxidase(GSH-Px)activity of fluoride exposed group[(55.86 ± 5.09)U/mgprot] was not significantly different but decreased. Tatal antioxidant capacity (T-AOC) activity in fluoride exposed group [(7.54 ± 1.35)U/mgprot] significantly decreased than the controls[(9.03 ± 0.37 )U/mgprot, P ＜ 0.05]. In addition, a significant increase of malondialdehyde ( MDA )in fluoride exposed group[(3.86 ± 0.31 )mnol/mgprot, P ＜ 0.05] was observed than the controls[(3.14 ± 0.32)nmol/mgprot, P ＜ 0.05]. GSH-Px activity of high Se-fluoride group[(74.99 ± 8.41 )U/mgprot] was significantly higher than the fluoride exposed group[(55.86 ± 5.09)U/mgprot, P ＜ 0.05] and its MDA level[(3.17 ± 0.20)nmol/mgprot] was lower than the fluoride exposed group[(3.86 ± 0.31 ) nmol/mgprot, P ＜ 0.05]. NF-κB expression levels of fluoride group, high selenium group and low Se-fluoride group(0.360 ± 0.015,0.367 ± 0.007,0.376 ± 0.006,respecyively) were obviously increased compared with the controls(0.312 ± 0.022, P ＜ 0.05); it was significantly lower in high Se-fluoride group(0.312 ± 0.005) than in fluoride exposed group(0.360 ± 0.015, P ＜ 0.05). Conclusions Na2SeO3 of 1.5 mg/L is the optimal dose against chronic fluorosis on kidney injury under this experimental condition.NF-κB is likely to be a target molecule of the selenium as an antagonist on fluorosis.     

Effects of alterations of glomerular fibrin deposition on renal inflammation in rats at different age stages

Recent data indicated that aging accelerated glomerular fibrin deposition induced by lipopolysaccharide (LPS) in mice. Our hypothesis was that aging may exacerbate glomerular inflammatory responses induced by glomerular fibrin deposition. Both young and aged rats with glomerular fibrin deposition induced by LPS were treated with tranexamic acid (TA) and TA plus urokinase (UK). Infiltrating inflammatory cells and expressions of monocyte chemoattractant protein 1, intercellular adhesion molecule 1, and vascular endothelial-cadherin were markedly upregulated in the LPS+TA group compared with the LPS group. Reduction of fibrin deposition in the LPS+TA+UK group was associated with downregulation of the above indices (p < .05), whereas the alteration of vascular endothelial-cadherin protein expression was negatively correlated with the fibrin deposition. There were also significant differences in increased expressions of monocyte chemoattractant protein 1 and intercellular adhesion molecule 1 between young and aged rats. These in vivo data demonstrated that fibrin deposition contributed to glomerular inflammatory responses, which could be exacerbated by aging.     

血管内皮生长因子及其受体随增龄在大鼠肾组织内表达的研究

目的　研究血管内皮细胞生长因子 (VEGF)及其受体Flt 1和Flk 1在大鼠肾组织内的表达及随增龄变化 ,探讨它们在肾脏衰老过程中的作用。　方法　应用 3、12、2 4月龄 (各 7只 )大鼠肾组织石蜡切片进行常规病理及免疫组织化学染色 ,定量分析肾组织内微血管变化及VEGF、Flt 1和Flk 1表达变化。应用逆转录聚合酶链反应技术 (RT PCR)检测肾组织内VEGF AmRNA的表达。　结果　 2 4月龄组与 3月龄组相比肾小球面积增大〔(15 6 35± 10 2 2 ) μm2 vs(72 0 5± 496 ) μm2 ,P <0 0 1〕 ,肾小球内毛细血管袢腔面积与肾小球面积百分比减少 (46 76 %± 4 91%vs 6 3 75 %±6 0 2 % ,P <0 0 1) ,肾小管周围毛细血管数量减少 (9 8± 2 6vs 14 7± 3 1,P <0 0 1) ;肾小球内VEGF阳性细胞数增多 (9 3± 2 4vs 6 4± 1 6 ,P <0 0 5 ) ;集合管中VEGF的表达则明显减少(9 35 %± 2 10 %vs 15 2 3%± 3 2 2 % ,P <0 0 5 ) ;Flk 1在肾小球血管袢上表达增加 (9 17%±2 0 2 %vs 1 0 3%± 0 35 % ,P <0 0 1) ,而Flt 1和Flk 1在肾小管上表达则明显减少 (7 6 4%±3 0 2 %vs 15 36 %± 2 5 4% ,2 48%± 0 86 %vs 9 0 1%± 2 6 3% ,P <0 0 1)。 2 4月龄组VEGF AmRNA较其他两组减少 (P <0 0 5 )。　结论　VEGF、Flk     

New strategies in the determination of fibrin and fibrin(ogen) derivatives by monoclonal antibodies

Summary Until recently only tests with a limited specificity were available for the assessment of the products of activated coagulation and/or fibrinolysis. Those assays were based on polyclonal antibodies, which crossreact with fibrinogen, and as a consequence they were performed on serum samples i.e. after removal of fibrinogen by clotting. Serum preparation, however, is a notorious source of artefactually high or low levels of fibrin(ogen) degradation products, and is not suitable for the determination of coagulation products. Recently, highly specific monoclonal antibodies (MoAb's) have been developed, the majority of which do not crossreact with fibrinogen. This has enabled new strategies to be developed, i.e. assays using these MoAb's on plasma samples. Furthermore, the new assays can discriminate between (individual) fibrin and fibrinogen degradation products, and coagulation products can be assessed in the same plasma samples.