Exogenous nitric oxide modulates the systemic inflammatory response and improves kidney function after risk-situation abdominal aortic surgery

OBJECTIVE: Renal impairment is a very frequent complication of aortic surgery requiring prolonged suprarenal clamping, especially if it is associated with previous hemorrhage. The aim of this study was to assess the beneficial effect of the administration of a nitric oxide (NO) donor on renal function through a modulation of the systemic inflammatory response in a model of abdominal aortic surgery. METHODS: Twenty-five minipigs were divided into five groups. Under anesthesia, the animals were subjected to suprarenal aortic-iliac clamping (for 30 minutes) and bypass with a Dacron-collagen prosthetic graft impregnated in rifampicin, with or without associated hemorrhage (40% of total blood volume). Prophylaxis with cefazolin was implemented. The five groups were (1) the sham group (only aortic dissection), (2) the clamping and bypass (C) group, (3) hemorrhage preclamping and bypass (H+C) group, (4) the same as group C but with the administration of the NO donor molsidomine (4 mg/kg intravenously) (C+NO group), (5) the same as the H+C group but with the administration of the NO donor molsidomine (4 mg/kg intravenously) (H+C+NO group). The following were determined: (1) kidney function (serum creatinine), (2) serum cytokines (tumor necrosis factor alpha [TNF-alpha] and interleukin-10 [IL-10]); (3) neutrophil infiltration (myeloperoxidase [MPO]) in the kidney, (4) oxygen free radicals (superoxide anion [SOA] and superoxide dismutase [SOD]) in the kidney, (5) serum nitrites, (6) soluble and kidney tissue cell adhesion molecule (soluble intercellular adhesion molecule-1 [sICAM-1], soluble vascular cell adhesion molecule-1 [sVCAM-1], intercellular adhesion molecule-1 [ICAM-1], and vascular cell adhesion molecule-1 [VCAM-1]), (7) inducible nitric oxide synthase (iNOS) in the kidney, and (8) nuclear factor-kappaB (NF-kappaB) in the kidney. Determinations were made during ischemia at 15 minutes post-reperfusion; at 24, 48, and 72 hours; and on day 7. RESULTS: The different insults used in the experimental model led to deterioration in kidney function and an increase in the systemic (and renal) inflammatory response at all levels investigated. Treatment with an NO donor, both with and without associated hemorrhage, reduced the inflammatory response at the systemic (TNF-alpha and IL-10) and kidney (MPO, SOA, and SOD) levels, normalizing kidney function. Likewise, exogenous administration of NO improved the excessive production of NO (nitrites) via iNOS. This was also reflected in a reduction in CAMs and of NF-kappaB expression. The hypotension induced by molsidomine was transitory and did not elicit hemodynamic repercussions. CONCLUSION: In our experimental model, prophylactic treatment with the NO donor molsidomine regulates the systemic inflammatory response and minimizes damage at the kidney level. Clinical Relevance The importance of this article resides in the fact that an experimental study that clarifies the effect of the donors of NO under circumstances as similar as possible to those of the human clinic, such as aortic surgery under hypovolemic shock (ruptured aortic aneurysm) have been little studied, most of these studies being performed in rodents without bypass. Using a model with one or two simultaneous insults (aortic clamping with/without previous hemorrhage) that is very similar to the human clinical situation (abdominal aortic rupture), we confirm the findings of previous work related to the beneficial effects of NO donors.     

Bacterial translocation as a source of Dacron-graft contamination in experimental aortic operation: the importance of controlling SIRS

BACKGROUND: Several experimental studies have shown the beneficial effects of nitric oxide (NO) in the modulation of the systemic inflammatory response syndrome (SIRS). Nitric oxide is involved in and affects almost all stages in the development of inflammation. We have attempted to ascertain whether the nitric oxide donor molsidomine prevents aortic graft contamination through control of the SIRS and a decrease in bacterial translocation (BT). METHODS: Twenty-four mini-pigs were divided into 4 groups. The animals were subjected to suprarenal aortic/iliac cross-clamping (for 30 minutes) and by-pass with a Dacron-collagen prosthetic graft impregnated in rifampicin. Groups: 1) sham (aortic dissection alone); 2) cross-clamping and bypass; 3) hemorrhage of 40% of total blood volume before cross-clamping and by-pass; and 4) the same as in group 3 but also including the administration of the NO donor molsidomine (4 mg/kg) 5 minutes before cross-clamping. Variables: 1) bacteriology of mesenteric lymph nodes (MLN), kidney, blood, and prosthesis; 2) serum TNF-alpha (ELISA); and 3) iNOS expression in kidney and liver (Western blot). RESULTS: Aortic cross-clamping with or without hemorrhage was associated with BT in 80% and 100% of the animals, respectively. About 86% of the bacteria isolated in the graft were also present in MLN. This contamination coincided with an increase in TNF-alpha and with a greater expression of iNOS. Molsidomine administration decreased TNF-alpha and iNOS, decreased BT (from 100% to 20% of the animals), and decreased graft contamination (from 83% to 20%). CONCLUSIONS: The present model induces high levels of BT and SIRS, both acted as sources of contamination for the implanted Dacron graft. Molsidomine administration decreased the presence of bacteria in the graft by controlling BT and modulating SIRS.     

Soluble tumor necrosis factor receptor prevents post-pump syndrome.

Post-pump syndrome is an acute lung injury following cardiopulmonary bypass (CPB) which is indistinguishable from the adult respiratory distress syndrome (ARDS). Tumor necrosis factor (TNF) is central to the inflammatory process and is capable of triggering the entire pathophysiologic response leading to ARDS. We hypothesized that treatment with a soluble TNF receptor-binding protein (TNFbp) would reduce the increase in serum TNF and prevent acute lung injury in our sequential insult model of ARDS following CPB. Anesthetized pigs were randomized to one of three groups: Control (n = 3), surgical preparation only; CPB + LPS (n = 6), femoral-femoral hypothermic bypass for 1 h followed by infusion of low dose Escherichia coli lipopolysaccharide (LPS; 1 microg/kg); and TNFbp + CPB + LPS (n = 4), pretreatment with intravenous TNFbp (2 mg/kg) followed immediately by both insults. CPB + LPS caused severe lung injury demonstrated by a significant fall in PaO2 and an increase in both intrapulmonary shunt and peak airway pressure as compared to all groups (P < 0.05). These changes were associated with a significant increase in plasma TNF level and pulmonary neutrophil sequestration. TNFbp significantly reduced plasma levels of TNF and prevented the lung injury typically observed with this ARDS model, but did not reduce pulmonary neutrophil sequestration. Thus, elevated serum TNF is not responsible for neutrophil sequestration but does play a role in neutrophil activation which causes lung injury. Prophylactic use of TNFbp in CPB patients may prevent neutrophil activation and reduce the incidence of post-pump ARDS.     

高密度脂蛋白对严重烫伤大鼠肺功能的保护作用

目的了解高密度脂蛋白（HDL）对严重烫伤大鼠肺功能的作用及其机制。方法将Wistar大鼠分为对照组（15只）、烫伤组（60只）和HDL组（60只）。对照组大鼠不作处理；其余2组均造成30％TBSAIII度烫伤，伤后常规补液，其中HDL组伤后立即经尾静脉注入HDL（80mg／kg）。检测2组烫伤大鼠伤后12、24、48、72h血清胞间黏附分子1（ICAM—1）及肿瘤坏死因子α（TNF—α）的含量，并检测其动脉血二氧化碳分压（PCO2）氧分压（PO2）；观察大鼠伤后各时相点肺组织病理学变化。另检测对照组上述指标。结果与对照组比较，烫伤组大鼠伤后各时相点ICAM—1、TNF—α的含量及PCO，均显著升高（P〈0．05或P〈0．01），PO，显著降低（P〈0．05）。HDL组上述指标与对照组比较，差异无统计学意义（P〉0．05）；但与烫伤组比较，各时相点ICAM—1、TNF—α含量均显著降低（P〈0，05或P〈0．01）。伤后48h，烫伤组ICAM-1、TNF—α分别为（3．42±0．25）μg／L、（4．04±0．28）ng／L，明显高于HDL组[（2．24±0．14）μg／L、（3．35±0．22）ng／L，P〈0．05或P〈0．01]。烫伤组大鼠伤后48h肺小血管周围炎性细胞浸润，肺内血管扩张出血，部分小血管管腔内红细胞淤滞凝集，形成“假血栓”；肺毛细血管内皮细胞连接松弛，内皮细胞水肿。HDL组肺内小血管周围炎性细胞浸润明显减轻；肺毛细血管内皮细胞连接较紧密。结论HDL对严重烫伤大鼠肺功能具有保护作用，可能与其抑制ICAM—1、TNF—α表达有关。     

Normal mesenteric lymph blunts the pulmonary inflammatory response to endotoxin

BACKGROUND: Mesenteric lymph may provide the mechanistic link between gut ischemia and acute lung injury after hemorrhagic shock (HS). Studies have focused on the toxic mediators that develop in the post-shock mesenteric lymph (PSML). However, a complementary possibility is that there is loss of protective mediators found in pre-shock normal mesenteric lymph (NML) after HS. We hypothesize that NML protects against inflammatory insults to the pulmonary endothelium and that this effect is lost in PSML. MATERIALS AND METHODS: Primary human pulmonary endothelial cells (HMVECs) were incubated with NML or PSML collected from rats subjected to HS and resuscitation and then stimulated with 20 ng/mL LPS. ICAM-1 surface expression was measured by flow cytometry. In subsequent experiments, lipoproteins were extracted from NML before incubation and LPS-induced ICAM-1 expression determined. RESULTS: Mean fluorescent intensity (MFI) of LPS-induced ICAM-1 in NML and PSML treated HMVECs were 10.1 +/- 2.3 versus 27.7 +/- 0.83, respectively (P < 0.05). This represented at 71% decrease in ICAM-1 expression by NML compared to ICAM-1 expression in LPS-induced controls (MFI: 34.6 +/- 6.9). Lipoprotein extraction from NML abolished this protective effect (MFI: 31.2 +/- 5.3 versus Control + LPS: 33.5 +/- 3.6, P > 0.05). Baseline ICAM-1 levels were not significantly different among control, NML, and PSML groups. CONCLUSION: Lipoproteins in NML contain anti-inflammatory properties that decrease ICAM-1 expression induced by LPS in pulmonary endothelium. Decreased protective lipoproteins after HS and resuscitation may contribute to the toxicity associated with PSML from the ischemic gut.     

Differential expression of inflammatory mediators in radiation-impaired wound healing

BACKGROUND: Radiation impairs healing, although the underlying mechanisms are not clearly defined. Normal healing requires a fine balance of promoting and inhibiting factors. We hypothesize that there may be a down-regulation of promoting factors (nitric oxide) and, in turn, an up-regulation of healing inhibiting factors (TNF-alpha and IFN-gamma) in the wound after radiation. MATERIAL AND METHODS: Groups of 10 rats were irradiated using single dose 12 or 24 Gy electron radiation at the dorsal skin. Control rats were sham-irradiated. On Day 5 a skin incision in the irradiated area was performed and polyvinyl alcohol sponges were inserted subcutaneously. Rats were sacrificed 10 days later to determine the wound-breaking strength and reparative collagen deposition. Nitrite and nitrate (index of NO synthesis), TNF-alpha, and IFN-gamma were measured within the wound fluid. Expression of the inducible NO-synthase (iNOS) was investigated by immunohistochemistry. Wound-derived fibroblasts were tested in vitro for NO and collagen synthesis. RESULTS: Irradiation significantly reduced wound collagen deposition and wound-breaking strength (P < 0.05). Impaired healing was reflected in diminished wound NO synthesis and iNOS expression (P < 0.01). TNF-alpha and IFN-gamma were increased in irradiated wounds (P < 0.05). Ex vivo, NO synthesis and collagen deposition by fibroblasts from irradiated rats were decreased (P < 0.01). In vitro irradiation of fibroblasts from nonirradiated rats decreased both NO and collagen production (P < 0.01). CONCLUSION: Radiation-impaired healing is reflected in an imbalance of promoting and inhibiting factors, leading to increased levels of TNF-alpha and IFN-gamma and decreased NO expression in the wound.     

G蛋白不同亚型在严重烫伤小鼠补体活化巨噬细胞分泌功能中的作用

目的观察严重烫伤后活化的补体对小鼠腹腔巨噬细胞(PM)分泌一氧化氮(NO)和肿瘤坏死因子(TNF)α的影响,探讨信号传递途径中不同G蛋白亚型的作用。方法血浆采集分组:补体血浆组,采用小鼠18%TBSAⅢ度烫伤模型;去补体血浆组,先在小鼠腹腔注射眼镜蛇毒素因子(CVF)去补体后再按上述标准烫伤。伤后6h分别收集两组小鼠全血制备血浆,用于培养正常小鼠的PM及经抑制型G蛋白(Gi)阻断剂百日咳毒素(PT)预处理的PM、经刺激型G蛋白(Gs)激活剂霍乱毒素(CT)预处理的PM,观察各组细胞培养上清液中NO及TNF-α含量的变化。结果补体血浆组培养上清液中的NO和TNF-α含量分别为(80±12)μmol/L和(46±6)%,明显高于去补体血浆组的(34±5)μmol/L和(26±5)%(P<0.01).PT预处理后补体血浆组PM产生的NO明显下降[(45±10)μmol/L,P<0.01],而TNF-α活性[(58±10)%]增加(P<0.05),CT预处理后补体血浆组PM产生的NO增加[(105±18)μmol/L,P<0.05],TNF-α的活性[(27±6)%]降低(P<0.01).结论严重烫伤后活化补体引起PM分泌NO和TNF-α增多这一现象,至少部分是通过G蛋白途径实现的。其中对PM生成NO的调控主要是通过Gi蛋白途径发挥作用,对PM分泌TNF-α的调控则以Gs蛋白信号通路为主。     

Matrix metalloproteinase inhibitor: differential effects on pulmonary neutrophil and monocyte sequestration following cardiopulmonary bypass

Acute respiratory distress syndrome (ARDS) following cardiopulmonary bypass (CPB), also known as "post-pump" or "post-perfusion syndrome" (PPS), results from sequential priming and activation of neutrophils. We hypothesized that chemically modified tetracycline (CMT-3) an inhibitor of neutrophil matrix metalloproteinase (MMP) and elastase, would prevent PPS. We performed histometric analysis of lung tissue from our porcine PPS model to correlate cellular sequestration and histologic injury with CMT-3 treatment. METHODS: Yorkshire pigs were randomized into five groups: Control (n = 3); CPB (n = 5); femoral-femoral bypass 1 hour; LPS (n = 7), Escherichia coli lipopolysaccharide (1 microgram/kg); CPB + LPS (n = 6); and CPB + LPS + CMT (n = 5), sequential insults and CMT-3. Protocol histometric analysis defined cellular and tissue components of lung injury. RESULTS: CMT-3 decreased neutrophil sequestration in the CPB + LPS + CMT-3 group (p < 0.0001 vs. CPB + LPS). There were no differences in monocytes between CPB + LPS and CPB + LPS + CMT treatment groups. CONCLUSIONS: CMT-3 attenuates neutrophil sequestration but has no effect on mononuclear sequestration in our PPS model. This finding supports current research on leukocyte chemokines and has important implications regarding mechanisms of CMT-3. Despite lack of monocyte response to CMT-3, PPS was prevented by inhibiting neutrophils alone; confirming the primary role of neutrophils in PPS.     

极化液对严重烫伤伴多器官功能障碍综合征大鼠炎性细胞因子水平及预后的影响

目的观察早期极化液治疗对烫伤伴多器官功能障碍综合征(MODS)大鼠炎性细胞因子水平及预后的影响,探讨其防治机制。方法将120只SD大鼠造成30%TBSAⅢ度烫伤,伤后2h腹腔注射内毒素,制作烫伤伴MODS大鼠模型,随机分为极化液组、葡萄糖组和盐水组,每组40只。观察3组大鼠伤后1~7d血浆中血糖、乳酸、肿瘤坏死因子α(TNF-α)、一氧化氮(NO)以及白细胞介素6(IL-6)含量的变化;并计算伤后7d内各组大鼠的死亡率。结果极化液组大鼠伤后1~7d血糖、乳酸、TNF-α、IL-6和NO含量均明显低于其他组(P<0.01),伤后6~7d其TNF-α、IL-6和NO含量达到最低值,分别为(2.37±0.54)μg/L、(0.28±0.17)μg/L及(29±9)μmol/L;葡萄糖组血糖、乳酸含量明显高于盐水组(P<0.01),而两组TNF-α、IL-6和NO含量相近(P>0.05).极化液组伤后7d内死亡率为20.0%,明显低于葡萄糖组与盐水组(分别为37.5%、47.5%,P<0.05).结论极化液通过改善烫伤引起的糖代谢障碍,降低机体炎性细胞因子水平,可作为防治烫伤伴MODS的辅助措施。     

Pentoxifylline attenuates stored blood-induced inflammation: A new perspective on an old problem

BACKGROUND: Blood transfusion is a risk factor for many inflammatory processes. Its supernatant fraction has been proven to activate neutrophils. We hypothesized that pentoxifylline (PTX) would attenuate stored blood-induced neutrophil activation and pro-inflammatory mediator production. METHODS: Whole blood was incubated with HBSS, LPS (100 microg/mL), leukoreduced PRBC supernatant + LPS, or supernatant + LPS + PTX (2 mmol/L). TNF-alpha levels were measured by ELISA. MMP-9 was evaluated with zymography. Neutrophil CD66b expression was determined by flow cytometry in blood treated with HBSS, fMLP (1 micromol/L), supernatant + fMLP, or supernatant + fMLP + PTX. RESULTS: TNF-alpha levels were elevated in both the LPS and supernatant + LPS groups (100%; P < 0.01 and 120%; P < 0.01, respectively). PTX administration resulted in a 106% decrease in TNF-alpha (P < 0.0001). MMP-9 levels were increased in all groups. Administration of PTX to the supernatant + LPS group generated a 33% decrease in MMP-9 levels, which was not statistically significant (P < 0.4). Upregulation of CD66b expression was seen in LPS and supernatant + LPS groups. Significant attenuation was seen with PTX (47%; P < 0.01). CONCLUSIONS: PTX downregulates CD66b and TNF-alpha expression in supernatant-induced whole blood. Because blood transfusion can contribute to inflammatory injury, the adjunctive use of PTX may have therapeutic potential.     

Angiotensin II effect on hydraulic permeability: interaction with endothelin-1, nitric oxide, and platelet activating factor

BACKGROUND: The purposes of this study were: 1) to examine the influence of endothelin (ET-1) release on the ability of angiotensin (Ang) II to modulate permeability, 2) to determine if the action of Ang II on microvascular permeability is dependent on nitric oxide (NO) release, and 3) to explore the effect of Ang II in microvessels activated with platelet activating factor (PAF). METHODS: Hydraulic permeability (L(p)) was measured using the modified Landis in vivo micro occlusion technique during perfusion with: 1) the ET-1 receptor antagonist PD145065 (50 microM), then PD145065 + Ang II, 2) Ang II (20 nm), a NO synthase inhibitor L-NAME (100 microM), then L-NAME + Ang II, and 3) after endothelial activation with 10 nM of PAF, then PAF + Ang II. RESULTS: 1) The ET-1 antagonist increased L(p) 2.5-fold, Ang II alone increased L(p) five-fold, while Ang II perfusion during ET-1 antagonism increased L(p) over 6-fold (P < 0.04); 2) L-NAME increased L(p) over 3-fold. Ang II perfusion during NO synthase inhibition had no effect compared to NO synthase inhibition alone (P = 0.9) while Ang II alone increased L(p) 5-fold (P < or = 0.01); 3) PAF + Ang II (L(p) = 2.74 +/- 0.12) was decreased versus PAF alone (L(p) = 4.66 +/- 0.25) (P < 0.02). CONCLUSIONS: Ang II does not increase hydraulic permeability via ET-1 release. Ang II may act via NO release to increase hydraulic permeability in the basal state. Finally, Ang II attenuates the increase in hydraulic permeability because of endothelial activation with platelet activating factor.     

Neutrophil depletion reduces myocardial apoptosis and attenuates NFkappaB activation/TNFalpha release after ischemia and reperfusion

BACKGROUND: This study tested the hypothesis that depletion of neutrophils (PMNs) reduces myocardial apoptosis via reducing oxidant generation and inhibiting NFkappaB-mediated signaling pathways after ischemia/reperfusion. METHODS: Anesthetized rats were randomly divided into one of four groups: Control: 30 min ischemia and 3 h of reperfusion; PMN depletion: anti-PMN serum was injected 6 h before ischemia; N-acetylcysteine (NAC): NAC was given twice before ischemia and at reperfusion. Sham: the ligature was placed without coronary occlusion. Apoptosis was detected by TUNEL staining and DNA fragmentation. PMN accumulation was studied by immunohistochemical staining. Levels of TNF-alpha, IL-6, and caspase-3 were detected by Elisa kits. Expression in NFkappaB, Bcl-2, and Bax was assessed by Western blotting analysis. RESULTS: Relative to Control, depletion of PMNs or NAC treatment reduced levels of plasma TNFalpha (567 +/- 130* and 231 +/- 72* versus 1994 +/- 447 pg/ml) and IL-6 (791 +/- 473* and 666 +/- 300* versus 3724 +/- 1233, pg/ml), accompanying a reduction in PMN accumulation (12 +/- 1* and 13 +/- 0.6* versus 20 +/- 1 mm2 myocardium) in ischemic myocardium. Both groups showed a reduction in expression of nuclear NFkappaB relative to Control (62 +/- 9* and 67 +/- 8* versus 124 +/- 16 arb.u), consistent with reduced NFkappaB binding activity. The number of apoptotic cells (%) in area at risk myocardium was comparably reduced in anti-PMN and NAC groups relative to Control (12 +/- 1* and 14 +/- 0.9* versus 20 +/- 1), consistent with reduced appearance of DNA ladders. Furthermore, activated caspase-3 was significantly reduced and Bcl-2 was increased relative to Control. No difference in all parameters measured was detected during the course of experiment in the Sham group. CONCLUSION: These data suggest that the oxidants generated from activated PMNs after ischemia/reperfusion trigger myocardial apoptosis, which is further supported by an anti-oxidant therapy with NAC, potentially mediated by enhanced NFkappaB-TNFalpha signaling pathway, activated caspase-3 and down-regulated Bcl-2. *P < 0.05 versus Control.     

Multiple sequential insults cause post-pump syndrome.

BACKGROUND: We hypothesize that post-pump syndrome (PPS) following cardiopulmonary bypass (CPB) can be caused by multiple minor insults and that the mechanism of PPS is a priming and subsequent activation of polymorphonuclear (PMN) leukocytes. In this study extensive pathophysiologic and morphometric assessment was undertaken in a porcine model of sequential insult PPS. METHODS: Pigs were anesthetized, placed on a ventilator, instrumented for measurements of hemodynamic function, and separated into five groups: (1) Control (n = 4)--surgery only, (2) CPB (n = 4)--placed on femoral-femoral hypothermic (28 degrees C) bypass for 1 h, (3) LPS (n = 6)--underwent sham CPB followed by infusion of low dose endotoxin [E. coli lipopolysaccharide (LPS-1 microg/kg)], (4) Heparin + protamine + LPS (HP + LPS, n = 4)--were heparinized without CPB for 1 h, following which protamine and LPS were infused and (5) CPB + LPS (n = 8)--subjected to both CPB and LPS. RESULTS: Only CPB + LPS resulted in acute respiratory distress typical of PPS as indicated by a significant decrease in PaO2 and increase in intrapulmonary shunt fraction (p<0.05). CPB + LPS significantly increased tissue density and the number of sequestered monocytes and PMNs (p<0.05) above all other groups. Alveolar macrophages (AM) increased equally in all groups receiving LPS. CONCLUSIONS: CPB primes the inflammatory system causing pulmonary PMN sequestration without lung injury. Exposure to an otherwise benign dose of endotoxin results in activation of the sequestered PMNs causing PPS. This study confirms that PPS can be caused by multiple minor insults.     

Ν-硝基-L-精氨酸甲酯对大鼠隐睾生殖细胞凋亡的保护作用

目的 :探讨一氧化氮合酶 (NOS)抑制剂N 硝基 L 精氨酸甲酯 (L NAME)对大鼠隐睾生殖细胞凋亡的保护作用。　方法 :用 2 2dSD雄性大鼠复制单侧隐睾模型。实验分假手术组、隐睾组、隐睾 +L NAME组 [术后腹腔注射L NAME ,10mg/(kg·d) ],每组大鼠各 10只。术后 7d ,用生物素 dUTP/酶标亲和素测定法检测睾丸生殖细胞凋亡 ,用硝酸还原酶法测定睾丸组织中N0含量 ,用化学比色法测定睾丸组织中NOS活性。　结果 :术后第 7d ,与假手术组睾丸相比 ,隐睾组睾丸发生凋亡的生殖细胞数显著增加 ,隐睾 +L NAME组睾丸发生凋亡的生殖细胞数比隐睾组显著减少 (P <0 .0 1) ,隐睾 +L NAME组睾丸组织中NO含量及NOS活性与隐睾组相比显著降低 (P<0 .0 1)。　结论 :隐睾组织中NO和NOS升高是隐睾生殖细胞凋亡增加的病理机制之一 ,L NAME通过抑制NOS活性、减少NO的产生来降低睾丸组织生殖细胞的凋亡发挥其保护作用。     

Inflammatory response on the pancreatic acinar cell injury.

Acute pancreatitis is an inflammatory disorder, and inflammation not only affects the pathogenesis but also the course of the disease. Acinar cell injury early in acute pancreatitis leads to a local inflammatory reaction; if marked this leads to a systemic inflammatory response syndrome (SIRS). An excessive SIRS leads to distant organ damage and multiple organ dysfunction syndrome (MODS). MODS associated with acute pancreatitis is the primary cause of morbidity and mortality in this condition. Recent studies by us and other investigators have established the critical role played by inflammatory mediators such as TNF-alpha, IL-1beta, IL-6, IL-8, CINC/GRO-alpha, MCP-1, PAF, IL-10, CD40L, C5a, ICAM-1, MIP1-alpha, RANTES, substance P, and hydrogen sulfide in acute pancreatitis and the resultant MODS. This review intends to present an overview of the inflammatory response that takes place following pancreatic acinar cell injury.     

Ulinastatin suppresses systemic inflammatory response following lung ischemia-reperfusion injury in rats

OBJECTIVE: We sought to investigate whether ulinastatin (urinary trypsin inhibitor) inhibited systemic inflammatory responses following lung ischemia-reperfusion (I/R) injury. MATERIALS AND METHODS: Establishing a steady left lung warm I/R model in rats, we randomly divided 32 animals into 4 groups: sham (n = 8); I/R (n = 8); low-dose ulinastatin (5000 U/kg pre-ischemia) + I/R (n = 8); and high-dose ulinastatin (10,000 U/kg pre-ischemia) + I/R (n = 8). Measured variables included plasma concentrations of tumor necrosis factor-alpha (TNF-alpha), as well as interleukin (IL)-6 and IL-8. RESULTS: The serum concentrations of TNF-alpha, IL-6, and IL-8 in the ulinastatin pretreated groups were markedly decreased compared with those of the I/R group (P < .05). The levels of TNF-alpha, IL-6, and IL-8 were lower in the high-dose ulinastatin group compared with the low-dose ulinastatin group (P < .05). CONCLUSION: Ulinastatin produced dose-dependent attenuation of the systemic inflammatory response of rats following lung I/R injury.     

核转录因子κB decoy寡核苷酸对移植静脉内膜增生的影响

目的 研究核转录因子κB(NFKB)decoy寡核苷酸(decoy ODNs)对移植静脉内膜增生的抑制作用,探讨其作用机制。方法 Wistar大鼠72只,建立自体静脉移植模型,术后随机分为：对照组,NFκB decoyODNs 50μg、200μg组,杂码decoyODNs 50μg、200μg组,lipofectin pluronic组等6个组,施加不同的处理方法,在移植后1、2周取材。组织形态学方法比较内膜增生程度,半定量RT-PCR方法检测细胞间黏附分子(ICAM)-1的mRNA表达,Western blot和免疫组化方法检测p65、ICAM-1的蛋白产物表达。结果 移植后1、2周内膜增生明显,局部应用50μg NFκB decoyODNs明显抑制内膜增生,抑制率为22％-31％;200μg组受抑制程度更为明显,抑制率达41％～53％,其他组内膜增生无明显变化。NFκB decoyODNs能够抑制：ICAM-1mRNA表达,p65和：ICAM-1的蛋白表达均显著低于各对照组,抑制率为30％-57％。结论 NFκB decoyODNs可显著抑制移植静脉的内膜增生,其作用可能是通过减少黏附分子的基因转录及蛋白产物表达而实现的。     

TNF-alpha neutralization decreases nuclear factor-kappaB activation and apoptosis during renal obstruction

BACKGROUND: Obstruction of the upper urinary tract is an important cause of progressive renal injury in children. While tumor necrosis factor-alpha (TNF-alpha) and nuclear factor kappaB (NFkappaB) have independently been implicated in the pathophysiology of this process, TNF-alpha's role in obstruction-induced NFkappaB activation has not previously been investigated. MATERIALS AND METHODS: To study this, male Sprague Dawley rats were subjected to 3 days of unilateral ureteral obstruction (UUO) versus sham operation. Twenty-four hours prior to surgery and 2 days after, rats received either a vehicle or a pegylated form of soluble TNF receptor type 1 (PEG-sTNFR1). The kidneys were harvested 3 days postoperatively, and tissue samples were analyzed for TNF-alpha expression (ELISA), NFkappaB activation (EMSA, immunohistochemistry), IkappaB degradation (Western blot), angiotensinogen expression (Western blot), and apoptosis (TUNEL). RESULTS: Renal cortical TNF-alpha levels, NFkappaB activation, IkappaB degradation, angiotensinogen expression, and apoptotic cell death were significantly increased in response to obstruction. In contrast, TNF-alpha neutralization significantly reduced obstruction-induced TNF-alpha production, NFkappaB activation, IkappaB degradation, angiotensinogen expression, and renal tubular cell apoptosis. CONCLUSION: TNF-alpha's potent pro-inflammatory and cytotoxic effect during renal obstruction is directed through NFkappaB activation via increased IkappaB-alpha phosphorylation. As the role of TNF-alpha and NFkappaB in renal obstruction are further defined, the development of therapeutic strategies that limit or prevent obstruction-induced renal injury may be realized.     

Fructose induces the inflammatory molecule ICAM-1 in endothelial cells

Epidemiologic studies have linked fructose intake with the metabolic syndrome, and it was recently reported that fructose induces an inflammatory response in the rat kidney. Here, we examined whether fructose directly stimulates endothelial inflammatory processes by upregulating the inflammatory molecule intercellular adhesion molecule-1 (ICAM-1). When human aortic endothelial cells were stimulated with physiologic concentrations of fructose, ICAM-1 mRNA and protein expression increased in a time- and dosage-dependent manner, which was independent of NF-kappaB activation. Fructose reduced endothelial nitric oxide (NO) levels and caused a transient reduction in endothelial NO synthase expression. The administration of an NO donor inhibited fructose-induced ICAM-1 expression, whereas blocking NO synthase enhanced it, suggesting that NO inhibits endothelial ICAM-1 expression. Furthermore, fructose resulted in decreased intracellular ATP; administration of exogenous ATP blocked fructose-induced ICAM-1 expression and increased NO levels. Consistent with the in vitro studies, dietary intake of fructose at physiologic dosages increased both serum ICAM-1 concentration and endothelial ICAM-1 expression in the rat kidney. These data suggest that fructose induces inflammatory changes in vascular cells at physiologic concentrations.