Effect of Ligustrazine on liver injury after burn trauma

This study was designed to investigate the effect of Ligustrazine on burn-induced liver injury as well as the activation of nuclear factor kappaB (NF-kappaB) in severely burned rats. Sprague-Dawley rats were divided into three groups: (1) sham group, rats who underwent sham burn; (2) control group, rats given third-degree burns over 30% total body surface area (TBSA) and lactated Ringer solution for resuscitation; (3) Ligustrazine group, rats given burn and lactated Ringer's solution with Ligustrazine inside for resuscitation. Liver injury was assessed at 24 h post-burn by serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), as well as liver wet/dry weight ratio. Liver myeloperoxidase (MPO) activity was also analyzed. Hepatic NF-kappaB activity was examined by electrophoretic mobility shift assay (EMSA). Burn results in hepatic dysfunction and increased hepatic NF-kappaB activity, elevated liver wet/dry ratio and hepatic MPO activity. Ligustrazine inhibited these changes and alleviated burn-mediated hepatic dysfunction. The data indicated that Ligustrazine has a protective effect on burn-induced liver injury and possible mechanism may be attributed to its inhibitory action on the activation of NF-kappaB following burn trauma.     

The role of NO in macrophage dysfunction at early stage after burn injury

AIM: To explore the role of nitric oxide (NO) in macrophage dysfunction at early stage after burn injury. METHOD: Peritoneal macrophages were isolated and cultured from early stage burnt mice. NO production and inducible NO synthase (iNOS) expression in the macrophages were checked by the Greiss method and real-time PCR (TaqMan), respectively. l-Arginine, the substrate of NO producing, or N-monomethyl-l-arginine (l-NMMA), a competing blocker of NOS was administered to the culture, the changes of NO, TNF-alpha and PGE2 productions were measured, additionally the changes of the iNOS, TNF-alpha and COX-2 expression were assayed by real-time PCR. After that, the effects of l-arginine and l-NMMA were determined on burnt macrophage influencing the proliferation of normal splenic lymphocytes. RESULT: A large amount of NO was produced by macrophages from post burn hour 6 (6PBH) with a high level of iNOS expression. l-Arginine could increase NO production in a dosage-dependent manner, while l-NMMA attenuated NO production, but neither could affect iNOS expression. Moreover, l-arginine enhanced productions of both the latter produced TNF-alpha and PGE2 from burnt macrophages, and the expressions of TNF-alpha and COX-2 were improved significantly, while l-NMMA did reverse ways. It was found that macrophages from post burn hour 24 mice could inhibit Con A-stimulated normal splenic lymphocytes dramatically, l-NMMA could decrease this function significantly, but l-arginine could not influence the suppression. CONCLUSION: Our experiment indicated NO derived from burnt macrophage played a vital role in macrophage producing excessive TNF-alpha and PGE2, and suppressing lymphocyte function at early stage after burn injury.     

Major injury induces increased production of interleukin-10 in human granulocyte fractions

Patients with severe trauma or polytrauma frequently acquire alterations in immune functions which are correlated to dysbalanced cytokine synthesis. In these settings the role of polymorphonuclear neutrophil granulocytes (PMN) as cytokine-producing cells is less well characterized. The immunosuppressive role of interleukin (IL)-10 is well known, and increased systemic IL-10 levels are related to the severity of injury and to posttraumatic complications. We determined concentrations of IL-10 in culture supernatants of 30 individual PMN fractions isolated from 18 severely traumatized patients (15 polytraumata, Injury Severity Score: 18–41, 3 severely burned patients) admitted to intensive care units. IL-10 was analyzed by ELISA (R&D Systems, Wiesbaden, Germany). PMN were isolated from EDTA-anticoagulated peripheral blood employing a one-step procedure based on a discontinuous double Ficoll gradient. The cells [1×10⁶/ml RPMI 1640 supplemented with 10% fetal calf serum and 25 mM N-(2-hydroxyethyl)-piperazine-N′-(2-ethanesulfonic acid] were stimulated with 0.05% heat-killed Staphylococcus aureus (Pansorbin, Calbiochem-Novabiochem, Bad Soden, Germany) for 24 h using cell culture conditions. Our results show that PMN fractions of traumatized patients produce significantly (P<0.008) higher amounts of IL-10 (354± 95 pg/ml, n = 30) than normal healthy donor cells (125± 95 pg/ml, n = 7). IL-10 release from PMN fractions exceeded the release from isolated patients' peripheral blood mononuclear cells induced by similar stimulation or by stimulation with toxic shock syndrome toxin-1 (10 ng) and concanavalin A (2 μg). Our results provide evidence that PMN fractions play an active role in the development of posttraumatic immunosuppression by autocrine or paracrine mechanisms, for example, by suppressing one's own antimicrobial activities or determining the development of T-cell responses via their ability to release IL-10. Received: 9 March 1998; in revised from: 3 August 1998 / Accepted: 1 September 1998     

The effect of severe burn injury on proinflammatory cytokines and leukocyte behavior: its modulation with granulocyte colony-stimulating factor.

Severe injury causes immunosuppression. The main contributors are impaired leukocyte function and a cytokine dysbalance. GCSF increases PMN count, function and modulates the inflammatory response. However GCSF may overactivate leukocytes. The purpose of this study is to investigate whether GCSF is able to restore immune competence after severe injury. Lewis rats were divided into three groups: 30% TBSA burn + vehicle; 30% TBSA burn + GCSF (150 microg rhGCSF); Control. Blood samples were taken for total white cell count, PMNs, TNFalpha and IFNgamma. Leukocyte rolling and sticking were measured in the cremaster muscle microcirculation. Leukocyte diapedesis was investigated by lavage of the abdominal cavity and the lungs. Total white cell and PMN counts in the burn + GCSF group were significantly higher (P<0.001) than in burn+vehicle animals. Leukocyte adherence and diapedesis were not elevated in the burn + GCSF group as compared to the burn + vehicle group. TNFalpha (P<0.05) and IFNgamma (P<0.001) levels were significantly increased in the burn + vehicle animals compared to the burn + GCSF animals. GCSF modifies the immune system, as shown by an increase in white cell and PMN counts and by balancing the overall immune response from proinflammatory to normal, as shown by decreased TNFalpha and IFNgamma levels. GCSF does not overactivate PMNs.     

巨噬细胞及粒巨细胞-集落刺激因子对大鼠腹壁皮瓣成活的影响

目的 研究巨噬细胞及粒巨细胞-集落刺激因子(GM-CSF)对大鼠腹壁动脉穿支(deep epigastric perforator,DEP)皮瓣模型的影响.方法 建立SD大鼠DEP皮瓣模型,分别给予大鼠GM-CSF(Ⅰ组)、腹腔巨噬细胞(Ⅱ组)、GM-CSF联合巨噬细胞(Ⅲ组)及乍理盐水(Ⅳ组).术后第7天取皮瓣检测成活而积、组织学观察、微血管密度(MVD)、皮瓣内胶原含量.结果 皮瓣成活率Ⅰ组(53.08%±8.76%)和Ⅱ组(47.95%±4.92%)间无差异,均高于Ⅳ组(43.28%±5.27%)而低于Ⅲ组(61.68%±6.60%),P<0.05.皮瓣MVD Ⅰ组(24.82±4.18)和Ⅱ组(24.30±3.02)间差异无统计学意义,均显著高于Ⅳ组(21.37±2.65),低于Ⅲ组(29.82±4.74).胶原含量Ⅰ组(17.25%±2.85%)高于Ⅳ组(14.41%±2.89%),P<0.05.Ⅱ组(12.69%±3.55%)稍低于Ⅳ组.Ⅲ组(20.31%±3.01%)较Ⅰ组显著增高,P<0.05.结论 重组大鼠GM-CSF和巨噬细胞均能够促进大鼠DEP皮瓣成活,联合应用可发挥协同作用促进皮瓣存活、血管生成以及胶原沉积.     

NO在小鼠烧伤后腹腔巨噬细胞功能变化中的作用

目的探讨ＮＯ在烧伤后小鼠巨噬细胞（ＭΦ）功能变化中的作用。方法通过ＮＭＭＡ等的应用,观察体外培养烧伤后腹腔ＭΦ（ＰＭΦ）产生ＮＯ、ＰＧＥ２和ＴＮＦα的变化,以及对烧伤后ＰＭΦ抑制脾淋巴细胞（ＳＬ）增殖的影响。结果烧伤早期ＭΦ就能产生大量ＮＯ,ＮＯ能促进烧伤后ＰＭΦ产生ＰＧＥ２及ＴＮＦα,且烧伤后２４小时（２４ＰＢＨ）ＰＭΦ可通过产生的ＮＯ显著抑制ＣｏｎＡ刺激的ＳＬ增殖。结论ＮＯ在烧伤后ＭΦ功能变化乃至机体的免疫功能紊乱中起着重要作用。     

Effect of burn injury on corticosteroid-binding globulin levels in plasma and wound fluid

Corticosteroids exert inhibitory effects on wound healing. They circulate, largely bound to corticosteroid-binding globulin, and the plasma concentrations of this protein determine their bioavailability. The amount of corticosteroid-binding globulin in wounds and the related effects of burn injury are not known. We have therefore measured corticosteroid-binding globulin in serum and wound fluid obtained from subcutaneously implanted sponges, retrieved 1, 3, and 10 days after insertion in rats. The effect of burning was studied by comparing rats that had a small scald burn with sham-burned control rats. In serum, corticosteroid-binding globulin levels were lower in burned rats than in control animals: the difference was 22%, 28%, and 37% for days 1, 3, and 10, respectively ( p < 0.05 for each comparison), and values at day 1 were lower than at days 3 and 10 in control rats ( p < 0.05) but not in burned rats. In wound fluid, corticosteroid-binding globulin levels were lower in burned rats than in control animals: the difference was 23%, 24%, and 34% for days 1, 3, and 10, respectively ( p < 0.01 for all comparisons), and the values were significantly higher ( p < 0.05) at day 1 when compared with values at day 10 in both groups. We therefore conclude that a small burn injury has significant effects on levels of corticosteroid-binding globulin on serum and wound fluid corticosteroid-binding globulin. The decreased concentration of wound fluid corticosteroid-binding globulin at day 10 versus day 1, with a concomitant increase in serum corticosteroid-binding globulin, suggests an accelerated degradation of the protein within the wound; this phenomenon is exaggerated by the burn injury. This is supported by Western blot analysis, which revealed the appearance of a small polypeptide that reacts with an antiserum against rat corticosteroid-binding globulin in wound fluid at day 10.     

精氨酸在烧伤免疫营养治疗中的作用

精氨酸是一种含有两个碱性基团及氨基和胍基的氨基酸。正常成人体内有足够的精氨酸,可满足机体需要,但在人体发育不成熟和严重应激状态下,虽然体内能合成精氨酸,但其量不足,在这种情况下机体不能维持正氮平衡和正常的生理功能,需要从体外获得精氨酸。由此可见,精氨酸是人体的一     

Effect of burn injury on glucocorticoid receptor binding activity in rat muscle

Burn injury is associated with increased muscle proteolysis and up-regulated gene expression in the proteolytic pathway. Glucocorticoids (GCs) are the most important mediator of burn injury induced muscle cachexia. However, the understanding of the mechanisms of GCs action in response to burn injury remains elusive. It is well known that GC acts by binding its own receptor. Therefore, in the present study, we examined the influence of burn injury on the hormone binding activity of the glucocorticoid receptor (GR) in skeletal muscle. Burn injury resulted in increased hormone binding activity in extensor digitorum longus (EDL) and soleus muscles. Scatchard plots revealed that the increased GR hormone binding activity reflected increased numbers of hormone binding sites, whereas receptor affinity for GCs was unchanged. Western blot analysis showed that dissociation of GR/heat shock protein 90 heterocomplex or increase in GR protein may account for the effect of burn injury. The GR antagonist RU 38486 blocked the burn injury-induced increase in GR hormone binding activity, implicating a positive regulatory effect of GCs on GR binding activity under the present experimental conditions.     

Effect of lidocaine on reducing injury in a rat electrical burn model

Electrical injuries induce progressive tissue loss. We evaluated the effect of lidocaine on tissue necrosis after electrical burn injuries. Forty-two male Wistar albino rats (250-300 g) were divided into 3 groups [Group A (n=6), control group without an electrical burn injury; and Groups B (n=18) and C (n=18), electrical burn injury groups without and with lidocaine therapy, respectively]. Three separate analyses were performed at different time points on 6 of 18 rats from Groups B and C at each time point. Electrical burns were induced by applying 220 V AC between the left upper and right lower extremities for 10 seconds. Myeloperoxidase and malondialdehyde levels were measured in skin and muscle biopsy specimens after the first hour, fresh and dry weight differences in the amputated extremities were calculated after 24 hours, and live and necrotic tissue areas were measured at 7 days after burn injury. We found that lidocaine reduced edema, the number of neutrophils, and neutrophil damage in tissues. We conclude that lidocaine decreased the amount of necrotic tissue caused by electric injury.     

Acute burn injury

Almost 2 million people in the United States suffer from burns annually. A small percentage of these injuries are fatal, but all require some degree of medical attention. Burn injury is associated with anatomic, physiologic, endocrinologic, and immunologic alterations. These problems need to be identified and treated properly to prevent or minimize the extent of the damage. In recent years, advances in burn treatment have reduced morbidity and mortality and improved the quality of life for burn survivors. These advances have been made in the treatment of the acute injury, the quality of the initial resuscitation, the effectiveness of infection control, and the surgical decision making for improved short- and long-term outcomes.     

Recombinant human granulocyte colony-stimulating factor and Pseudomonas burn wound sepsis

Multiple immune defects have been demonstrated following thermal injury, including defective granulocyte production and function. Recombinant human granulocyte colony-stimulating factor (rhGCSF) is a regulator of the myelopoietic system. The effect of rhGCSF administration on survival and on the myelopoietic system in a murine model of Pseudomonas burn wound sepsis was investigated. Male BDF1 mice that underwent a 15% total body surface area burn injury and burn wound seeding with 1 x 10(8) Pseudomonas aeruginosa organisms demonstrated an improved mean survival time with the subcutaneous administration of 100 ng of rhGCSF twice a day. Mice that underwent a similar thermal injury and burn wound seeding with 3 x 10(7) P aeruginosa organisms demonstrated an augmented myelopoietic response through the administration of rhGCSF, as represented by significantly increased white blood cell count, neutrophil count, splenic weight, femoral marrow cellularity, and femoral marrow granulocyte-macrophage colony-forming cell count. Myelopoietic augmentation through rhGCSF administration may serve to decrease the morbidity of septic events following thermal injury.     

不同免疫调节剂对烫伤大鼠巨噬细胞功能的影响

目的探讨不同免疫调节剂对烫伤大鼠腹腔巨噬细胞（Ｍ）功能的影响。方法应用ＭｃＡｂＡＰＡＡＰ桥联酶标法、琼脂溶菌板法和ＭＴＴ比色法等技术,分别测定了各组烫伤大鼠不同时相腹腔Ｍ各种功能的变化。结果①烫伤后大鼠腹腔Ｍ表面Ｉａ抗原表达率降低,抗原提呈能力减弱,对念珠菌的噬菌率下降,溶菌酶活力减低,ＴＮＦ分泌增高,且伤后第１０天比第５天更明显。以上结果与正常组比较,差异有非常显著意义（Ｐ＜０．０１）。②经不同免疫调节剂治疗后,各组烫伤大鼠腹腔Ｍ各种功能明显改善,各项指标与未用药的对照组比较,差异有非常显著意义（Ｐ＜００１）。结论烫伤后早期腹腔注射特异性免疫核糖核酸（ｉＲＮＡ）可明显改善伤鼠的免疫功能。     

Some aspects of neutrophil granulocyte function in burn patients

The polymorphonuclear neutrophil leukocyte (PMN) functions and bacterial colonization of 18 adult burn patients were studied for at least three weeks following injury. Bacterial load was at a maximum in the second week after injury.PMN chemotaxis and random migration measured by the leading front technique was not changed. Phagocytosis measured as uptake rate of Ig coated latex particles was impaired in all patients examined. Bactericidal capacity against Staphylococcus aureus was impaired in 14 patients, at the most in the second week. Eight patients with poor and 1 patient with normal bactericidal capacity had positive blood cultures, mostly with Gram-positive organisms.The intracellular contents in PMNs of lactoferrin, myeloperoxidase and chymotrypsin-like cationic protein were markedly lowered in the majority of samples. This was well correlated to impairments in bactericidal capacity.     

Management of burn injury

Burns are a major cause of morbidity and mortality worldwide. Vulnerable people such as children, the frail and elderly, and the socially deprived are at particular risk. Most burns are caused by thermal injury to the skin, but electrical and chemical burns can be very severe. Fortunately, most burns are minor and superficial and can be managed by primary health care professionals. However, major and severe burns require in-hospital management from a team of surgeons and other specialists. Life-threatening conditions such as smoke inhalation airway damage and severe fluid loss should be addressed during the initial resuscitation. Prevention of further thermal damage by cooling is important along with prevention of secondary infection of burn injuries. A wide variety of dressings is available for the management of burns and expert nursing care is vital. Surgical intervention may be urgently required for fasciotomy or escharotomy in cases of compartment syndrome or circumferential burns, respectively. Debridement, skin grafting and reconstructive procedures will be required over the medium or long term for patients with severe or complex burns and should be planned with appropriate multidisciplinary expertize. The functional and psychological impact of major burn injury should not be underestimated.     

不同免疫调节剂对烫伤大鼠巨噬细胞功能的影响

目的探讨不同免疫调节剂对烫伤大鼠腹腔巨噬细胞(Mφ)功能的影响。方法应用 McAb APAAP 桥联酶标法、琼脂溶菌板法和 MTT 比色法等技术,分别测定了各组烫伤大鼠不同时相腹腔 Mφ各种功能的变化。结果①烫伤后大鼠腹腔 Mφ表面 Ia 抗原表达率降低,抗原提呈能力减弱,对念珠菌的噬菌率下降,溶菌酶活力减低,TNF 分泌增高,且伤后第10天比第5天更明显。以上结果与正常组比较,差异有非常显著意义(P<0.01)。②经不同免疫调节剂治疗后,各组烫伤大鼠腹腔 Mφ各种功能明显改善,各项指标与未用药的对照组比较,差异有非常显著意义(P<0.01)。结论烫伤后早期腹腔注射特异性免疫核糖核酸(iRNA)可明显改善伤鼠的免疫功能。     

Alterations of acute phase reaction and cytokine production in patients following severe burn injury

To determine the acute immunologic reaction, mediated by cytokines, interleukines (ILs) and growth factors and the susceptibility to infections and sepsis after severe burn injury a prospective, single unit, longitudinal study of acute phase reactants and mediators who performed. After approval by the ethics committee of our hospital, we investigated the plasma concentrations of IL-2, -6, -8, -10, and -13, the soluble IL-2 receptor (sIL-2R), and the acute phase proteins procalcitonin (PCT) and C-reactive protein (CRP) at admission and every 3 days in 24 patients over a time course of 28 days after thermal injury and categorized by percent burn: < or =30% (group 1; n=12) and >30% (group 2; n=12). Shortly after burn injury we found higher concentrations of IL-2, -6, -10 and PCT in those patients >30% TBSA. During the study period, we found significant higher levels of acute phase proteins, IL-6 and -8 in patients >30% TBSA. The incidence of SIRS and MODS was three times increased in patients >30% TBSA. Our results show different patterns of cytokines and acute phase proteins in patients with different burned surface areas over a long time and continuous monitoring of a more distinct inflammatory response in these patients.