大面积侵袭感染组织切除对烧伤创面脓毒症患者静息能量消耗的 …

OBJECTIVE: To evaluate the effect of extensive excision of invasive burn wound infection on hypermetabolic response in burn patients with sepsis. METHODS: Eight patients with major burn, complicated by invasive burn wound infection and sepsis, were consecutively admitted to our hospital from September 1997 to October 1998. Resting energy expenditures (REEs) were monitored by means of cardiorespiratory diagnostic system (Medical Graphics Corporation, USA) at patients' bedside. Plasma concentration of IL-6, IL-8, TNF-alpha, and LPS were assayed before and after surgical intervention and at the time when the patients' vital signs became stable. Correlation analysis between REEs and IL-6, IL-8, TNF-alpha, and LPS were made, respectively. RESULTS: A total of 8 treated patients survived. Values of REE before surgical intervention were significantly higher than those after surgical intervention (P < 0.01), and when patients' vital signs became stable the values were significantly lower than those after surgical intervention (P < 0.01). The plasma concentrations of IL-6, IL-8, TNF-alpha, and LPS after excision of invasive burn wound infection were significantly lower than those before surgical intervention (P < 0.05). The lowest levels of these inflammatory mediators were observed when the conditions of patients became stable, and the values were significantly lower than those before surgical intervention (P < 0.001). There was a significant positive correlation between REE level and respective values of plasma IL-6, IL-8, TNF-alpha, and LPS (P < 0.01). CONCLUSIONS: It seemed that the extensive excision of invasively infected burn wound in patients with major burn should be performed as early as possible to reduce an increased release of inflammatory mediators and to control the hypermetabolic response during sepsis.     

Clinical characteristics and treatment of burn wound sepsis in extensive burn patients: successful experience with eight cases

Eight burn wound sepsis patients, in which 6 cases were diagnosed as MODS and two as septic shock, were treated consecutively in our hospital from September 1997 to October 1998. The plasma concentration of IL-6, IL-8, TNFα and LPS were assayed before and after surgical intervention, as well as when the patients' vital signs became stable. The results showed: ①The patients' conditions abruptly deteriorated when the burn wound sepsis emerged;②The major cause related to burn wound sepsis was extensive burn injuries, with large areas of deep burn remaining open; ③Although wound swabs taken on admission revealed the presence of colonization by many pathogenic bacteria, Pseudomonas aeruginosa was one of the most frequent bacteria isolated from the subeschar tissue; ④The plasma concentrations of IL-6, IL-8, TNF and LPS before surgical intervention were significantly higher than that after surgical intervention (P＜0.05) ;⑤The lowest level of the inflammatory mediators was observed when the patients' conditions became stable, as compared with before surgical intervention (P＜0. 001).These findings suggest that the clinical characteristics of burn wound sepsis are abrupt deterioration of the general condition and prominent septic symptoms, often complicated by MODS. The main cause of burn wound sepsis is the presence of a large area of open deep burn wounds, which should be excised and covered early. LPS and pro-inflammatory mediators play an important role in the pathogenesis of burn wound sepsis. Although success in treating these patients is the result of appropriate application of multiple treatments, early, aggressive and thorough surgical excision of invasive burn infectious tissue and closure of wound play a crucial role in the successful treatment of patients complicated by burn wound sepsis. Other treatments are adjuvant but also important.     

大面积侵袭感染组织切除对烧伤创面脓毒症患者静息能量消耗的影响

目的观察大面积侵袭感染组织切除对烧伤创面脓毒症患者高代谢的影响。方法对连续救治的8例烧伤创面脓毒症患者,分别于大面积侵袭感染组织切除前,手术后和病情稳定时,对静息能量消耗(restingenergyexpenditure,REE)白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子α(TNFα)、内毒素(LPS)进行监测。结果侵袭感染组织切除后REE水平[(307.7±31.3)kJ·h     

不同免疫调节方式对烧伤脓毒症大鼠炎性反应的影响

Objective To investigate the effect of Thymosin and growth hormone(GH) on inflam-matory response in burn rats or burn rats with sepsis. Methods Sixty-four SD rats were randomly divided into normal control group (NC, without treatment), sepsis group (S, with injection of LPS), sepsis + Thy-mosin group (ST, with successive injection of Thymosin and LPS), sepsis + GH group [SGH, with succes-sive injection of recombinant human GH (rhGH) and LPS], burn group, burn + sepsis group (BS, with in-jection of LPS after burn), burn + sepsis + Thymosin group (BST, with successive injection of Thymosin and LPS after burn), burn + sepsis + GH (BSGH, with successive injection of rhGH and LPS after burn), with 8 rats in each group. Specimens of spleen tissues were harvested to determine HLA-DR in lymphocyte and e-valuate inflammatory cell infiltration (score). Specimens of peripheral blood were collected to determine Toll-like receptor 4 (TLR4) level in monocyte and serum level of TNF-α, IL-4, IL-6, IL-10. Results Compared with those in NC group, serum level of IL-10 in S group decreased obviously, while other indices increased obviously (P<0.01). The levels of HLA-DR and TLR4 and serum level of TNF-α were similar between SGH and ST groups (P>0.05). Compared with those in SGH group [(2.87±0.04) score, and IL-6 (0.0083±0.0018) μg/mg, IL-4 (0.0102±0.0021) μg/mg, IL-10 (0.0310±0.0027) μg/mg, re-spectively], degree of inflammatory cell infiltration (1.50±0.76) score and serum levels of IL-6, IL-4, IL-10 of rats in ST group decreased obviously (0.0064±0.0012, 0.0058±0.0024, 0.0230±0.0021 μg/mg, respectively, P<0.01). The levels of HLA-DR, TLR4 and inflammatory cell infiltration degree of spleen in B group were respectively higher than those in NC group and lower than those in BS group. Com-pared with those in NC group, serum levels of TNF-α, IL-6 in B group increased significantly, while IL-4, IL-10 showed an opposite tendency. There was no obvious difference between BST and BSGH groups in ser-um levels of HLA-DR and IL-6 (P>0.05). Compared with those in BST group, inflammatory cell infiltra-tion degree in spleen and the levels of TLR, TNF-α obviously decreased (P<0.01), while IL-4 and IL-10 levels increased in BSGH group (P<0.01). Conclusions Inhibitive effects between Thymosin and GH on extensive inflammatory reaction were similar with or without trauma, and GH has better effect as compared with Thymosin when with trauma.     

不同治疗阶段（1970～1998年）烧伤脓毒症的防治经验

目的　总结并提出伤后脓毒症临床研究及防治措施。　方法　将我科 1970年 1月至1998年 10月收治的 5 330例烧伤患者中 45 1例发生脓毒症者 ,分为 3个治疗阶段 ,结合临床监测技术的不断完善及一系列防治新方法的使用 ,分析烧伤后脓毒症发病率及死亡率降低的主要原因。　结果　第 3治疗阶段脓毒症总发生率 (6 .7% )和死亡率 (4 .5 % )以及烧伤面积大于 30 %TBSA伤员脓毒症的发生率和死亡率均显著低于第 1、2阶段 (P <0 .0 5～ 0 .0 1)。第 3阶段脓毒症并发MSOF死亡组IL - 6、IL - 8、LPS和TNFα ,在整个MSOF期间明显高于存活组 ,尤其是死亡前为甚 (P <0 .0 5～0 .0 1)。　结论　早期、适当、有效的复苏 ,及时防治伤后感染 ,对深度烧伤创面及早切削痂植皮 ,加强内脏器官的保护与支持是降低脓毒症发生率、提高治愈率的最重要措施。一旦发生烧伤创面脓毒症 ,尽早果断地切除坏死组织是成功救治的关键     

Modulation of inflammatory and catabolic responses in severely burned children by early burn wound excision in the first 24 hours

HYPOTHESIS: Early burn wound excision modulates the hypermetabolic response in severe pediatric burn injuries. DESIGN: Before-after trial. SETTING: A 30-bed burn referral center in a private, university-affiliated hospital. METHODS: We studied 35 severely burned children who were divided into 2 groups. One group (n = 20) was treated with early burn wound excision within 24 hours after the injury. The second group (n = 15) was treated conservatively with silver sulfadiazine in other burn facilities for 5 days, and burn wounds were surgically excised when patients were admitted to our burn center on day 6 after the injury. Data compiled included oxygen consumption and acute-phase protein, interleukin 1beta; interleukin 6, interleukin 10, tumor necrosis factor alpha, and anabolic hormone (growth hormone, insulinlike growth factor type 1) levels preoperatively and 24 hours and 5 days postoperatively. MAIN OUTCOME MEASURES: Acute-phase and hypermetabolic responses. RESULTS: Early burn wound excision abrogated the hypermetabolic response in pediatric burn patients. Patients who underwent conservative treatment had a significantly more severe inflammatory and hypermetabolic response at the same time interval and significantly lower levels of anabolic hormones. CONCLUSIONS: Early burn wound excision is a safe therapeutic approach that modulates the hypermetabolic response after burn injury. It was superior to the conservative treatment of silver sulfadiazine and delayed excision, and it should be considered when treating all severe full-thickness burns.     

早期切痂对重度烧伤患者静息能量消耗的影响

目的　应用代谢车动态观察烧伤患者早期切痂植皮对静息能量消耗 (REE)的影响。方法　根据首次切痂植皮时间 ,将 5 6例重度烧伤患者分为早期切痂植皮组 (A组 ,39例 ,伤后 5d内手术 )和非早期切痂植皮组 (B组 ,17例 ,伤后 5d后手术 )。首次切痂时将Ⅲ度及其周围深Ⅱ度创面全部切除 ,切痂后创面用异体皮加自体微粒皮覆盖。在治疗期间应用新型代谢车床旁动态监测患者REE变化 ,同时检测 9例A组患者和 7例B组患者的血浆白细胞介素 (IL)6、IL 8、肿瘤坏死因子α(TNF α)、内毒素 脂多糖 (LPS)等变化。　 结果　全部病例治愈 ,尽管两组患者REE均明显升高 ,但A组在切痂前、后及伤后 14d内REE明显低于B组 (P <0.0 5 )。A组患者血浆IL 6、IL 8、TNF α、LPS水平明显低于B组 (P <0.0 5 )。　 结论　大面积深度烧伤患者早期切痂植皮可在一定程度上降低机体的高代谢状态 ,其原因可能与及早封闭创面从而减少多种炎性介质的过度释放有关。     

烧伤后24小时内削痂对深Ⅱ度创面局部炎症反应的影响

目的探讨烧伤后24h内行削痂手术对深Ⅱ度创面局部炎症反应以及组织损害的影响。方法选择12例烧伤患者,伤后24h内对深Ⅱ度创面行削痂手术,将患者同一创面分为3个标本采集区,即手术前、手术后和未手术区。手术前标本取自削痂术前创面,手术后和未手术区创面标本于伤后5—7d获取。采用组织培养和比色法测定创面组织释放白细胞介素(IL)8、髓过氧化物酶(MPO)和丙二醛(MDA)的水平,通过HE和Masson染色对创面组织坏死程度进行形态学观察。结果削痂手术后,患者创面组织局部释放IL8、MPO、MDA的水平分别为(6.83±1.85)μg/L、(4.07±0.87)U/g、(8.94±5.66)μmol/g,与未手术创面比较明显降低(P<0.01)。形态学观察显示,手术前创面有凝固性坏死灶;未手术创面炎症反应明显,坏死组织范围扩大;削痂手术后创面局部的炎症反应得到改善,未见坏死组织范围扩大。结论伤后24h内行削痂手术,可以改善深Ⅱ度烧伤创面局部的炎症反应,防止创面进行性加深,有利于创面及早愈合。     

Excision of the burn wound in patients with large burns

The known major determinants of survival for patients with burn injury are age, burn size, inhalation injury, and infection. The clinical courses of 210 patients with burns of 30% of the body surface or greater treated from Jan 1, 1983, through Dec 31, 1985, were reviewed to determine whether excision of the burn wound could be identified as a factor in survival after massive burn injury. The predominant site of infection changed from the burn wound to the lung, with pneumonia being the most common bacterial infection in patients whose wounds were treated as described. Effective topical control of bacterial proliferation and excision of the burn wound have resulted in replacement of invasive bacterial wound infection by nonbacterial burn wound infection. The apparent advantages of excision, particularly in large burns, may reflect only patient selection, since only those patients who are considered to be physiologically stable and able to tolerate the physiologic stress of excision are considered for operation.     

早期肠内营养对烧伤后高代谢反应调节作用的研究

目的 观察早期肠内营养对烧伤高代谢反应的调节作用。方法 通过３０例烧伤患者用间接测热法监测静息能量消耗（ＲＥＥ）,监测血浆激素、脂类调节因子、细胞因子水平,并结合临床疗效和氮平衡变化来评价早期肠内营养的作用。结果 早期肠内营养（ＥＥＮ）组较延迟肠内营养（ＤＥＮ）组ＲＥＥ显著下降（Ｐ＜０.０５－０.０１）,升高的时间缩短（Ｐ＜０.０５）;ＥＥＮ组非蛋白呼吸商（ＮＰＲＱ）较ＤＥＮ组更接近生理状态,负氮平衡时间缩短。伤后第４天ＥＥＮ组血浆去甲肾上腺素（ＮＥ）、胰高糖素（ＧＬＵＣＡＧＯＮ）、前列腺素Ｅ２（ＰＧＥ２）、血栓素Ｂ２（ＴＸＢ２）、低于ＤＥＮ组（Ｐ＜０.０５）;伤后８－１２d肿瘤坏死因子α（ＴＮＦ－α）、白细胞介素－６（ＩＬ－６）显著低于ＤＥＮ组（Ｐ＜０.０５－０.０１）,胰岛素水平呈相反变化。结论 严重烧伤后尽早开始肠内营养对高代谢反应有重要的调节作用。     

影响烧伤病人疗程的相关因素分析

目的探讨如何减少烧伤病人治愈时间,缩短住院日,降低病人的治疗费用。方法通过应用主成分分析的统计方法对我科７９１例病人进行回顾,试图找出与病人治愈时间密切相关的因素。结果烧伤面积的大小及深度、吸入性损伤、脓毒症、休克、内脏并发症、复合伤或合并伤和切痂面积、切痂时间、休克期切痂与否、抗生素应用时间等均是影响烧伤病人治愈时间和住院日的主要因素,进一步分析表明,休克期切痂病人内脏并发症及脓毒症的发生率低,治愈时间短,尤其是超过３０％烧伤面积者,休克期切痂组病人治愈时间明显少于非休克期切痂组病人。结论积极防治休克、脓毒症及内脏并发症的发生发展,合理应用抗生素等,而更重要的是大面积深度烧伤创面早期切削痂植皮等均是缩短病人治愈时间的有效手段。     

环氧合酶2对脓毒症大鼠免疫功能的影响

目的 研究环氧合酶2(cyclooxygenase-2,COX-2)在脓毒症大鼠肝组织炎症反应中的表达及其对免疫功能的影响.方法 选择质量相近的Wistar大鼠54只,分为假手术组(6只)、脓毒症组(24只)、NS-398干预组(24只).通过建立盲肠结扎穿孔脓毒症动物模型,用RT-PCR方法检测肝组织COX-2mRNA的表达,ELISA法检测血清IL-6、IL-10、TNF-α水平,流式细胞仪检测T淋巴细胞亚群CD4+、CD8+细胞的变化,同时观察肝脏病理变化.结果 (1)脓毒症组病理损伤重,NS-398干预组损伤减轻.(2)COX-2mRNA:假手术组呈低表达,脓毒症组、NS-398干预组升高,但NS-398干预组较脓毒症组明显偏低.(3)IL-6:脓毒症组较假手术组和NS-398干预组明显增高(F=125.582,134.712,54.760,121.441,P＜0.05).(4)IL-10:NS-398干预组高于假手术组和脓毒症组(F=39.064,34.382,51.115,8.174,P＜0.05).(5)TNF-α:脓毒症组、NS-398干预组增高,但两组比较差异无统计学意义(x2=5.600,6.162,7.136,7.200,P＞0.05).(6)CD4+/CD8+比值:NS-398干预组高于脓毒症组(F=17.448,15.055,30.068,64.210,P＜0.05).结论 COX-2在脓毒症中发挥重要作用,其作用途径可能是通过改变促炎、抗炎细胞因子之间以及CD4+、CD8+细胞之间的平衡,使机体免疫状态紊乱而实现的.     

影响烧伤病人疗程的相关因素分析

目的 探讨如何减少烧伤病人治愈时间，缩短住院日，降低病人的治疗费用。方法 通过应用主成分分析的统计方法对我科７９１例病人进行回顾，度图找与病人治愈时间密切相关的因素。结果 烧面积的大小及深度、吸入性听凭、脓毒症、休克、内脏并发症、复合伤或合并伤和切痂面积、切痂时间、休克期切痂与否、抗生素应有理影响烧伤病人治愈时间和住院日的主要因素，进一步分析表明，休克期切痂病人内脏并发症及脓症的发生率低，治愈时间     

对烧伤感染的认识

Burn infection occurs when pathogenic bacte-ria colonized on the burn wound surfate,and they then invaded the viable tissue causing sepsis or sepsis with blood stream inva-sion.This infection pattern is particular to burn injury.Both in a model of pseudomonas burn wound sepsis and a clinical study of early eschar excision for bacteria quantification indicate that the bacteria not only are located on the burn wound surface but also invaded the deeper tissues.Finally,the bacteria penetrate into the neighboring viable tissue and even blood ves-sels.Therefore,we can say that burn infection is from local wound infection to invasive infection.and finally sepsis is devel-oped,and it is termed as burn wound sepsis.The cutoff count of subeschar tissue bacteria is 105/g.However,the burn wound sepsis may not occur when the number of subeschar tissue bacteria reaches 105/g.The criteria for the diagnosis of burn wound sepsis are mainly listed as below:(1)The number of bacteria in the subes-char reaches≥105/g.(2)Bacteria can be detected in the biopsy specimen.(3)Sepsis associated symptoms and signs.However,the sepsis associated symptoms and signs must be obvious in patients to make the clinical diagnosis of burn wound sepsis.If the sepsis associated symptoms and signs do not ap-pear.we should not make the diagnosis of burn wound sepsis e-ven with the number of bacteria in the subeschar tissue reaching 105/g or bacteria can be found in the biopsy specimen.Sepsis has been defined as the body's response to bacteria and their products.The occurrence of sepsis depends primarily on immune function and stress response intensity.and it is closely related to wound infection degree such as bacteria density and invasion depth in the burn wound,or plasma endotoxin level to certain extent.     

抗内毒素Fab'对严重烧伤早期炎症细胞因子的影响

目的 观察抗内毒素Fab'对严重烧伤早期肠源性内毒素血症小鼠肠道损伤的保护作用.方法 采用严重烧伤早期肠源性内毒素血症小鼠模型,分为烧伤组、治疗组及对照组,分别于6、12、24、48 h四个时相点测定血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-lβ、IL-10的浓度.结果 与正常对照组比较,烧伤后血清TNF-α、IL-1β、IL-10水平增高,差异有统计学意义(P<0.01);治疗组血清TNF-α、IL-1β、IL-10水平较烧伤组显著降低(P<0.01).病理检查结果提示治疗组较烧伤组肠黏膜损伤明显减轻.结论 抗内毒素Fab'能抑制内毒素所诱导的TNF-α、IL-1β产生,同时调节血清中的IL-10水平,减轻内毒素对机体的损害,从而起到对严重烧伤后肠源性脓毒症的防治作用.     

Effects of early excision and aggressive enteral feeding on hypermetabolism,catabolism, and sepsis after severe burn

BACKGROUND: Severe burn induces a systemic hypermetabolic response, which includes increased energy expenditure, protein catabolism, and diminished immunity. We hypothesized that early burn excision and aggressive enteral feeding diminish hypermetabolism. METHODS: Forty-six burned children were enrolled into a cohort analytic study. Cohorts were segregated according to time from burn to transfer to our institution for excision, grafting, and nutritional support. No subject had undergone wound excision or continuous nutritional support before transfer. Resting energy expenditure, skeletal muscle protein kinetics, the degree of bacterial colonization from quantitative cultures, and the incidence of burn sepsis were measured as outcome variables. RESULTS: Early, aggressive treatment did not decrease energy expenditure; however, it did markedly attenuate muscle protein catabolism when compared with delay in aggressive treatment. Wound colonization and sepsis were diminished in the early treatment group as well. CONCLUSION: Early excision and concurrent aggressive feeding attenuate muscle catabolism and improve infectious outcomes after burn.     

Effects of delayed wound excision and grafting in severely burned children

HYPOTHESIS: Advances in burn treatment including early excision of the wound have increased survival in patients treated at specialized burn centers. We hypothesized that the patients with delayed wound excision and grafting would experience deleterious outcomes. METHODS: From 1995 to 1999, 157 children with acute burns covering 40% or more of total body surface area and having more than 10% of full-thickness burns were admitted to our institution within 2 weeks of injury. Among them, 86, 42, and 29 patients underwent first operation on days 0 to 2, days 3 to 6, and days 7 to 14 after burn, respectively. Outcomes observed were mortality, number of operative procedures, length of hospitalization, blood transfused, incidence of wound bacterial and fungal contamination, invasive wound infection, and sepsis. RESULTS: Demographic data for the groups showed no differences in sex or total body surface area burned. Mortality and number of operative procedures and blood transfusions were not different between groups. Hospitalizations were longer in the delayed groups, which was associated with a higher incidence of significant wound contamination (P =.008). Invasive wound infection also increased significantly with delay of excision (P<.001). An increased incidence of sepsis was seen in patients with delayed wound excision and grafting (P =.04). CONCLUSIONS: Delays in excision were associated with longer hospitalization and delayed wound closure, as well as increased rates of invasive wound infection and sepsis. Our data indicate that early excision within 48 hours is optimal for pediatric patients with massive burns.     

影响烧伤病人疗程的相关因素分析

目的探讨如何减少烧伤病人治愈时间,缩短住院日,降低病人的治疗费用。方法通过应用主成分分析的统计方法对我科791例病人进行回顾,试图找出与病人治愈时间密切相关的因素。结果烧伤面积的大小及深度、吸入性损伤、脓毒症、休克、内脏并发症、复合伤或合并伤和切痂面积、切痂时间、休克期切痂与否、抗生素应用时间等均是影响烧伤病人治愈时间和住院日的主要因素,进一步分析表明,休克期切痂病人内脏并发症及脓毒症的发生率低,治愈时间短,尤其是超过30%烧伤面积者,休克期切痂组病人治愈时间明显少于非休克期切痂组病人。结论积极防治休克、脓毒症及内脏并发症的发生发展,合理应用抗生素等,而更重要的是大面积深度烧伤创面早期切削痂植皮等均是缩短病人治愈时间的有效手段。     

The effects of early excision and grafting on myocardial inflammation and function after burn injury

BACKGROUND: Sepsis is a frequent complication of burn injury despite absence of confirmed infection. Numerous investigators have proposed that the burn wound itself is a primary stimulus for postburn inflammation, and that early excision of the burn wound attenuates the hypermetabolic and inflammatory responses to burn injury. However, others have suggested that aggressive fluid resuscitation and correction of postburn fluid and electrolyte deficits should be the primary focus of intervention in the first 24 hours postburn. This present study determined whether excision and grafting of the burned wound within 30 minutes after injury abrogated myocardial inflammation and contractile defects that occur after burn injury. METHODS: In group 1, Sprague Dawley rats were given a third-degree burn over 20% total body surface area (TBSA), whereas rats in group 2 had burns over 30% TBSA and no wound excision. Rats in groups 3 and 4 had burn over 20% and 30% TBSA, respectively, followed by-wound excision and grafting (WE/G) within 30 minutes after completing burn injury. Group 5 included sham burn with no excision, whereas rats in groups 6 and 7 included shams that had either 20% or 30% normal skin excised and grafted to provide appropriate surgical controls. All rats received lactated Ringer's (4 mL/kg/% burn or percent wound excision). Twenty-four hours postburn, hearts were perfused (Langendorff) to assess ventricular function; myocytes were isolated to examine cytokine secretion and Ca2+/Na+ homeostasis. RESULTS: Burn in the absence of wound excision produced myocardial inflammation and contractile defects as indicted by a lower left ventricular pressure and lower rate of left ventricular pressure rise (+dP/dt) and fall (-dP/dt) response to maximal increases in preload or perfusate Ca2+ compared with responses measured in sham hearts. WE/G within 30 minutes after burn injury reduced myocyte secretion of proinflammatory cytokines and improved left ventricular pressure and +/-dP/dt responses to inotropic challenge. CONCLUSION: Cutaneous burn injury and the loss of the skin barrier function contribute, in part, to the myocardial inflammation which, in turn, contributes to myocardial contractile dysfunction that is characteristic of major burn injury.     

肠三叶因子和黏蛋白对烧伤血清所致肠上皮细胞免疫功能变化的影响

目的 观察肠三叶因子(ITF)和黏蛋白对烧伤血清所致肠上皮细胞免疫功能变化的影响.方法 (1)体外培养大鼠小肠上皮细胞株IEC-6,根据培养液添加物质不同,按照随机数字表法将细胞分为5组:①正常对照组,培养液中含10％(指体积分数,下同)小牛血清;②烧伤对照组,培养液含10％烧伤血清;③ITF+烧伤血清组,培养液含10％烧伤血清及终浓度25μg/mL ITF;④黏蛋白+烧伤血清组,培养液含10％烧伤血清及终浓度250 μg/mL黏蛋白;⑤ITF+黏蛋白+烧伤血清组,培养液含10％烧伤血清及终浓度25 μg/mL ITF、250 μg/mL黏蛋白.向各组细胞加入上述培养液的同时,加入大肠杆菌菌液(1×108 CFU/mL,200 μL).继续培养15 min、30 min、1h、2h、3h后,行瑞氏-吉姆萨染色,于显微镜下观察并统计黏附在细胞上的细菌数;采用锥虫蓝染色法观察并统计细胞存活率.每组每时相点样本数均为20.(2)将IEC-6细胞按照随机数字表法分为4组:烧伤对照组、ITF+烧伤血清组、黏蛋白+烧伤血清组、ITF+黏蛋白+烧伤血清组,分别同前加入相应的培养液(不加菌液)培养3、6、12、24、48 h.采用放射免疫分析法测定各时相点培养上清液中TNF-α、IL-6和IL-8 含量,每组每时相点样本数均为6.对实验数据行t检验.结果 (1)烧伤对照组细胞各时相点细菌黏附数量较正常对照组明显增多(t值为2.947 ～8.149,P值均小于0.01).与烧伤对照组比较,其余3个烧伤血清组在加菌后多数时相点细菌黏附的数量明显偏少(t值为-4.733～-2.180,P＜0 05或P ＜0.01).烧伤对照组细胞各时相点存活率与正常对照组比较均明显降低(t值为-4.126～-2 363,P值均小于0 05).ITF+烧伤血清组、黏蛋白+烧伤血清组细胞存活率在部分时相点明显高于烧伤对照组(t值为2.120～3.423,P＜0.05或P＜0.01).ITF+黏蛋白+烧伤血清组细胞存活率加菌后15 min为(96.7±2 4)％,明显高于ITF+烧伤血清组[(94 5±3 1)％,t=2.507,P＜0.05];在3h时为(84.0±6 7)％,明显高于黏蛋白+烧伤血清组[(77 1±8 2)％,t=2.934,P＜0.01].(2)ITF+黏蛋白+烧伤血清组培养6、12、24、48 h,TNF-α含量均低于其余3组(t值为-6.914 ～ -2.889,P＜0.05或P＜0.0i).ITF+黏蛋白+烧伤血清组IL-6含量在部分时相点明显低于其余3组(t值为-7.657～-2.580,P＜0.05或P＜0.01).ITF+黏蛋白+烧伤血清组在培养6、12、24、48 h IL-8含量明显低于烧伤对照组和黏蛋白+烧伤血清组(t值为-8.802 ～ -3.640,P值均小于0.01);在培养12、24 h明显低于ITF+烧伤血清组(t值分别为-2.786、-2.740,P值均小于0.05).结论 ITF能维护肠上皮细胞功能,抵御细菌黏附,降低细胞死亡率,同时能维护细胞免疫稳态,减少炎症介质释放;ITF与黏蛋白联用效果更明显.