The effect of halothane and isoflurane on plasma cytokine levels

The aim of this study was to investigate the effect of halothane vs. isoflurane on cytokine production during minor elective surgery. Forty adult patients, ASA I-II were randomly allocated to receive halothane or isoflurane. Venous samples for interleukin (IL)-1beta, IL-2, IL-6, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were taken before anaesthesia, before incision, at the end of anaesthesia and 24 h postoperatively. In both groups, IL-6 and TNF-alpha levels remained low throughout the study period. Before incision, in both groups IL-1beta and IFN-gamma showed a decrease (p<0.01 for IL-1beta in isoflurane group and p<0.05 for the others) compared with pre-induction. By the end of anaesthesia and surgery, IL-1beta had increased significantly (p<0.05) and IFN-gamma had decreased significantly (p<0.05) in both groups compared with pre-incisional levels. By 24 h postoperatively in both groups, IL-1beta had decreased significantly (p<0.05), whereas IFN-gamma had increased significantly (p<0.05) compared with the end of anaesthesia and surgery level. Pre-incisionally, IL-2 increased in the halothane group (p<0.01), whereas it decreased significantly in the isoflurane group (p<0.001) compared with the pre-induction level. By the end of anaesthesia and surgery and by 24 h postoperatively, IL-2 had decreased significantly in the halothane group (p<0.001), whereas it increased significantly in the isoflurane group (p<0.001) compared with pre-incision and end of anaesthesia and surgery levels, respectively.     

A comparison of the effect of total intravenous anaesthesia with propofol and remifentanil and inhalational anaesthesia with isoflurane on the release of pro- and anti-inflammatory cytokines in patients undergoing open cholecystectomy

The aim of the study was to investigate the effects of two anaesthetic techniques (total intravenous technique vs. inhalational technique) on changes in pro- and anti-inflammatory cytokine levels during open cholecystectomy. Forty ASA PS I-II patients undergoing open cholecystectomy were randomly assigned to two groups. Group R received total intravenous anaesthesia with propofol and remifentanil and group F received balanced inhalational anaesthesia with isoflurane. The plasma levels of tumour necrosis factor-alpha (TNF-alpha), interleukin IL-6 and interleukin IL-10 were measured during and after surgery. The pro-inflammatory cytokine levels (TNF-alpha and IL-6) and the anti-inflammatory cytokine (IL-10) showed a significant increase in their concentrations compared with pre-induction levels in both groups (P < 0.05). By the end of anaesthesia and surgery, TNF-alpha and IL-6 were significantly lower in group R than in group F (P < 0.05). At the end of anaesthesia and 12 hours postoperatively, IL-10 levels in group R were higher than in group F (P < 0.05). These findings suggest that total intravenous anaesthesia using propofol and remifentanil suppresses the inflammatory response caused by surgery to a greater extent than a balanced inhalation technique using isoflurane.     

The immunomodulatory effects of prolonged intravenous infusion of propofol versus midazolam in critically ill surgical patients

Both propofol and midazolam are known to inhibit immune function. The aim of this study was to investigate cytokine production in critically ill surgical patients as early markers of immune response to prolonged infusion of propofol and midazolam. The study enrolled 40 elective patients who were to receive long-term sedation for more than 2 days. Patients were randomly allocated to one of two equally sized groups. Central venous blood samples for measurement of interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were drawn prior to the start and after 48 h of infusion. After 48 h, propofol caused significant increases in IL-1beta (24%), IL-6 (23%) and TNF-alpha (4.8 times) levels, while midazolam caused significant decreases in IL-1beta (21%), IL-6 (21%) and TNF-alpha (19%). Both agents caused significant decreases in IL-8 levels (propofol: 30%, midazolam: 48%, p < 0.05). Propofol caused significant decreases in IL-2 levels (68%, p < 0.001) but increases in IFN-gamma (30%, p < 0.05), whereas there was no significant change with midazolam compared with the pre-infusion level. In conclusion, during 48 h of continuous infusion, propofol stimulated, while midazolam suppressed, the production of the pro-inflammatory cytokines IL-1beta, IL-6 and TNF-alpha, and both caused suppression of IL-8 production. Propofol inhibited IL-2 production and stimulated IFN-gamma production, whereas midazolam failed to do so. Therefore, sedative agents may have clinical implications in high-risk and immunocompromised patients.     

Aortic aneurysm repair is associated with a lower inflammatory response compared with surgery for inflammatory bowel disease

BACKGROUND: Since the plasma cytokine profile reflects the body's inflammatory response to injury, this study was designed to prospectively observe the plasma cytokine levels in response to the degree of different sorts of abdominal surgical trauma. METHODS: Plasma levels of TNF-alpha, type I TNF receptor (p55), type II TNF receptor (p75), IL-6, IL-8, IL-10, phospholipase A(2) (PLA(2)), and haptoglobin were measured peri-operatively in patients undergoing bowel resection for inflammatory bowel disease or diverticulitis (IBD) (n = 9), elective repair of abdominal aortic aneurysm (AAA) (n = 9), or laparoscopic cholecystectomy (lap chole) (n = 9). RESULTS: The IBD patients showed a significant (p < 0.05) post-operative elevation in plasma IL-6, p55, p75, and PLA(2) levels, but no significant change in TNF-alpha, IL-8, IL-10 or haptoglobin levels. The AAA patients had a significant post-operative rise in IL-10 levels and a significant decrease in plasma haptoglobin levels, but no significant change of TNF-alpha, IL-6, IL-8, p55, p75, or PLA(2) concentrations. The lap chole patients demonstrated no significant change in any of these parameters. CONCLUSION: These data show that IL-6, IL-10, p55, and p75 are markers to measure the degree of inflammatory stress associated with abdominal operative procedures and demonstrate the relative lack of a cytokine response to laparoscopic cholecystectomy.     

Comparison of immunologic and physiologic effects of CO2 pneumoperitoneum at room and body temperatures

BACKGROUND: Prolonged and complex laparoscopic procedures expose patients to large volumes of cool insufflation gas. The aim of this study was to compare the effects of a conventional room temperature carbon dioxide (CO2) pneumoperitoneum with those of a body temperature pneumoperitoneum. METHODS: Patients were randomized to undergo laparoscopic cholecystectomy with a CO2 pneumoperitoneum warmed to either body temperature (n = 15) or room temperature (n = 15). The physiologic and immunologic effects of warming the gas were examined by measuring peroperative core and intraperitoneal temperatures, peritoneal fluid cytokine concentrations, and postoperative pain. RESULTS: The mean duration of surgery was 32 min in both groups. Core temperature was reduced in the room temperature group (mean, 0.42 degrees C; p < 0.05). No reduction in temperature occurred when the gas was warmed. Greater levels of cytokines were detected in peritoneal fluid from the room temperature insufflation group tumor necrosis factor alpha (TNF-alpha): mean, 10.9 pg/ml vs. 0.42, p < 0.05; interleukin 1 beta (IL-1beta): mean, 44.8 pg/ml vs. 15.5, p < 0.05; and IL-6: mean, 60.4 ng/ml vs. 47.2. There was no difference in postoperative pain scores or analgesia consumption between the two groups. CONCLUSIONS: The authors conclude that intraoperative cooling can be prevented by warming the insufflation gas, even in short laparoscopic procedures. In addition, warming the insufflation gas leads to a reduced postoperative intraperitoneal cytokine response.     

Downregulation of T helper type 1 immune response and altered pro-inflammatory and anti-inflammatory T cell cytokine balance following conventional but not laparoscopic surgery

BACKGROUND: The clinical advantages of laparoscopic procedures result from a minimized surgical trauma. The present study was performed to investigate immunosupression following laparoscopic operations as compared with open surgery. Our analysis focused on the T cell secretion of cytokines that regulate the critical balance of either T helper type-1 (Th1)- and Th2-mediated immune responses on pro- and antiinflammatory activities. METHODS: In a prospective study, immunological data of 26 patients submitted to laparoscopic cholecystectomy (LCE) and 17 patients undergoing conventional cholecystectomy (CCE) for symptomatic cholecystolithiasis were compared. Patients with acute cholecystitis and patients developing postoperative complications or receiving immunosuppressive medication were excluded. Production of interferon (IFN)-gamma, interleukin (IL)-2, IL-4, tumor necrosis factor (TNF)-alpha, and IL-10 by isolated T cells stimulated by cross-linking of CD3 and CD28 was evaluated preoperatively as well as on postoperative days 1 and 6 or 7. Cytokines were measured by immunoenzymometric assay. RESULTS: IFN-gamma, TNF-alpha, and IL-2 production by T cells decreased significantly by 48.3%, 36.6%, and 36.8%, respectively, on postoperative day 1 after CCE, but not after LCE. These results indicate severe suppression of Th1-type and proinflammatory cytokines after the open operation. In contrast, IL-4 and IL-10 did not show significant changes in either group suggesting that Th2 cell response and anti-inflammatory activity remained normal. CONCLUSIONS: The present study shows that open, but not laparoscopic cholecystectomy is associated with a marked suppression of T lymphocytes functions as indicated by deregulation of both the Th1/Th2 and the pro-/anti-inflammatory cytokine balance. The results therefore suggest that downregulation of Th1 cell-mediated immune response and pro-inflammatory activity of T cells is a hallmark of open, but not laparoscopic surgery.     

Immunparalyse von T-Lymphocyten und Monocyten in der postoperativen abdominellen Sepsis

In vitro functions of stimulated peripheral T cells and monocytes were investigate in patients experiencing sepsis following major visceral surgery. Cell culture supernatants were analyzed by ELISA for IL-2, IFN-gamma, IL-4, IL-10, TNF-alpha, IL-1 beta, and IL-12p40. In addition, monocyte HLA class II expression was determined by flow cytometry. T cell secretion of IL-2, TNF-alpha, and in part IFN-gamma (but not IL-4) was significantly diminished in non-survivors throughout the entire course of sepsis, compared to controls and sepsis survivors. Production of IL-1 beta and IL-12 p40 by monocytes was strongly reduced in both survivors and non-survivors at the onset of sepsis. Persistence of depressed monocyte cytokine secretion correlated with lethality. Thus, overall suppression of cytokine production by T cells and monocytes was already observed at the beginning of postoperative sepsis. HLA class II expression by monocytes exhibited a strong and sustained down-regulation with no significant differences between sepsis survivors and non-survivors. In summary, suppression of both T cell and monocyte functions develops early during postoperative sepsis. Recovery of immune functions and severity of immune defects are associated with outcome.     

不同麻醉方式对炎性细胞因子的影响

目的比较雷米芬太尼-丙泊酚全凭静脉麻醉与异氟醚静吸复合全身麻醉用于胆囊切除术对促炎性和抗炎性细胞因子的影响。方法慢性胆囊炎或胆囊结石患者28例,ASAⅠ或Ⅱ级,随机均分为全凭静脉麻醉组(T组)和静吸复合麻醉组(I组),测定两组患者麻醉前、术后4、12h血浆细胞因子白细胞介素-6(IL-6)和白细胞介素-10(IL-10)水平。结果两组血浆IL-6、IL-10水平在术后4h均显著升高(P<0.05)。术后12hT组血浆IL-6水平显著低于I组(P<0.05)。结论雷米芬太尼-丙泊酚全凭静脉麻醉可增强抗炎细胞因子反应,有利于减轻手术应激的炎症反应。     

Systemic and cell-mediated immune response after laparoscopic and open cholecystectomy in patients with chronic liver disease. A randomized, prospective study

BACKGROUND: As impaired immune function observed in cirrhotic patients is known to increase the risk of postoperative complications, the immunological response to surgery was investigated. METHODS: Twenty-eight patients with postnecrotic liver cirrhosis or chronic hepatitis C and symptomatic gallstone disease were randomly allocated to laparoscopic (LC) or open cholecystectomy (OC). Changes in concentrations of cytokines (TNF-alpha, IL-1beta, IL-6, IL-8 and IL-10) were followed and the effect of surgical trauma on the distribution of lymphocyte subpopulations (CD3, CD4, CD8, CD16 and CD19) and NK cell cytotoxicity were measured. RESULTS: After OC a decrease in circulating CD3 (p < 0.05) and CD4 (p < 0.05) and an increase in CD19 (p < 0.05) cells were detected in contrast to LC after which only CD16 cells decreased (p = 0.05). The number of CD3 cells was higher after LC than after OC (p < 0.01), whereas the number of CD19 cells was higher after OC than after LC (p < 0.01). NK cell cytotoxicity was reduced after LC (p < 0.05). In cirrhotic patients circulating cytokines were unaffected by OC, whereas TNF-alpha (p < 0.05) and IL-1beta (p < 0.05) were reduced after LC. In chronic hepatitis IL-1beta decreased after OC (p = 0.05) and IL-10 was significantly higher after LC than following OC (p < 0.05). CONCLUSION: The immune response is less pronounced after a laparoscopic procedure compared to a conventional approach in patients with chronic liver disease.     

Hormone-cytokine response

Background: Changes in blood hormone and cytokine were investigated in patients who underwent laparoscopic cholecystectomy via insufflation (CO₂ group) vs those who had abdominal wall-lifting (Air group). Methods: Seventeen female patients with cholecystolithiasis were randomly divided into two groups. Peripheral blood samples were obtained during perioperative period, and plasma hormone levels (ACTH, cortisol) and serum cytokine levels (TNFα, IL-1β, IL-6, IL-10) were measured. Results: The number of circulating lymphocytes significantly decreased at 1 h after surgery in both groups, but the decrease in the CO₂ group was significantly smaller than that in the Air group. There was no significant difference in hormone elevation between groups. Serum concentrations of IL-6 and IL-10 in the Air group were significantly higher than in the CO₂ group. Conclusions: CO₂ insufflation may reduce cytokine production in laparoscopic cholecystectomy. Received: 10 November 1996/Accepted: 19 February 1997     

Paediatric laparoscopic surgery: anaesthetic management

Summary
Total intravenous anaesthesia (TIVA) has been performed in 54 paediatric patients undergoing laparoscopic surgery. A loading dose of propofol 2.5 mg·kg⁻¹ and fentanyl 2 μg·kg⁻¹ were used for induction while a continuous infusion of propofol 9 mg·kg⁻¹·h⁻¹ and atracurium 0.5 mg·kg⁻¹·h⁻¹ in O₂/Air were used for maintenance. Intraoperative cardiorespiratory stability, prompt recovery, painless postoperative period show that TIVA is a valid and safe technique for laparoscopic surgery in children.     

Changes in blood coagulation, fibrinolysis and cytokine profile during laparoscopic and open cholecystectomy

The aim of this prospective, non-randomised study was to investigate haemostatic system alterations in patients undergoing open (OC) and laparoscopic cholecystectomy (VLC). In addition, we also measure the plasma cytokine profile to explore any relationship between changes in plasma cytokine levels and the postoperative coagulation profile. From July 2005 to March 2007, 71 patients were non-randomly assigned to open (group 1) or laparoscopic cholecystectomy (group 2). Prothrombin fragment 1.2 (F1.2), thrombin-antithrombin (TAT), fibrinogen, soluble fibrin, antithrombin III (AT), protein C, plasminogen and D-dimer levels were measured at baseline and at 1, 24, 48 and 72 hours postoperatively. Serial serum levels of IL-1 beta and IL-6 were measured by colorimetric ELISA. Plasma levels of F1.2, TAT, fibrinogen, soluble fibrin and D-dimer increased significantly in group 1. Plasma levels of AT, protein C, plasminogen decreased in both groups. In the OC group, the serum IL-1 beta and IL-6 levels began to increase significantly as early as 1 hour after the start of the operation, peaking at hour 6. The surge in circulating cytokine levels, commonly found in the postoperative period, is shown to be capable of inducing a hypercoagulability state and there is a positive correlation between IL-6 levels and hypercoagulability. In our study only mild hypercoagulability was observed in patients undergoing laparoscopic cholecystectomy. In conclusion, the correlation between cytokine levels and coagulation activation may be related to the type of surgery performed. Our present knowledge of the effect of laparoscopy upon coagulation and fibrinolysis is incomplete and based on only a few studies; for this reason further studies are required to investigate these aspects.     

腹腔镜结直肠癌手术对机体影响的探讨

目的探讨腹腔镜下结直肠癌手术的低侵袭性。方法将符合纳入研究对象标准的40例结直肠癌患者随机分成腹腔镜组(20例)和开腹组(20例),比较两组患者围手术期(术前、术后当天、术后第1、3、5d)的外周血白介素(IL)-6、IL-8、肿瘤坏死因子(TNF)-α、C反应蛋白(CRP)、可溶性细胞间黏附分子(sICAM-1)、白细胞CD11b的变化。结果开腹组术后细胞因子(TNF-α、IL-6、IL-8)明显高于腹腔镜组(P<0.05)。开腹组术后6h、第1天时,sICAM-1的动态变化较腹腔镜组显著升高,开腹组外周血白细胞CD11b在术后6h降至最低(161.98±48.42),较腹腔镜组(189.51±46.45)明显低(P<0.05)。结论结直肠癌的腹腔镜手术比传统开腹手术对机体影响小,具有明显的低侵袭性。     

Intracellular monocyte cytokine production and CD 14 expression are up-regulated in severe vs mild chronic heart failure.

BACKGROUND: The role of circulating monocytes in the process of low-grade inflammation, characteristic of chronic heart failure (CHF), has recently been questioned. Lipopolysaccharide (LPS) desensitization has been proposed to mediate reduced monocyte cytokine elaboration in patients with severe CHF. METHODS: Intracellular monocyte production of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor (TNF)-alpha, and monocyte CD 14 expression were measured flow-cytometrically without and after 8-hour LPS stimulation in 46 patients with CHF and in a healthy control group. RESULTS: Basal cytokine concentrations were similar for the control and the mild CHF groups (New York Heart Association [NYHA] Class I or II). After LPS stimulation, IL-6 (p=0.002) and TNF-alpha levels (p=0.001) were lower in the latter group, whereas IL-1 beta production was comparable. For the moderate-severe CHF patients, unstimulated IL-1 beta (p=0.04) was higher, whereas IL-6 (p=0.2) and TNF-alpha (p=0.1) levels were not different from the controls. Measurement of LPS-stimulated cytokine production showed no differences between the control group and patients with moderate-severe CHF (all p= 0.5). Upon comparing mild vs moderate-severe CHF patients, higher levels of unstimulated cytokine production (IL-1 beta, p=0.002; IL-6, p=0.01; TNF-alpha, p=0.003), stimulated IL-1 beta (p=0.002) and IL-6 (p=0.008) were found in the latter patients. CD 14 expression in the moderate-severe CHF group was higher than in the mild-CHF group (p = 0.03) and was strongly related to stimulated IL-1 beta (r=0.62, p<0.0001), IL-6 (r=0.56, p=0.0002) and TNF-alpha (r=0.41, p=0.006) production. CONCLUSIONS: CD 14 expression and monocyte cytokine production, both unstimulated and after LPS stimulation, are increased in moderate-severe CHF when compared with mild CHF. These data suggest that circulating monocytes, possibly via increased CD 14 expression, may play a significant role in the immunologic dysbalance observed in advanced CHF.     

Changes in the blood coagulation,fibrinolysis, and cytokine profile during laparoscopic and open cholecystectomy

Background: It has long been known that a hypercoagulability state develops after surgery. A surge in circulating cytokine levels is also commonly found in the postoperative period. These cytokines have all been shown to be capable of inducing a hypercoagulability state. Recently laparoscopic cholecystectomy (LC) has been introduced, and its advantages over the open procedure seem related to the reduced surgical trauma. LC is associated with a diminished acute-phase response compared with the open procedure. Our present knowledge on the influence of laparoscopic upon coagulation and fibrinolysis is incomplete and based on a few studies. Methods: The aim of this prospective, nonrandomized study was to investigate hemostatic system alterations in patients who undergo open and laparoscopic cholecystectomy. In addition we also measured the plasma cytokine profile to explore any relationship between changes in plasma cytokine levels and postoperative coagulation profile. Between September 1999 and April 2002, 71 patients were nonrandomly assigned to open (group 1) or laparoscopic cholecystectomy (group 2). All patients from group 1 were operated by a surgical team different from ours, who prefers the OC procedure. The patients with acute cholecystitis were excluded. Prothrombin fragment 1.2 (F1.2), thrombin-antithrombin (TAT), fibrinogen, soluble fibrin, antithrombin III (AT), protein C, plasminogen, and D-dimer levels were measured at baseline and at 1, 24, 48, and 72 h postoperatively. Serial serum levels of IL-1 and IL-6 were measured by colorimetric enzyme-linked immunosorbent assay (ELISA). Results: Plasma levels of F1.2, TAT, fibrinogen, soluble fibrin, and D-dimer increased significantly in group 1. Plasma levels of AT, protein C, and plasminogen decreased in both groups. In the OC group, the serum IL-3 and IL-6 levels began to significantly increased as early as 1 h from the beginning of the operation, revealing a peak at the sixth hour. When IL-6 and IL-1 levels were markedly elevated also, F1.2, fibrinogen, and soluble fibrin levels were increased. Conclusions: Only mild hypercoagulability was observed in patients who had undergone laparoscopic cholecystectomy. The cytokine surge was correlated with hypercoagulability. There was in fact a positive correlation between IL-6 level and hypercoagulability. The correlation between cytokine levels and coagulation activation may be related to the type of surgery performed. Further studies are required to investigate these issues. Presented at the annual meeting of the Society of American Gastrointestinal Endoscopic Surgeons (SAGES), Los Angeles, CA, USA, 10–15 March 2003     

Th1-like and Th2-like cytokines in patients undergoing open versus laparascopic cholecystectomy

The advantages of laparoscopic (LC versus, open cholecystectomy (OC) seems to be related to minimal invasive procedure and to the moderate inflammatory response. The aim of this study is to define the involvement of Th1 (IFN-gamma) and Th2 (IL-4, IL-6, IL-10, IL-13) cytokines production in vivo and in vitro in patients undergoing OC or LC. In 42 patients undergoing LC (n = 22) and OC (n = 20) Th1-like and Th2-like was evaluated before operation and at 6, 24 and 48 hours after operation for white blood cell counting and cytokines (IL-4, IL-6, IL-10, IL-13, IFN-gamma, TNF-alpha) in the sera and in the supernatants from circulating mononuclear cells stimulated with phytohemagglutinin or lipopolysaccharide. The acute phase response cytokine, IL-6, appeared significantly increased following OC than after LC. All other cytokines did not very significantly. In vitro data shows a reduction of IFN-gamma and increase in Th2-like cytokines in OC patients compared with the basal value. In LC subjects we observed an high production of IFN-gamma associated to an increase of Th2-like cytokines, like IL-10 and IL-13, even though IL-4 and IL-6 were unmodified. In contrast to OC, LC did not significantly affect immunocompetence, maintaining a moderate inflammatory response and an adequate balance between Th1 and Th2 cytokine. Furthermore, the strong activation of cells producing Th1-like cytokines in LC patients following mitogen activation indicated a consistent anti-microbial activity, that was not detectable in OC patients, that showed after activation only a Th2 response.     

细胞因子对异种脱细胞真皮基质免疫调节作用的临床研究

目的　探讨烧伤患者接受异种 /异体脱细胞真皮基质 (xeno /allo ADM )移植后全身和局部多种细胞因子与移植物近期转归的关系。　方法　在大面积烧伤患者的四肢切痂创面上 ,移植xeno ADM (12例 ,19块 )或allo ADM(15例 ,18块 ) ,其上覆盖自体超薄断层皮片 ,并以 6例单纯移植自体中厚断层皮片 (auto TTS)的烧伤患者为对照。移植物成活后 4～ 8周 ,收集局部组织标本、血清和xeno ADM排斥后的创面渗出液 ,采用免疫组织化学染色与酶联免疫吸附法 (ELISA) ,检测白细胞介素 (IL) 1β、IL 4、IL 6、肿瘤坏死因子α(TNF α)和γ型干扰素 (IFN γ)的含量。　结果　免疫组织化学染色结果显示 ,移植物内IL 1β、IL 4、IL 6、TNF α和IFN γ的阳性细胞密度或着色强度相比较 ,xeno ADM >allo ADM >auto TTS (P <0 .0 5 )。ELISA检测结果显示 ,xeno ADM被排斥后创面渗出液中IL 4、IL 6、TNF α和IFN γ水平明显高于自体血清 ,但其血清IL 4与IFN γ水平分别低于和高于未排斥时。xeno ADM移植后血清中IL 4、IFN γ水平明显高于allo ADM和auto TTS(P <0 .0 5～ 0 .0 1)。　结论　xeno ADM移植后可在局部检测到高水平的IL 1β、IL 4、IL 6、IFN γ ,可能与细胞杀伤和细胞因子介导的免疫放大作用有关。这些细胞因子的动态变     

Intestinal graft versus native liver cytokine expression in a rat model of intestinal transplantation with and without donor-specific cell augmentation

BACKGROUND: Immunomodulatory strategies such as donor-specific bone marrow or blood transfusions have been used to promote engraftment after intestinal transplants. We previously showed that delivery of donor antigen via the portal vein can effectively reduce the rate of intestinal graft rejection. The purpose of our current study was to investigate the impact of donor-specific cell augmentation (blood versus bone marrow) via the portal vein on cytokine expression in intestinal grafts versus native livers. MATERIAL AND METHODS: We performed heterotopic small intestinal transplants between male Brown-Norway (donor) and female Lewis (recipient) rats. We studied 10 groups according to the type of donor-specific cell augmentation and the use and dose of immunosuppressive therapy. For cell augmentation, donor-specific blood or bone marrow was transfused via the donor portal vein immediately before graft implantation. For immunosuppression, tacrolimus was used post-transplant at a high or low dose. Control rats received neither immunosuppression nor cell augmentation. Tissue samples for histological assessment were obtained at designated time points. RNA was extracted from intestinal graft and native liver biopsies for cytokine measurements (IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IFN-gamma, TNF-alpha, and TNF-beta). Chimerism levels were determined using Q-PCR analysis. RESULTS: Without concurrent immunosuppression, neither portal donor-specific blood nor bone marrow transfusion reduced the rate of rejection. With immunosuppression, outcome was significantly better after portal donor-specific blood (versus bone marrow) transfusion. Irrespective of the type of donor-specific cell augmentation, severe rejection caused strong cytokine expression in the grafts of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha; in the native livers, mainly of TNF-alpha (with IFN-gamma showing hardly any increase). In general, rejection caused stronger cytokine expression in the grafts than in the native livers. Mild rejection correlated well with strong intragraft expression of IL-6, TNF-alpha, and TNF-beta (early rejection markers); severe rejection with IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha (late rejection markers).In addition to cell augmentation per se, the type of cell augmentation also had an impact on cytokine expression in both grafts and native livers. Cell-augmented (versus tacrolimus-treated) rats showed hardly any differences in intragraft cytokine expression, but the expression of almost all cytokines was significantly stronger in the native livers. With immunosuppression, bone marrow infusion increased intragraft cytokine expression of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF alpha, as well as liver cytokine expression of IL-1 beta, compared to blood transfusion. This finding reflected the more advanced rejection stages in the bone marrow infused group; different types of donor-specific cell augmentation had similar effects on liver cytokine expression. In the absence of myoablative therapy, chimerism levels were low, in both cell-augmented and non-cell-augmented groups. CONCLUSIONS: Rejection and donor-specific cell augmentation independently causes differences in intragraft versus native liver cytokine expression after intestinal transplants. Portal donor-specific blood transfusion, as compared with donor-specific bone marrow infusion, lowered the incidence of rejection and diminished intragraft cytokine up-regulation.     

Effect of CO2 pneumoperitoneum on the systemic and peritoneal cytokine response in a LPS-induced sepsis model

We studied the effect of carbon dioxide (CO2) pneumoperitoneum on the systemic and peritoneal cytokine response in a rat model of intraperitoneal sepsis. After intraperitoneal injection of bacterial lipopolysaccharide (LPS, 10 mg/kg), rats were divided into 3 groups (n = 49 in each group): control (abdominal puncture); CO2 pneumoperitoneum, and laparotomy. Blood and peritoneal lavage fluid (PLF) were sampled at 0, 1, 2, 3, 4, 6, and 8 h after LPS challenge. Blood cell counts, plasma endotoxin level, and the levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) in the plasma and PLF were measured. Blood cell counts did not differ between the 3 groups. Plasma endotoxin levels in the pneumoperitoneum group were significantly increased immediately after the procedure (p < 0.05). Although peak plasma TNF-alpha levels in the pneumoperitoneum group were seen immediately after the procedure, other changes in plasma cytokine levels did not differ significantly between the 3 groups. PLF TNF-alpha and IL-1beta levels in the pneumoperitoneum group were significantly lower than levels in the control and laparotomy groups soon after the procedure (p < 0.05). PLF IL-6 levels in the pneumoperitoneum group tended to be lower than those in the laparotomy group. In conclusion, CO2 pneumoperitoneum might induce different responses between systemic and peritoneal cytokines soon after the procedure in a rat model of intraperitoneal sepsis.