| Objective: To investigate the gut barrier dysfunction during intra-abodominal infection and manifest the protect effect of QingReJieDu Herbs. To investigate whether immune disturbance occurs during intra-abodominal infection both in blood and mesenteric lymph node (MLN), study the role of Treg in mechanism of immune imbalance, manifest the effect of QRJD herbs on ameliorating the systemic immune balance. To test the hypothesis that Immune dysfunction of T cells in MLN may aggravate intestinal endotoxin translocation which correlated to systemic immune imbalance.Methods: The rat model of multiple organ dysfunction syndrome was prepared by intra-abdominal E. coli administration. The rats were divided 3 groups randomly, including the normal control group (NOR), model group (CON), and QRJD group. 6h, 24h and 72h after the administration, the histomorphology of ileum was observed, the level of plasma endotoxin, serum cytokines, ALT, Cr, and MPO of the lung tissue were determined, the L/M ratio in urine was detected. The FITC-LPS was poured down the stomach as the tracer to investigate its concentration in the portal vein blood, the lymph in the thoracic duct, the arterial blood and the tissues samples of the mesenteric lymph node (MLN), lung, liver, kidney, intestine, spleen. The effects of QRJD herbs on these indicators were also investigated. 6h, 12h, 24h, 48h and 72h after the bacterial administration, samples of abdominal aorta and MLN were collected, Th1 :Th2 ratio and Treg percentage of CD4~+ T cell were determined by means of flow cytometer.Results: The intestinal barrier histomorphology and many important internal organs appeared obvious pathological lesions, the level of plasma endotoxin, TNF-α, IL-6, IL-10, L/M ratio, and biochemical indicators were increased in model group during intra-abodominal infection. The indicators of the QRJD group were better than those of the model group. The FITC-LPS concentration is higher in MLN, lung, liver and thoracic duct than other tissues. The level of Th1:Th2 ratio in blood was elevated in early period of infection(6h and 12h), but lowered in late period of sepsis(from 24h to 72h) in model group, QRJD herbs could decrease the level of Th1/Th2 in early period and increase it in late period. Treg percentage of CD4~+ T cell in blood was decreased 6h after model was reproduced, and then was increasing during the end of the observation in the model group. QRJD herbs could lower the elevation of its percentage during the late phase. The level of Th1:Th2 ratio in MLN was elevated in early period of infection(6h), then dramatically decreased subsequently in model group, QRJD herbs could decrease the improvement of Thl/Th2 in early period and increase it in late period. Treg percentage of CD4~+ T cell in MLN was decreased 6h after model was reproduced, and then was increasing during next periods in the model group. QRJD herbs could lower the elevation of Treg percentage during the late phase.Conclusion: The animal model of MODS was established successfully by this method. Intra-abodominal infection can lead to the damage of gut barrier function, translocation of intestinal endotoxin, release of cytokines. QRJD herbs could protect the gut barrier function, significantly inhibit the translocation of gut origin endotoxin and also could significantly decrease the level of serum TNF-α, IL-6, IL-10 in peritonitis rats. Gut origin endotoxin could mainly be absorbed into the circulatory system via mesenterium lymph system during the early phase of intra-abodominal infection. There is a dissonance of immunological function both in systemic circulation and MLN during intra-abdominal infection, showing a depression in T cell immunological function, whereas Treg may be one of the reasons related to sepsis-induced immunoparalysis. Immune dysfunction of T cells in MLN may aggravate intestinal endotoxin translocation which correlated to systemic immune imbalance. QRJD herbs can obviously ameliorate the immunological disturbance in sepsis in acute peritonitis rats. |
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