ABSTRACT
Objective:To explore the mechanism of complement 5a (C5a) in the pathogenesis of sepsis.Methods:SPF male C57BL/6J mice were selected and divided into sham operation group (Sham group), cecal ligation and puncture (CLP) group and CLP+anti-C5A monoclonal antibody intervention group (CLP+anti-C5a group) according to random number table with 20 mice in each group. A CLP model was reproduced to induce sepsis, and those in the Sham group only underwent laparotomy without ligation and perforation. In the CLP+anti-C5a group, 0.15 mg of anti-C5a monoclonal antibody was injected intraperitoneally immediately after CLP, and in the Sham group and CLP group were given equal amount of normal saline. The cumulative survival rate was analyzed by Kaplan-Meier method. Serum levels of tumor necrosis factor-α (TNF-α), interleukins (IL-12, IL-4), and interferon-γ (IFN-γ) were measured 24, 48 and 72 hours after operation by enzyme linked immunosorbent assay (ELISA). Immunohistochemical staining was used to observe the expression of C5a receptor (C5aR) in lung and kidney tissues 48 hours after operation. The proportions of dendritic cell (DC), regulatory T cell (Treg) and helper T cell 17 (Th17) in splenic mononuclear cells 48 hours after operation were analyzed by flow cytometry.Results:The 7-day cumulative survival rate of mice in the CLP group was significantly lower than that in the Sham group (30.00% vs. 100.00%; Log-Rank test: χ 2 = 47.470, P < 0.001), and the peripheral blood inflammatory mediators TNF-α, IL-12 and IL-4 were increased 24 hours after operation, followed by a significant decreasing at 48 hours, and then gradually increased at 72 hours. IFN-γ gradually increased 24 hours after operation and lasted for 72 hours. Immunohistochemistry showed that a large number of C5aR was expressed in pulmonary and renal endothelial cells 48 hours after operation in the CLP group. Compared with the Sham group, the proportion of DC [(1.80±0.30)% vs. (6.90±1.20)%, P < 0.05] and Treg [(0.38±0.02)% vs. (4.00±0.50)%, P < 0.05] in splenic mononuclear cells was down-regulated in the CLP group, the proportion of Th17 was up-regulated [(0.83±0.08)% vs. (0.32±0.03)%, P < 0.05], and disorder of immune function was found. After anti-C5A monoclonal antibody intervention, the 7-day cumulative survival rate increased significantly compared with the CLP group (54.54% vs. 30.00%; Log-Rank test: χ 2 = 28.090, P < 0.001); TNF-α, IL-12 and IFN-γ were further increased, while IL-4 was significantly decreased; the expression of C5aR in lung and kidney tissues were significantly decreased, and the expression of mature DC cells [(5.10±1.20)% vs. (1.80±0.30)%, P < 0.05] and Treg [(2.58±0.05)% vs. (0.38±0.02)%, P < 0.05] in spleen were significantly increased compared with the CLP group, and Th17 was significantly decreased [(0.54±0.05)% vs. (0.83±0.08)%, P < 0.05]. Conclusion:It is preliminarily concluded that anti-C5A monoclonal antibody may improve the prognosis of sepsis by improving the polarization of mature DC and T cells in the spleen, and C5a plays an important role in the immune regulation of sepsis cells.
ABSTRACT
Objective To investigate the therapeutic effect of Radish Seed Chengqi Decoction combined with glutamine for treatment of patients with sepsis complicated with paralytic ileus. Methods Sixty patients with septic ileus admitted to the Department of Emergency Intensive Care Unit (EICU) of Tianjin Medical University General Hospital from August 2017 to January 2019 were enrolled. The Western medicine treatment group (30 cases) received routine Western medical methods including intravenous drip of glutamine, etc. for symptomatic treatment, and the Radish Seed Chengqi Decoction group (30 cases), based on the treatment in the Western medicine group, were additionally given the decoction by nasal feeding and retention enema twice a day, once 100 mL with the course of 7 days in both groups. The herbal prescription of Radish Seed Chengqi Decoction contained rhubarb 24 g (decocted later), sodium sulfate 15 g (mixed with water to take), magnoliae cortex 15 g, orange frui 10 g, aucklandiae 10 g, henan achyranthes root 10 g and radish seed 15 g, constituting one dose, two doses one day given to a patient, making one dose of herbal medicine to 200 mL of decoction for nasal feeding and retention enema, once 100 mL respectively, twice a day for 7 days. After treatment, the changes of survival rate in ICU, the total effective rate and the level of procalcitonin (PCT) were observed in both groups. Results After treatment, the total effective rate in Radish Seed Chengqi Decoction group was significantly higher than that in the Western medicine treatment group [83.33% (25/30) vs. 66.67% (20/30), P < 0.05], and the ICU 7-day survival rate in Radish Seed Chengqi Decoction group was also significantly higher than that in Western medicine treatment group [90.00% (27/30) vs. 76.67% (23/30), P < 0.05]. After treatment, the levels of PCT (μg/L) in both groups were significantly decreased compared with those before the treatment (Western medicine treatment group: 3.38±1.72 vs. 6.43±3.29, and Radish Seed Chengqi Decoction group: 2.39±1.24 vs. 6.78±2.31, both P < 0.05), and the degree of decrease of PCT in radish seed chengqi decoction group was more obvious than that in Western medicine treatment group (μg/L: 2.39±1.24 vs. 3.38±1.72, P < 0.05). Conclusion Applying alanyl glutamine combined with Radish Seed Chengqi Decoction for treatment of patients with sepsis complicated with paralytic ileus can significantly improve the clinical symptoms, reduce the PCT level and elevate the survival rate of patients.