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目的探讨旋毛虫成虫排泄分泌蛋白(AES)对盲肠结扎穿孔术(CLP)诱导的小鼠脓毒症的抑制作用。方法将48只BALB/c小鼠随机分为3组,即假手术对照组(PBS+sham组,A组)、脓毒症组(PBS+CLP组,B组)、AES蛋白治疗组(AES+CLP组,C组)。A组和B组于CLP术后经腹腔注射200μl PBS,C组术后经腹腔注射含25μg AES蛋白的等量PBS。每组随机抽取8只,建模后观察各组小鼠的一般情况及96 h生存率;每组其余8只小鼠于建模后12 h取肝、肺、肾,HE染色观察病理改变;以酶联免疫吸附试验(ELISA)检测小鼠血清中TNF-α、IL-1β、IL-6、IL-10及TGF-β细胞因子水平的变化。结果 3组小鼠96 h生存率差异有统计学意义(χ~2=21.16,P<0.05);与A组(100%)相比,B组小鼠生存率(0)降低(P<0.05),且肺、肝及肾脏损伤明显加重;经AES治疗后,C组存活率(75%)明显增加(P<0.05),小鼠肺脏、肝脏及肾脏病理损伤亦显著缓解。3组小鼠血清中的促炎因子TNF-α(F=27.11,P<0.05)、IL-1β(F=18.75,P<0.05)及IL-6(F=100.93,P<0.05)水平差异均有统计学意义;B组小鼠的促炎因子水平均高于A组(P均<0.05);经AES治疗后,C组小鼠上述3种促炎因子水平较B组均显著下降(P均<0.05)。3组小鼠血清中的调节性细胞因子IL-10(F=10.88,P<0.05)及TGF-β(F=11.37,P<0.05)水平差异亦有统计学意义;经AES治疗后,C组小鼠IL-10及TGF-β分泌水平较B组亦明显增加(P<0.05)。结论旋毛虫AES对CLP诱导的BALB/c小鼠脓毒症有显著的缓解作用。
Objective To investigate the inhibitory effect of Trichinella spiralis adults excretory secretory protein (AES) on sepsis induced by cecal ligation and puncture (CLP) in mice. Methods 48 BALB / c mice were randomly divided into three groups: sham operation control group (PBS + sham group, group A), sepsis group (PBS + CLP group, group B) + CLP group, C group). Groups A and B were injected intraperitoneally with 200μl PBS after CLP. Group C received intraperitoneal injection of an equal volume of PBS containing 25μg AES protein. Eight rabbits were randomly selected from each group. After the model was established, the general condition and 96-h survival rate of mice in each group were observed. The other 8 mice in each group were harvested at the 12th day after modeling to observe the pathological changes of liver, lung and kidney. Serum levels of TNF-α, IL-1β, IL-6, IL-10 and TGF-β were measured by enzyme linked immunosorbent assay (ELISA). Results The 96-h survival rate was significantly different between the three groups (χ ~ 2 = 21.16, P <0.05). Compared with group A (100%), the survival rate ), And lung, liver and kidney injury significantly aggravated. After treatment with AES, the survival rate of group C (75%) increased significantly (P <0.05), and the pathological changes of lung, liver and kidney were also relieved. The difference of serum levels of proinflammatory cytokine TNF-α (F = 27.11, P <0.05), IL-1β (F = 18.75, P <0.05) and IL-6 (F = 100.93, P <0.05) The levels of proinflammatory cytokines in group B were significantly higher than those in group A (all P <0.05). After treatment with AES, the levels of these three proinflammatory cytokines in group C were significantly lower than those in group B P <0.05). The levels of regulatory cytokines IL-10 (F = 10.88, P <0.05) and TGF-β (F = 11.37, P <0.05) in the three groups were also significantly different. Compared with group B, the secretion of IL-10 and TGF-β in mice also increased significantly (P <0.05). Conclusion Trichinella spiralis AES can relieve CLP-induced sepsis in BALB / c mice.