南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (4): 514-520.doi: 10.12122/j.issn.1673-4254.2021.04.05

• • 上一篇    下一篇

五步蛇毒蛋白C激活剂在脓毒症大鼠早期适应性免疫应答中的作用

孙 瑶，包鹏举，张根葆，王海华，王国栋   

  • 出版日期:2021-04-20 发布日期:2021-04-30

Effect of protein C activator from Agkistrodon acutus venom on early adaptive immune response of septic rats

  • Online:2021-04-20 Published:2021-04-30

RichHTML

51

PDF

354

摘要： 目的 探讨五步蛇毒蛋白C激活剂（PCA）在脓毒症大鼠早期适应性免疫应答中的作用。方法 采用SPF级SD大鼠78只，用随机数字表法分组：对照组（n=6，腹腔注射生理盐水）；通过腹腔注射脂多糖（LPS 10 mg/kg）复制脓毒症大鼠模型，模型组以LPS注射后4、6、8、12、16、24 h时的标本采集时间分为6组，6只/组；余36只大鼠于LPS注射30 min后使用药物干预，分为1-磷酸鞘氨醇受体1（S1PR1）激动剂SEW2871干预组（0.5 mg/kg，腹腔注射）和PCA干预组（0.1 mg/kg，腹腔注射），各干预组以LPS注射后6、12、24 h时的标本采集时间分为3组，每组6只。采用ELISA法检测血浆IL-4、S1P、IL-12和IFN-γ水平，应用免疫荧光法检测肠系膜淋巴结S1PR1和CD103表达的变化。结果 与对照组比较，脓毒症大鼠在注射LPS后的24 h内，血浆S1P、IL-12、IL-4和IFN-γ水平明显增高（P<0.05），LPS注射后6 h内IFN-γ/IL-4比值逐渐增高，之后逐渐降低；肠系膜淋巴结中S1PR1和CD103的表达明显增高（P<0.05）。SEW2871明显增加血浆S1P、IL-12和IFN-γ的浓度，降低血浆IL-4水平（P<0.05），且明显减少淋巴结中S1PR1和CD103的表达（P<0.05）。蛇毒PCA干预后，血浆IL-4水平明显增高，淋巴结S1PR1表达显著增多（P<0.05）。结论 五步蛇毒PCA对维持脓毒症早期机体炎症和免疫反应的平衡具有调节作用，其机制可能与S1P-S1PR1通路有关。

关键词:


Abstract: Objective To investigate the effect of protein C activator (PCA) from Agkistrodon acutus venom (AAV) in modulating early adaptive immune response of septic rats. Methods Rat models of sepsis were established by intraperitoneal injection of lipopolysaccharide (LPS; 10 mg/kg) in 36 SD rats, which were divided into 6 groups (n=6) for sample collection at 4, 6, 8, 12, 16 and 24 h after LPS injection, with 6 rats injected with saline as the control group. Another 36 rats were divided into two groups, and 30 min after LPS injection, the rats were treated with SEW2871 (a sphingosine- 1- phosphate receptor 1 agonist; 0.5 mg/kg) or PCA group (0.1 mg/kg), and each group was divided into 3 groups (n=6) for sample collection at 6, 12 and 24 h after LPS injection. Plasma IL-4, S1P, IL-12 and IFN-γ levels of the rats were detected using ELISA, and the expressions of S1PR1 and CD103 in the mesenteric lymph nodes were detected with immunofluorescence assay. Results The plasma levels of S1P, IL-12, IL-4 and IFN-γ (P<0.05) and the expressions of S1PR1 and CD103 in the mesenteric lymph nodes (P<0.05) all increased significantly in the rats 24 h after LPS injection; IFN-γ/IL-4 ratio increased progressively within 6 h after LPS injection and then subsided gradually. Compared with those in the corresponding sepsis model subgroups, the levels of S1P, IL-12 and IFN-γ increased while IL- 4 level decreased significantly (P<0.05), and the expression of S1PR1 and CD103 were reduced significantly (P<0.05) in SEW2871-treated rats; both the plasma level of IL-4 and the expression of S1PR1 in the mesenteric lymph nodes increased significantly in PCA-treated rats (P<0.05). Conclusion PCA can regulate the balance of inflammation and immune response in the early stage of sepsis in rats possibly through the S1P-S1PR1 pathway.

Key words:

acutus venom; protein C activator