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

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. Keywords: sepsis; adaptive immune response; sphingosine-1-phosphate; sphingosine-1-phosphate receptor 1; Agkistrodon acutus venom; protein C activator
Effect of protein C activator from Agkistrodon acutus venom on early adaptive immune
response of septic rats
SUNห้องสมุดไป่ตู้Yao1, 2, BAO Pengju2, 3, ZHANG Genbao1, 2, WANG Haihua2, 4, WANG Guodong5 1Department of Pathophysiology, 2Institute of Snake Venom, 3School of Anesthesiology, 4Department of Physiology, 5School of Pharmacy, Wannan Medical College, Wuhu 241002, China
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、IL4 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：脓毒症；适应性免疫应答；1-磷酸鞘氨醇；1-磷酸鞘氨醇受体 1；五步蛇毒；蛋白 C 激活剂
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doi 10.12122/j.issn.1673-4254.2021.04.05
J South Med Univ, 2021, 41(4): 514-520
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 1，2，包鹏举 2，3，张根葆 1，2，王海华 2，4，王国栋 5 皖南医学院 1病理生理学教研室，2蛇毒蛇伤研究所，3麻醉学院，4生理学教研室，5药学院，安徽 芜湖 241002