Objective: To observe the effect of Schistosoma japonicum cysteine protease inhibitor (rSjCystatin) for treatment of lipopolysaccharide (LPS)-induced sepsis in mice.
Methods: After a week of adaptive feeding, 54 BALB/c mice were randomly divided into normal control group (group A), sepsis group (group B), and rSjCystatin intervention group (group C). The mice in group A received an intraperitoneal injection of PBS (100 µL), and those in groups B and C were injected with PBS (100 µL) containing LPS (10 mg/kg); the mice in group C were also intraperitoneally injected with 25 µg sjCystatin in 100 µL PBS 30 min after LPS injection. From each group, 10 mice were randomly selected 24 h after PBS or LPS injection for detecting serum levels of TNF-α, IL-6, and IL-10 using ELISA and the levels of ALT, AST, BUN, and Cr using automatic biochemical analyzer; the pathological changes in the liver, lung and kidney were observed with HE staining. The remaining 8 mice in each group were used for observing the changes in the general condition and the 72-h survival.
Results: The 72-h survival rates of the mice was 100% in group A, 0 in group B, and 36% in group C, showing a significant difference among the 3 groups (P<0.05). Compared with those in group A, the mice in group B exhibited obvious liver, lung, and renal pathologies with increased levels of ALT, AST, BUN, Cr, IL-6, and TNF-α (P<0.05). Treatment with sjCystatin significantly lessened LPS-induced organ pathologies, lowered the levels of liver and renal functional indexes and the pro-inflammatory cytokines, and increased the serum level of IL-10 in the mice (P<0.05).
Conclusion: SjCystatin can produce a significant therapeutic effect on sepsis induced by LPS in mice.
目的: 探讨日本血吸虫半胱氨酸蛋白酶抑制剂（SjCystatin）对脂多糖（LPS）诱导的小鼠脓毒症的干预效果。
方法: 54只BALB/c小鼠适应性饲养1周后，随机分为正常对照组(PBS组，A组)、脓毒症造模组（PBS+LPS组，B组）、蛋白干预组（PBS+ LPS+SjCystatin组，C组）。A组腹腔注射PBS（100 μL），B组、C组腹腔注射含LPS（10 mg/kg）的PBS（100 μL），其中C组小鼠于注射LPS后30 min腹腔注射含25 μg SjCystatin蛋白的PBS（100 μL）。每组随机抽取10只，造模后24 h取小鼠血清及肝、肺、肾组织，酶联免疫吸附试验（ELISA）验检血清中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、白细胞介素-10（IL-10）水平，全自动生化分析仪检测ALT、AST、BUN和Cr水平；肝、肺和肾组织HE染色观察其病理损伤；每组余8只小鼠，造模后记录小鼠72 h生存率的差别且观察小鼠状态的改变。
结果: 3组小鼠的72 h生存率具有统计学差异（P < 0.05），与A组（100%）相比，B组（0%）72 h生存率降低（P < 0.05），经过SjCystatin蛋白治疗后，C组生存率较B组增高（36%）。与A组相比，B组肝、肺、肾组织切片病理损伤加重，血清中ALT、AST、BUN、Cr、IL-6、TNF-α水平明显升高（P < 0.05）；与B组相比，C组调节因子IL-10水平明显升高，肝、肾、肺组织损伤明显减轻，且血清中上述肝肾功能检测指标及促炎因子水平明显降低（P < 0.05）。
结论: SjCystatin对LPS诱导的脓毒症有显著的治疗作用。