南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (05): 625-.

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日本血吸虫半胱氨酸蛋白酶抑制剂干预脂多糖诱导的小鼠脓毒症

万勇坤，李徽徽，左琳，王小莉，王黎源，贺文欣，姜辉，王守祥，盛洁，张敏，钱海春，杨芳芳，谢红， 高世芳，方强，杨小迪，刘牧林   

  • 出版日期:2018-05-20 发布日期:2018-05-20

Intervention with Schistosoma japonicum cysteine protease inhibitor for treatment of lipopolysaccharide-induced sepsis in mice

  • Online:2018-05-20 Published:2018-05-20

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摘要： 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。

Abstract: 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.