旋毛虫成虫排泄-分泌蛋白对脓毒症急性肝损伤的保护作用及机制研究

发布时间：2018-03-21 19:34

本文选题：旋毛虫　切入点：脓毒症　出处：《中国寄生虫学与寄生虫病杂志》2017年02期 　论文类型：期刊论文

【摘要】：目的观察旋毛虫(Trichinella spiralis)成虫排泄-分泌蛋白(adult-worm excretory-secretory protein,AES)对小鼠脓毒症急性肝损伤的保护作用。方法 24只雄性BALB/c小鼠采用抽签法随机分为假手术组(Sham组)、盲肠结扎穿孔组(CLP组)、AES治疗组(AES+CLP组)和AES对照组(AES+Sham组),每组6只。CLP组和AES+CLP组采用盲肠结扎穿孔术制备脓毒症急性肝损伤动物模型,Sham组和AES+Sham组不结扎、不穿孔盲肠,其余操作与CLP组相同。术后30 min,Sham组和CLP组腹腔注射PBS,AES+CLP组和AES+Sham组腹腔注射AES(25μg/只)。术后12 h,检测小鼠血清中丙氨酸转氨酶(ALT)和天门冬氨酸转氨酶(AST)水平,观察小鼠肝脏组织病理改变,蛋白质印迹(Western blotting)测定肝脏组织中Toll样受体4(TLR4)、髓样分化因子88(My D88)的表达水平,免疫组化法测定肝细胞核因子-κB(NF-κB)p65的表达水平并分析NF-κB p65核阳性率,ELISA检测小鼠肝脏组织中α肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)和IL-1β的水平。结果术后12 h,CLP组小鼠血清中ALT和AST水平分别为(269.83±77.60)和(712.00±186.52)IU/L,明显高于Sham组的(45.00±15.01)和(132.50±36.95)IU/L,及AES+CLP组的(136.67±30.27)和(419.00±60.86)IU/L(P0.05);AES+Sham组分别为(50.83±13.24)和(134.67±35.78)IU/L,与Sham组比较差异无统计学意义(P0.05)。HE染色显示:CLP组肝索紊乱,肝细胞水肿,有炎性细胞浸润;AES+CLP组肝脏结构损伤明显减轻。Western blotting检测显示:CLP组小鼠术后肝脏组织中TLR4/β-actin和My D88/β-actin相对表达量分别为0.66±0.12和0.69±0.13,明显高于Sham组的0.31±0.06和0.22±0.04(P0.05);AES+CLP组TLR4/β-actin相对表达量为0.56±0.09,与CLP组的比较差异无统计学意义(P0.05),而My D88/β-actin相对表达量为0.47±0.06,较CLP组降低(P0.05)。免疫组化分析结果显示:CLP组肝细胞中NF-κB p65核阳性率为(18.93±2.91)%,明显高于Sham组的(0.50±0.21)%及AES+CLP组的(9.60±1.59)%(P0.05)。ELISA检测结果显示:CLP组小鼠肝脏组织中TNF-α、IL-6和IL-1β水平分别是(26.21±2.96)、(80.80±9.94)和(28.99±3.70)pg/mg,明显高于Sham组的(15.42±1.21)、(47.70±4.77)和(16.86±1.68)pg/mg,及AES+CLP组的(18.93±2.04)、(61.82±7.24)和(23.18±2.21)pg/mg(P0.05)。结论AES可通过降低肝脏组织中My D88表达和NF-κB p65核阳性率,从而降低肝脏组织中TNF-α、IL-6和IL-1β水平,减轻脓毒症对肝脏的损伤。
[Abstract]:Objective to observe the protective effect of Trichinella spiralis (Trichinella spiralis) on acute liver injury induced by sepsis in mice. Methods 24 male BALB/c mice were randomly divided into sham group and cecal ligation and perforation group by drawing lots. Six rats in each group and AES CLP group were treated with cecal ligation and perforation to make septic acute liver injury model. The animal model of acute hepatic sepsis injury was induced by cecal ligation and perforation. The rats in the sham group and AES Sham group were not ligated. The other operation was the same as that in the CLP group, the other operation was the same as that in the CLP group. 30 min after operation, the rats in the sham group and the CLP group were injected intraperitoneally with AES(25 渭 g / g in the CLP group and the AES Sham group. At 12 hours after operation, the serum levels of alanine aminotransferase (alt) and aspartate transaminase (AST) in the serum of the mice were measured. The expression of Toll like receptor 4 (TLR4) and myeloid differentiation factor (88(My D88) was determined by Western blotting. Immunohistochemical method was used to detect the expression of NF-魏 B nuclear factor-魏-魏 NF- 魏 B)p65 and to analyze the positive rate of NF- 魏 B p65. Elisa was used to detect the levels of 伪-tumor necrosis factor TNF- 伪, interleukin-6- (IL-6) and IL-1 尾 in the liver tissue of mice. Results 12 hours after operation, the serum levels of 伪-tumor necrosis factor TNF- 伪, interleukin-6 and IL-1 尾 were detected in mice in CLP group. The levels of ALT and AST were 269.83 卤77.60) and 712.00 卤186.52, respectively, which were significantly higher than those of Sham group (45.00 卤15.01) and AES CLP group (132.50 卤36.95) and AES CLP group (136.67 卤30.27) and 419.00 卤60.86LP0.05 Sham group respectively. Liver cell edema, The relative expressions of TLR4/ 尾 -actin and my D88 / 尾 -actin in the liver of mice in the CLP group were 0.66 卤0.12 and 0.69 卤0.13, respectively, which were significantly higher than those in the Sham group (0.31 卤0.06 and 0.22 卤0.04P0.05AES / 尾 -actin). Compared with CLP group, the relative expression of my D88 / 尾 -actin was 0.47 卤0.06, which was lower than that of CLP group. Immunohistochemical analysis showed that the positive rate of NF- 魏 B p65 was 18.93 卤2.91% in Sham group, which was significantly higher than that in Sham group (0.50 卤0.21%) and AES CLP group. The results of TNF- 伪 IL-6 and IL-1 尾 were 26.21 卤2.96, 80.80 卤9.94 and 28.99 卤3.70 PG / mg, respectively, which were significantly higher than those in Sham group (15.42 卤1.21 卤4.77) and 16.86 卤1.68 PG / mg, and in AES CLP group (61.82 卤7.24) and 23.18 卤2.21pggP0.05. conclusion AES can decrease the expression of my D88 and the positive rate of NFB p65 in liver tissue. Thus, the levels of TNF- 伪, IL-6 and IL-1 尾 in liver tissue were decreased, and the liver injury caused by sepsis was alleviated.
【作者单位】：蚌埠医学院基础医学院;蚌埠医学院安徽省感染与免疫重点实验室;蚌埠医学院第一附属医院;蚌埠医学院第二附属医院;
【基金】：国家自然科学基金(No.81441120) 高校科研创新平台团队项目(No.2016-40) 安徽省自然科学基金(No.1508085QH158) 安徽省教育厅资助项目(No.KJ2015B013,KJ2017A235,KJ2017A236) 安徽省高校优秀青年人才支持计划重点项目(No.gxyq ZD2016159) 国家大学生创新创业训练计划项目(No.201610367009)~~
【分类号】：R459.7

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