本文選題：旋毛蟲　切入點：膿毒癥　出處：《中國寄生蟲學與寄生蟲病雜志》2017年02期 　論文類型：期刊論文

【摘要】：目的觀察旋毛蟲(Trichinella spiralis)成蟲排泄-分泌蛋白(adult-worm excretory-secretory protein,AES)對小鼠膿毒癥急性肝損傷的保護作用。方法 24只雄性BALB/c小鼠采用抽簽法隨機分為假手術組(Sham組)、盲腸結扎穿孔組(CLP組)、AES治療組(AES+CLP組)和AES對照組(AES+Sham組),每組6只。CLP組和AES+CLP組采用盲腸結扎穿孔術制備膿毒癥急性肝損傷動物模型,Sham組和AES+Sham組不結扎、不穿孔盲腸,其余操作與CLP組相同。術后30 min,Sham組和CLP組腹腔注射PBS,AES+CLP組和AES+Sham組腹腔注射AES(25μg/只)。術后12 h,檢測小鼠血清中丙氨酸轉氨酶(ALT)和天門冬氨酸轉氨酶(AST)水平,觀察小鼠肝臟組織病理改變,蛋白質印跡(Western blotting)測定肝臟組織中Toll樣受體4(TLR4)、髓樣分化因子88(My D88)的表達水平,免疫組化法測定肝細胞核因子-κB(NF-κB)p65的表達水平并分析NF-κB p65核陽性率,ELISA檢測小鼠肝臟組織中α腫瘤壞死因子(TNF-α)、白細胞介素-6(IL-6)和IL-1β的水平。結果術后12 h,CLP組小鼠血清中ALT和AST水平分別為(269.83±77.60)和(712.00±186.52)IU/L,明顯高于Sham組的(45.00±15.01)和(132.50±36.95)IU/L,及AES+CLP組的(136.67±30.27)和(419.00±60.86)IU/L(P0.05);AES+Sham組分別為(50.83±13.24)和(134.67±35.78)IU/L,與Sham組比較差異無統(tǒng)計學意義(P0.05)。HE染色顯示:CLP組肝索紊亂,肝細胞水腫,有炎性細胞浸潤;AES+CLP組肝臟結構損傷明顯減輕。Western blotting檢測顯示:CLP組小鼠術后肝臟組織中TLR4/β-actin和My D88/β-actin相對表達量分別為0.66±0.12和0.69±0.13,明顯高于Sham組的0.31±0.06和0.22±0.04(P0.05);AES+CLP組TLR4/β-actin相對表達量為0.56±0.09,與CLP組的比較差異無統(tǒng)計學意義(P0.05),而My D88/β-actin相對表達量為0.47±0.06,較CLP組降低(P0.05)。免疫組化分析結果顯示:CLP組肝細胞中NF-κB p65核陽性率為(18.93±2.91)%,明顯高于Sham組的(0.50±0.21)%及AES+CLP組的(9.60±1.59)%(P0.05)。ELISA檢測結果顯示:CLP組小鼠肝臟組織中TNF-α、IL-6和IL-1β水平分別是(26.21±2.96)、(80.80±9.94)和(28.99±3.70)pg/mg,明顯高于Sham組的(15.42±1.21)、(47.70±4.77)和(16.86±1.68)pg/mg,及AES+CLP組的(18.93±2.04)、(61.82±7.24)和(23.18±2.21)pg/mg(P0.05)。結論AES可通過降低肝臟組織中My D88表達和NF-κB p65核陽性率,從而降低肝臟組織中TNF-α、IL-6和IL-1β水平,減輕膿毒癥對肝臟的損傷。
[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 偽-tumor necrosis factor TNF- 偽, interleukin-6- (IL-6) and IL-1 尾 in the liver tissue of mice. Results 12 hours after operation, the serum levels of 偽-tumor necrosis factor TNF- 偽, interleukin-6 and IL-1 尾 were detected in mice in CLP group. The levels of ALT and AST were 269.83 鹵77.60) and 712.00 鹵186.52, respectively, which were significantly higher than those of Sham group (45.00 鹵15.01) and AES CLP group (132.50 鹵36.95) and AES CLP group (136.67 鹵30.27) and 419.00 鹵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 鹵0.12 and 0.69 鹵0.13, respectively, which were significantly higher than those in the Sham group (0.31 鹵0.06 and 0.22 鹵0.04P0.05AES / 尾 -actin). Compared with CLP group, the relative expression of my D88 / 尾 -actin was 0.47 鹵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 鹵2.91% in Sham group, which was significantly higher than that in Sham group (0.50 鹵0.21%) and AES CLP group. The results of TNF- 偽 IL-6 and IL-1 尾 were 26.21 鹵2.96, 80.80 鹵9.94 and 28.99 鹵3.70 PG / mg, respectively, which were significantly higher than those in Sham group (15.42 鹵1.21 鹵4.77) and 16.86 鹵1.68 PG / mg, and in AES CLP group (61.82 鹵7.24) and 23.18 鹵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- 偽, IL-6 and IL-1 尾 in liver tissue were decreased, and the liver injury caused by sepsis was alleviated.
【作者單位】： 蚌埠醫(yī)學院基礎醫(yī)學院;蚌埠醫(yī)學院安徽省感染與免疫重點實驗室;蚌埠醫(yī)學院第一附屬醫(yī)院;蚌埠醫(yī)學院第二附屬醫(yī)院;
【基金】：國家自然科學基金(No.81441120) 高�？蒲袆�(chuàng)新平臺團隊項目(No.2016-40) 安徽省自然科學基金(No.1508085QH158) 安徽省教育廳資助項目(No.KJ2015B013,KJ2017A235,KJ2017A236) 安徽省高校優(yōu)秀青年人才支持計劃重點項目(No.gxyq ZD2016159) 國家大學生創(chuàng)新創(chuàng)業(yè)訓練計劃項目(No.201610367009)~~
【分類號】：R459.7

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