本文選題：吳茱萸堿　切入點(diǎn)：急性腹膜炎　出處：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2017年02期

【摘要】：目的探討吳茱萸堿(evodiamine,EVO)對酵母多糖(zymosan,ZYM)誘導(dǎo)的小鼠腹膜炎中炎癥反應(yīng)及中性粒細(xì)胞(polymorphonuclear neutrophil,PMN)浸潤的影響。方法將54只成年雄性C57BL/6小鼠按隨機(jī)數(shù)字表法分為正常對照組、模型組和EVO治療組。模型組腹腔注射1 mg/m L ZYM溶液(1 m L)建立急性腹膜炎小鼠模型,EVO治療組則同時(shí)腹腔注射10 mg/kg EVO及1 mg/m L ZYM溶液,正常對照組腹腔注射等量磷酸鹽緩沖液。各組處理后2、6、12 h處死6只小鼠,收集眼眶血及腹腔灌洗液。采用酶聯(lián)免疫吸附法檢測血清及腹腔灌洗液中白介素6(interleukin-6,IL-6)、腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)、角化細(xì)胞來源趨化因子(keratinocyte-derived chemokine,KC)及巨噬細(xì)胞炎性蛋白2(macrophage inflammatory protein 2,MIP-2)的含量;通過血細(xì)胞計(jì)數(shù)板計(jì)數(shù)腹腔灌洗液中混合細(xì)胞總數(shù);利用流式細(xì)胞儀檢測腹腔灌洗液中PMN比例及PMN中活性氧(reactive oxygen species,ROS)水平;采用Western blot檢測6 h時(shí)腹腔灌洗液混合細(xì)胞中核轉(zhuǎn)錄因子-k B(nuclear factor-κB,NF-κB)p65入核情況。結(jié)果建模后2、6、12 h,模型組血清及腹腔灌洗液中IL-6、TNF-α、KC及MIP-2含量均高于正常對照組(P0.05),EVO治療組上述指標(biāo)均較模型組明顯降低(P0.05),其中,血清中IL-6、KC及MIP-2均于2 h達(dá)高峰,隨后逐漸下降,而TNF-α在2 h逐漸上升,6 h達(dá)到高峰,隨后逐漸下降;腹腔灌洗液中TNF-α、KC及MIP-2均于2 h達(dá)高峰,隨后逐漸下降,而IL-6在2 h逐漸上升,6 h達(dá)到高峰,隨后逐漸下降。制模后6 h,EVO治療組可顯著降低腹腔混合細(xì)胞數(shù)與PMN數(shù)及其百分比(P0.05),且PMN中ROS顯著降低(P0.05)。另EVO治療可顯著降低腹腔混合細(xì)胞中細(xì)胞核的NF-κB p65表達(dá)(P0.05)。結(jié)論 EVO可抑制ZYM誘導(dǎo)急性小鼠腹膜炎模型中血清及腹腔灌洗液中炎癥因子及趨化因子的分泌,減輕腹腔PMN的浸潤,并抑制PMN中ROS的產(chǎn)生,該作用可能是通過抑制NF-k B p65入核實(shí)現(xiàn)的。
[Abstract]:Objective to investigate the effects of evodiamine EVOO on inflammatory response and polymorphonuclear neutrophil PMNs infiltration in mice peritonitis induced by yeast polysaccharide zymosanine. Methods 54 adult male C57BL/6 mice were randomly divided into normal control group. Model group and EVO group. The model group was treated by intraperitoneal injection of 1 mg/m L ZYM solution (1 mL) to establish acute peritonitis mouse model. The model group was treated with 10 mg/kg EVO and 1 mg/m L ZYM solution at the same time. In the normal control group, the same amount of phosphate buffer solution was injected intraperitoneally. Six mice were killed at 12 h after treatment in each group. The contents of interleukin-6 interleukin-6 (IL-6), tumor necrosis factor- 偽 (TNF- 偽), keratinocyte derived chemokine (KC) and macrophage inflammatory protein (2(macrophage inflammatory protein 2MIP-2) in serum and peritoneal lavage fluid were detected by Elisa. The total number of mixed cells in peritoneal lavage fluid was counted by blood cell counter, the ratio of PMN in peritoneal lavage fluid and reactive oxygen speciesrose in PMN were measured by flow cytometry. The nuclear transcription factor -k B(nuclear factor- 魏 B, NF- 魏 B)p65 in peritoneal lavage fluid mixed cells was detected by Western blot at 6 h. Results the levels of IL-6 TNF- 偽 KC and MIP-2 in serum and peritoneal lavage fluid in model group were higher than those in normal control group (P0.05EVO) at 12 h after modeling. Compared with the model group, the indexes were significantly lower than that of the model group. Serum IL-6 KC and MIP-2 reached their peak at 2 h, then decreased gradually, while TNF- 偽 gradually increased at 2 h and reached the peak at 6 h, then decreased gradually, and TNF- 偽 KC and MIP-2 in peritoneal lavage fluid reached the peak at 2 h and then decreased gradually. However, IL-6 gradually increased at 2 h and reached the peak at 6 h. After 6 h of model making, the number of mixed cells and the number and percentage of PMN in abdominal cavity were significantly decreased in EVO group, and the ROS in PMN decreased significantly (P 0.05). In addition, the expression of NF- 魏 B p65 in the nucleus of peritoneal mixed cells was significantly decreased by EVO treatment. Conclusion EVO can inhibit the secretion of inflammatory factors and chemokines in serum and peritoneal lavage fluid of acute peritonitis induced by ZYM in mice. It can reduce the infiltration of PMN in abdominal cavity and inhibit the production of ROS in PMN, which may be achieved by inhibiting the entry of NF-k B p65 into the nucleus.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第二醫(yī)院肝膽外科;第三軍醫(yī)大學(xué)大坪醫(yī)院野戰(zhàn)外科研究所第一研究室 創(chuàng)傷、燒傷與復(fù)合傷國家重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(81372088)~~
【分類號】：R259

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