傅泳,刘颖,刘曼,等.海兔素对酒精诱导大鼠肠上皮细胞屏障损伤的保护作用[J].中国海洋药物,2016,35(2):65-71.
海兔素对酒精诱导大鼠肠上皮细胞屏障损伤的保护作用
The protective effects of Aplysin on alcoholic induced barrier injury of intestinal epithelial cells in rats
投稿时间:2015-08-12  修订日期:2015-10-16
DOI:
中文关键词:  海兔素  酒精性肠粘膜损伤  TNF-α  TGF-β  FABP2  FOX04
English Keywords:Aplysin  Alcoholic induced intestinal mucosal damage  FABP2  TNF-α  TGF-β  FOX04
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作者单位E-mail
傅泳 青岛大学 fuyongfelcia@163.com 
刘颖 青岛大学  
刘曼 青岛大学  
王文成 青岛大学  
苏彬 青岛大学  
梁惠* 青岛大学 qdlianghui@126.com 
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中文摘要:
      目的 探讨海兔素(Aplysin)对酒精诱导大鼠肠上皮细胞屏障损伤的保护作用。方法 60只Wistar大鼠随机分为4组,空白对照组给予10 mL/(kg?bw?d)生理盐水灌胃;酒精模型组给予56 ℃红星二锅头灌胃,第1周5.5 mL/(kg?bw?d),第2周7.5 mL/(kg?bw?d),第3周8.0 ml/ (kg?bw?d),第4周9.0 ml/(kg?bw?d),第5周至第8周11 ml/kg/d;海兔素低、高剂量组分别给予Aplysin100、150 mg/(kg?bw?d)灌胃,酒精剂量同模型组,持续8周。末次灌胃后禁食12h,腹主动脉取血及小肠组织。透射电镜观察小肠超微结构变化;酶联免疫吸附实验(ELISA)检测大鼠血浆中FABP2、TNF-α和TGF-β浓度;免疫组化实验检测小肠组织中FOXO4蛋白的表达情况。结果 电镜观察发现,空白对照组小肠上皮柱状细胞排列整齐、紧密,微绒毛丰富,紧密连接结构清晰完整;酒精模型组小肠微绒毛排列稀疏,紧密连接肿大;海兔素组较酒精模型组得到不同程度改善。空白对照组血浆FABP2、TNF-α和TGF-β浓度分别为(2.79±1.18)ng/mL、 (30.42±18.89)pg/mL和 (38.67±19.22)pg/mL,酒精模型组与其相比显著升高,分别达到(3.87±1.01)ng/mL、(59.31±35.01)pg/mL、和(62.28±9.33)pg/mL;海兔素低剂量组分别为(2.87±1.60)ng/mL 、(36.32±19.58)pg/mL、和(42.09±19.60)pg/mL,高剂量组为(2.25±1.06)ng/mL、(32.50±18.70)pg/mL和 (33.91±22.54)pg/mL,均较酒精模型组明显降低。酒精模型组小肠组织FOXO4表达水平明显降低,OD值为(0.115±0.064) ,与正常对照组(OD =0.264±0.023)比较,具有显著性差异(P<0.05);海兔素低、高剂量组FOXO4的OD值分别为(0.201±0.089)、(0.231±0.092),均高于酒精模型组(P<0.05)。结论 海兔素对酒精诱导的大鼠肠上皮细胞屏障损伤具有一定保护作用,其作用机制可能与海兔素调节炎性反应相关蛋白表达,减少炎性因子分泌有关。
English Summary:
      Abstract: Objective To observe the protective effects of Aplysin on alcoholic induced barrier injury of intestinal epithelial cells in rats Methods Male Wistar rats were randomly divided into the following four groups: control group, normal diet with normal saline,ethanol-treated group, normal diet with ethanol administration,and low- and high-dose Aplysin[ 100, and 150 mg/(kg?bw?d)]plus ethanol treatment groups. Excluding the rats in the normal control group, the other animals were initially received intragastric administration with 56% (v/v) ethanol 5.5 mL kg-1day-1 for the first week , 7.5 mL kg-1day-1 for the secend week , 8 mL kg-1day-1 for the third week,9 mL kg-1day-1 for the fourth week ,and 12 mL kg-1day-1for the rest of the 4 weeks. Twelve hours after the last ethanol treatment, the rats were anesthetized, and blood was collected, the small intestine was obtained for the observation of ultrastructural changes by tansmission electron microscope; The expression of FOX04 in small intestine was detected by immunohistochemistry; plasma FABP2 TGF-β and TNF-α concentration was measured by enzyme linked immunosorbent assay. Results in the ethanol-treated group, the ultrastructural changes of small intestine tissue by tansmission electron microscope showed that the intestinal villi fell away, and loosely arranged, tight junction were swelling, however, the control group was not show abnormal ultrastructural.. and in each dose Aplysin, ultrastructural changes were better than the ethanol-treated group. In control group, the plasma concentration of FABP2 , TNF-α and TGF-βwere (2.79±1.18)ng/ml, (30.42±18.89)pg/ml ang (38.67±19.22)pg/ml, the ethanol-treated group were (3.87±1.01)ng/ml, (59.31±35.01)pg/ml and (62.28±9.33)pg/ml, it was significantly elevated compared with control group. the plasma concentration of FABP2 , TNF-α and TGF-β in alcohol aplysin (100mg/kg) group were (2.87±1.60)ng/ml, (36.32±19.58)pg/ml and (42.09±19.60)pg/ml, alcohol aplysin (150mg/kg) group were (2.25±1.06)ng/ml, (32.50±18.70)pg/ml and (33.91±22.54)pg/ml, they were significantly reduced compared with the ethanol-treated group. The fox04 expression in Ethanol-treated group was decreased with comparison of control group, OD value in Ethanol-treated group and control group were (0.115±0.064) and (0.264±0.023);compared with the ethanol-treated group , in each dose Aplysin the expression of FOX04 were increased(P<0.05), alcohol aplysin (100 and 150mg/kg) group were (0.201±0.089)、(0.231±0.092). Conclusion use of Aplysin shows some protective effect on intestinal mucosal damage induced by alcohol exposure in rats. Its mechanism may be associated with Aplysin regulate the expression of Inflammatory reaction related proteins, and reduce the release of inflammatory factor.
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