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

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10mL/(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9.0kg·bw·d),第5周至第8周11mL/(kg·bw·d);海兔素低、高剂量组分别给予Aplysin100、150mg/(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;免疫组化法测小肠组织中FOXO4表达情况。结果与空白对照组相比,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。与酒精模型组相比,空白对照组血浆FABP2、TNF-α和TGF-β浓度显著升高,差异具有显著性(P<0.05);而海兔素低、高剂量组均较酒精模型组明显降低(P<0.05)。酒精模型组小肠组织FOXO4表达水平明显降低,与正常对照组比较,具有显著性差异(P<0.05);海兔素低、高剂量组FOXO4表达水平较酒精模型组显著升高(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。

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 150mg/（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 5.5mL·kg^-1d^-1 for the first week,7.5mL·kg^-1d^-1 for the second week,8mL·kg^-1d^-1 for the third week,9mL·kg^-1d^-1 for the fourth week,and 12mL·kg^-1d^-1 for the rest of the 4weeks.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 FOXO4 in small intestine was detected by immunohistochemistry;plasma FABP2TGF-βand TNF-αconcentration 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 did not show abnormal ultrastructural.In each dose Aplysin,ultrastructural changes were better than the ethanol-treated group.The plasma concentration of FABP2,TNF-αand TGF-βin the ethanol-treated group were significantly elevated compared with control group.But the plasma concentration of FABP2,TNF-αand TGF-βin alcohol＋ Aplysin（100mg/kg and 150mg/kg）group were significantly reduced compared with the ethanol-treated group.The FOXO4 expression in ethanol-treated group was decreased with comparison of control group;compared with the ethanol-treated group,in each dose Aplysin the expression of FOXO4 were increased（P〈0.05）.Conclusion Use of Aplysin showed some protective effect on intestinal mucosal damage induced by alcohol exposure in rats.Its mechanism might be associated with Aplysin regulated the expression of inflammatory reaction related proteins,and reduced the release of inflammatory factor.