醋酸钠林格液复苏联合乳酸菌对创伤失血性休克大鼠肠黏膜屏障的保护作用

目的探讨醋酸钠林格液复苏基础上应用乳酸菌对创伤失血性休克大鼠肠黏膜屏障的作用。方法取30只SD大鼠数字法随机分为创伤失血性休克未复苏组（THS组，n=10），醋酸钠林格液复苏组（AR组，n=10），醋酸钠林格液联合乳酸菌复苏组（AL组，n=10），其中AL组大鼠建立休克模型前在正常喂养的基础上加服用乳酸菌素片1周（按照体积面积换算，大鼠每次服用剂量108 mg/kg，每天3次）。THS、AR组及AL组制备成休克模型平均动脉压维持（35±5）mmHg]，AL组及AR组于休克后60 min应用醋酸钠林格液进行30 min液体复苏，复苏后观察4 h取大鼠回肠组织，THS组不予复苏，于休克观察4 h后取回肠组织。利用ELISA检测大鼠外周血肿瘤坏死因子α（TNF-α）、白细胞介素（IL-6）、IL-4、及IL-10含量；应用Western blotting法检测回肠组织ZO-1、Claudin-1、TLR4、p38磷酸化及JNK磷酸化蛋白相对表达水平。结果与THS组相比，AL组大鼠外周血TNF-α、IL-6含量降低，IL-4、IL-10含量升高，AR组TNF-α含量降低，差异均有统计学意义（P<0.05）；与AR组比较，AL组大鼠外周血TNF-α含量降低，差异均有统计学意义（P<0.05），其余各细胞因子水平AR组和AL组比较差异无统计学意义（P>0.05）。回肠组织中，与THS组相比，AR组大鼠外周血TLR4、P-P38、P-JNK含量降低，AL组ZO-1、Claudin-1含量升高，差异均有统计学意义（P<0.05）；与AR组相比，AL组ZO-1、Claudin-1含量升高，TLR4、P-JNK含量降低，差异均有统计学意义（P<0.05）；回肠组织病理学结果显示AL组回肠组织损伤程度轻于THS组与AR组。结论在醋酸钠林格液复苏创伤失血性休克基础上，应用乳酸菌可能进一步抑制TLR4-p38MAPK/JNK炎性信号通路的表达，逆转休克造成的促炎因子和抗炎因子的表达失衡，减轻了创伤失血性休克肠道损伤。

Protective effect of sodium acetate Ringer's solution resuscitation combined with lactic acid bacteria on the intestinal mucosal barrier of rats with traumatic hemorrhagic shock

ObjectiveTo explore the protective effect of sodium acetate Ringer's solution resuscitation combined with lactic acid bacteria on the intestinal mucosal barrier of rats with traumatic hemorrhagic shock.MethodsThirty SD rats were randomly divided into traumatic hemorrhagic shock without resuscitation group (THS group, n=10), sodium acetate Ringer's solution resuscitation group (AR group, n=10) and sodium acetate Ringer's solution combined with lactic acid bacteria resuscitation group (AL group, n=10).The rats in the AL group were given lactic acid bacteria tablets for 1 week on the basis of normal feeding before the shock model was established (calculated according to the volume area, the rats took 108 mg/kg each time, three times a day).The shock models were established in the THS group, AR group and AL group (average arterial pressure maintained 35±5 mmHg).The AL group and AR group were treated with sodium acetate Ringer's solution for 30 minutes of fluid resuscitation after 60 minutes of shock, and the rats' ileum were observed for 4 hours after resuscitation tissue.THS group was not resuscitated, and ileum tissue was taken 4 hours after shock observation.ELISA was used to detect peripheral blood tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-4, and IL-10 concentration.Western blotting method was used to detect the expressions of ileum tissue ZO-1, claudin-1, TLR4, p38 phosphorylation and JNK phosphorylation protein.ResultsCompared with the THS group, the levels of TNF-α and IL-6 in the peripheral blood in the AL group were decreased, and the levels of IL-4 and IL-10 were increased, and the TNF-α concentration in the AR group was decreased, and the differences of which were statistically significant (P<0.05).Compared with the AR group, the level of TNF-α in peripheral blood in the AL group was decreased (P<0.05).There was no statistical difference in the levels of other cytokines between AR group and AL group (P>0.05).Compared with the THS group, the levels of TLR4, P-P38, and P-JNK in the peripheral blood of rats in the AR group were decreased, and the levels of ZO-1 and Claudin-1 were increased in the AL group, and the differences of which were statistically significant (P<0.05).Compared with the AR group, the levels of ZO-1 and Claudin-1 in the AL group were increased, and the levels of TLR4 and P-JNK were decreased (P<0.05).The histopathological results of the ileum showed that the damage of the ileum tissue in the AL group was less than that in the THS group and AR group.ConclusionsOn the basis of sodium acetate Ringer's solution to resuscitate traumatic hemorrhagic shock, the application of lactic acid bacteria may further inhibit the expressions of TLR4-p38MAPK/JNK inflammatory signal pathway and reverse the expression imbalance of pro-inflammatory factors and anti-inflammatory factors caused by shock which reduce the trauma intestinal injury in hemorrhagic shock.