罗哌卡因对脂多糖诱导的小鼠肺损伤的保护作用及其机制

Protective mechanism of ropivacaine on lipopolysaccharide induced lung injury in mice

目的探讨罗哌卡因对脂多糖(LPS)诱导的小鼠肺损伤的保护作用及其机制。方法30只小鼠按照随机数字表法随机分为对照组、模型组和罗哌卡因组,每组10只,模型组和罗哌卡因组大鼠经滴鼻法给予LPS,制备大鼠急性肺损伤大鼠模型,建模24 h后,罗哌卡因组大鼠经灌胃给予2 mg/kg罗哌卡因,对照组和模型组灌胃等体积生理盐水。治疗4 d后,处理大鼠,分离血清,固定肺组织。测定肺组织湿重和干重之比;苏木精-伊红(HE)染色分析两组肺组织病理学变化;酶联免疫吸附试验(ELISA)测定血清组织中白细胞介素(IL)-1β和IL-6的水平;蛋白质印迹法(Western blot)分析大鼠肺组织p65和p50蛋白表达水平。组间比较采用单因素方差分析。结果罗哌卡因组大鼠动脉血PaO_(2)[(72.96±3.52)mmHg(1 mmHg=0.133 kPa)]明显高于模型组大鼠动脉血PaO_(2)[(59.78±5.45)mmHg],差异有统计学意义(t=6.418,P<0.05)。罗哌卡因组大鼠肺组织湿重/干重比(3.16±0.16)明显低于模型组(4.68±0.38),差异有统计学意义(t=11.520,P<0.05)。罗哌卡因组大鼠肺组织病理评分[(5.05±0.57)分]明显低于模型组[(8.30±0.49)分],差异有统计学意义(t=13.640,P<0.05)。罗哌卡因组大鼠血清IL-1β和IL-6[(54.02±7.30)、(48.67±8.55)pg/ml]明显低于模型组[(128.09±12.06)、(95.36±8.68)pg/ml],差异有统计学意义(t=16.620、12.120,P<0.05)。罗哌卡因组大鼠肺组织磷酸化p38和磷酸化p65表达水平(0.45±0.05、0.38±0.05)明显低于模型组(0.72±0.08、0.62±0.07),差异有统计学意义(t=9.493、9.390,P<0.05)。结论罗哌卡因可抑制LPS诱导炎性反应,并对LPS诱导的急性肺损伤具有保护作用,可能与丝裂原活化蛋白激酶p38和核因子-κB(NF-κB)信号通路激活相关。

Objective To investigate the protective effect of ropivacaine on lipopolysaccharide(LPS)induced lung injury in mice and its molecular mechanism.Methods Thirty mice were randomly divided into control group,model group and ropivacaine group according to the method of random number table.The rats in model group and ropivacaine group were given LPS by intranasal drip to prepare the rat model of acute lung injury.After 24 hours of modeling,the rats in ropivacaine group were given 2 mg/kg ropivacaine by gavage,and the rats in control group and model group were given equal volume normal saline by gavage.After 4 days of treatment,rats were treated,serum was separated and lung tissue was fixed.The ratio of wet weight to dry weight of lung tissue was measured.The pathological changes of lung tissue in the two groups was analyzed by hematoxylin and eosin(HE)staining.The level of interleukin(IL)-1βand IL-6 in serum was analyzed by enzyme linked immunosorbent assay(ELISA).The expression level of p65 and p38 protein in rat lung tissue was analyzed by Western blotting.The measurement data were compared by one-way ANOVA.Results The arterial blood PaO_(2)[(72.96±3.52)mmHg]in ropivacaine group was significantly higher than that in model group[(59.78±5.45)mmHg(1 mmHg=0.133 kPa),t=6.418,P<0.05].The lung tissue wet weight/dry weight ratio(3.16±0.16)of rats in the ropivacaine group was significantly lower than that of rats in the model group(4.68±0.38,t=11.520,P<0.05).The lung tissue pathological score of rats in the ropivacaine group[(5.05±0.57)]was significantly lower than that of rats in the model group[(8.30±0.49),t=13.640,P<0.05].The level of interleukin-1βand IL-6 in serum[(54.02±7.30),(48.67±8.55)pg/ml]were significantly lower than serum IL-1βand IL-6 in model group[(128.09±12.06),(95.36±8.68)pg/ml,t=16.620,12.120,P<0.05].The expression levels of phosphorylated p38 and phosphorylated p65 in lung tissue of rats in ropivacaine group(0.45±0.05,0.38±0.05)were significantly lower than those of rats in model group(0.72±0.08,0.62±0.07,t=9.493,9.390,P<0.05).Conclusion Ropivacaine can inhibit LPS induced inflammation and protect LPS induced acute lung injury,which may be related to activation of mitogen activated protein kinase p38 and nuclear factor-κB(NF-κB)signaling pathway.