Effect Of Lidocaine On The Expression Of Aquaporin-5 In LPS Induced Alveolar Epithelium Type â…¡ Cells Of Rat

Objective:The aim of the study was to investigate the effect of various doses of lidocaine pretreatment on the expression of aquaporin-5 in LPS induced AT-Ⅱcells, and its possible mechanism is discussed.Methods:Primary cultured AT-Ⅱcells were randomly divided into seven groups. blank control group, Solvent control group(absolute ethyl alcohol,0.4ul/ml), LPS group, lidocainel(2μg/ml)+LPS group, lidocaine2(20μg/ml)+LPS group, lidocaine3(200μg/ml)+LPS group, lidocaine3(200μg/ml) group. The final concentration of LPS was 1μg/ml. Lidocaine was added 10 minutes before LPS. Four hours later, supernatant fluid was used to assay the content of TNF-a by ELISA, AT-Ⅱcells were used to detect the expression of aquaporin-5 mRNA and protein by reverse transcription-polymerase chain reaction and western blot.Results:1. ELISA showed that the content of TNF-a (pg/ml) in supernatant fluid of blank control group, Solvent control group and lidocaine3 group were (224.3±43.8), (222.6±34.4), (213.7±87.2) respectively. And there were no statistically significant among the three (P>0.05). Compared with the three controllers described as before, the content of TNF-a in LPS group and lidocaine+LPS groups were ascended (P<0.05). However, compared with LPS group, the release of TNF-a in lidocaine+LPS groups were decreased(P<0.05), and there were no statistically significant among lidocainel+LPS group, lidocaine2+LPS group and lidocaine3+LPS group, but they present dose dependent in data.2. RT-PCR showed that AQP-5 mRNA expressed in blank control group, Solvent control group and lidocaine3 group, which showed no statistically significant among the three (P>0.05). Compared with the three controllers described as before, the expression of AQP-5 mRNA in LPS group and lidocaine+LPS groups were descended (P<0.05). However, compared with LPS group, the expression of AQP-5 mRNA in lidocaine+LPS groups were increased(P<0.05), and there were no statistically significant among lidocainel+LPS group, lidocaine2+LPS group and lidocaine3+LPS group, but they present dose dependent in data.3. Western blot showed that AQP-5 protein expressed in blank control group, Solvent control group and lidocaine3 group, which showed no statistically significant among the three (P>0.05). Compared with the three controllers described as before, the expression of AQP-5 protein in LPS group and lidocaine+LPS groups were descended (P<0.05). However, compared with LPS group, the expression of AQP-5 protein in lidocaine+LPS groups were increased(P<0.05), and there were no statistically significant among lidocainel+LPS group, lidocaine2+LPS group and lidocaine3+LPS group, but they present dose dependent in data.Conclusion:In the cell of acute lung injury induced by LPS, 2μg/ml-200μg/ml lidocaine pretreatment can up-regulate the expression of AQP-5 mRNA and protein in AT-Ⅱcells, and the mechanism might be inhibit the release of TNF-a.