Objective To investigate the influence of the pretreatment of electroacupuncture（EA） on the inflammatory cytokines and aquaporin（AQP）-5 in acute lung Injury rats induced by lipopolysaccharide. Methods SD-rats were randomly divided into 5 groups：control， ALI4h， ALI6h， EA plusALI4h and EA plus ALI6h. Acute lung injury was induced by LPS（2mg/kg） stilling into the trachea respectively for 4h and 6h except the normal group. The rats in latter two groups were pretreated with EA at bilateral Zusanli（ST36）points for a week. The plasma was separated for detecting the content of AQP5， IL-1β， IL-6， IL-10 and TNF-αby enzyme-linked immuno sorbent assay（ELISA） method. Bronchoalveolar lavage fluid（BALF） of rats was extracted for the content of AQP5 by ELISA and counting the total number of white blood cells（WBC）. The left lung tissue was used for the ratio of wet to dry weight（W/D）， and right lung was fixed in formaldehyde for observation of morphology and immunohistochemistry. Results Compared with the control group， in ALI4h group W/D and WBC count was increased（P<0.05）， the ratio of total area of alveolar to lung tissue area（A/t） was decreased（P<0.05）， IL-10， TNF-α and IL-1β were significantly increased（P<0.001）； and IL-6 and IL-10 were significantly higher（P<0.01） in ALI6h group； the content of AQP5 decreased significantly（P<0.001）， integral optical density（IOD） decreased significantly（P<0.01） in ALI4h and ALI6h groups. Compared with ALI4h group， WBC count and W/D decreased（P<0.05）， IL-1β and TNF-α content decreased（P<0.05）， AQP5 content in plasma and BALF increased（P<0.05）， and plasma AQP5 content in acupuncture group was increased in EA plus ALI4h group. Conclusion EA pretreatment at Zusanli could protect ALI rats induced LPS from inflammatory and reduce pulmonary edema， and the effect was better in ALI4h group than in ALI6h group. The mechanism may be related to the reduction of inflammatory cytokines and the up-regulation of AQP5 expression by EA pretreatment.