Aim To explore whether Wenyang Tongmai decoction can regulate JAK2/STAT3 signal transduction pathway and the expression of aquaporins(AQP) to protect myocardial ischemia-reperfusion tissue. Methods Thirty male SD rats were randomly divided into 5 groups, 6 in each group:sham operation group, ischemia-reperfusion group, low-dose treatment group, middle-dose treatment group, high-dose treatment group. After 14 days of intragastric administration, the model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 minutes and then releasing the ligated artery for 120 minutes. The serum levels of creatine kinase isoenzymes (CK-MB) and lactate dehydrogenase (LDH) were detected by automatic biochemical analyzer. The ST-segment elevation of the rats was counted in the electrocardiogram. Pathological changes in cardiac tissue were observed by HE staining. The expressions of AQP1 and AQP4 in myocardial tissue were detected by immunohistochemical staining. The protein expressions of JAK2, p-JAK2, STAT3, p-STAT3, AQP1 and AQP4 were detected by Western blot method. Results Compared with the sham group, the levels of CK-MB and LDH in the serum of the ischemia-reperfusion group were significantly increased (P＜0.01), and the ST-segments of the electrocardiogram were significantly elevated after ischemia for 30 minutes and reperfusion for 120 minutes(P＜0.01). HE staining showed that the myocardial cells in the ischemia-reperfusion group were severely damaged. The expressions of AQP1 and AQP4 in immunohistochemistry was significantly increased (P＜0.01). The results of Western blot showed that the expressions of JAK2, p-JAK2, STAT3, p-STAT3, AQP1 and AQP4 were significantly increased (P＜0.01). Compared with the ischemia-reperfusion group, the serum levels of CK-MB and LDH were significantly decreased in the treated rats (P＜0.01), and the ST-segments were decreased after 30 minutes ischemia and 120 minutes reperfusion (P＜0.01), myocardial cells’ damage was alleviated to varying degrees in HE staining, the protein expressions of AQP1 and AQP4 in immunohistochemistry were significantly reduced, the expressions of JAK2, p-JAK2, STAT3 and p-STAT3 were generally increased, especially the expressions of p-JAK2 and p-STAT3 (P＜0.01), AQP1 and AQP4 expression levels were reduced to varying degrees(P＜0.01). Conclusion The cardioprotective effect of Wenyang Tongmai decoction may be related to activation of JAK2/STAT3 signaling pathway and down-regulation of AQP1 and AQP4 expression.