Abstract: Objective Considering the poor teaching effect of traditional teaching mode of physiology in classroom, we studied the influence of flipped classroom teaching mode based on micro-lecture on physiology teaching, so as to further improve the teaching effect of physiology in higher vocational nursing specialty, and to provide reference for the future teaching and the study of flipped classroom teaching mode based on the micro-lecture. Methods A total of 108 students of higher vocational nursing specialty of 2016 grade from Jiangsu College of Nursing were chosen as objects of analysis. They were randomly divided into control class (54 persons) and experimental class (54 persons). Students were divided into 9 groups (6 person/group) in experimental class and practiced with flipped classroom teaching mode based on micro-lecture. Students were divided into 9 groups (6 person/group) in control class and practiced with traditional teaching mode. There were four main steps in the flipped classroom teaching mode based on micro-lecture:teacher's preparation before class; students' autonomous learning before class; seminar in class; consolidation after class. After teaching, two classes were given the same test questions in final examination, and the scores of the exam were analyzed. At the same time, the effects of teaching were evaluated according to the questionnaire survey. All data were processed by SPSS 19.0. The means of groups were compared by t test, and the enumeration data were compared by χ² test. Results The average score of the final examination of the experimental class (80.96±10.48) was significantly higher than that of the control class (71.72±13.11), and the difference was statistically significant (P<0.01). Compared with control class, the learning interest (P<0.05), learning initiative (P<0.01), knowledge understanding ability (P<0.01) and learning efficiency (P<0.01) of students were significantly improved in experimental class. Conclusion The flipped classroom teaching mode based on micro-lecture can effectively improve the teaching effect of physiology, and improve students' interest in learning, learning initiative, knowledge understanding ability and learning efficiency. Therefore, it deserves further study and promotion.