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Now, lodging is a major constraint factor contributing to yield loss of maize (Zea mays L.) under high planting density. Chemical regulation and nitrogen fertilizer could effectively coordinate the relationship between stem lodging and maize yield, which significantly reduce lodging and improve the grain yield. The purpose of this study was to explore the effects of chemical regulation and different nitrogen application rates on lodging characteristics, grain filling and yield of maize under high density. For this, we established a field study during 2017 and 2018 growing seasons, with three nitrogen levels of N100 (100 kg ha–1), N200 (200 kg ha–1) and N300 (300 kg ha–1) at high planting density (90000 plants ha–1), and applied plant growth regulator (Yuhuangjin, the mixture of 3％ DTA-6 and 27％ ethephon) at the 7th leaf. The results showed that chemical control increased the activities of phenylalanine ammonia-lyase (PAL), tyrosine ammonia-lyase (TAL), 4-coumarate:CoA ligase (4CL), and cinnamyl alcohol dehydrogenase (CAD), and increased the lignin, cellulose and hemicellulose contents at the bottom of the 3rd internode, which significantly reduced the lodging percentage. The lignin-related enzyme activities, lignin, cellulose and hemicellulose contents decreased with the increase of nitrogen fertilizer, which significantly increased the lodging percentage. The 200 kg ha–1 nitrogen application and chemical control increased the number, diameter, angle, volume, and dry weight of brace roots. The 200 kg ha–1 nitrogen application and chemical control significantly increased the activities of ADP-glucose pyrophosphorylase (AGPase), soluble starch synthase (SSS) and starch branching enzyme (SBE), which promoted the starch accumulation in grains. Additional, improved the maximum grain filling rate (Vmax) and mean grain filling rate (Vm), which promoted the grain filling process, significantly increased grain weight and grain number per ear, thus increased the final yield.