Carbon dioxide (CO2) flux measurements were conducted throughout the year 2022 utilizing the eddy covariance technique in this study to investigate the characteristics of carbon fluxes and their influencing factors in the Chenhu wetland, a representative subtropical lake-marsh wetland located in the middle reaches of the Yangtze River in China. The results revealed that the mean daily variation of CO2 flux during the growing season exhibited a U-shaped pattern, with measurements ranging from −12.42 to 4.28 μmolCO2·m−2·s−1. The Chenhu wetland ecosystem functions as a carbon sink throughout the growing season, subsequently transitioning to a carbon source during the non-growing season, as evidenced by observations made in 2022. The annual CO2 absorption was quantified at 21.20 molCO2·m−2, a figure that is lower than those documented for specific subtropical lake wetlands, such as Dongting Lake and Poyang Lake. However, this measurement aligns closely with the average net ecosystem exchange (NEE) reported for wetlands across Asia. The correlation between daytime CO2 flux and photosynthetically active radiation (PAR) can be accurately represented through rectangular hyperbola equations throughout the growing season. Vapor pressure deficit (VPD) acts as a constraining factor for daytime NEE, with an optimal range established between 0.5 and 1.5 kPa. Furthermore, air temperature (Ta), relative humidity (RH), and vapor pressure difference (VPD) are recognized as the principal determinants affecting NEE during the nocturnal period. The association between Ta and NEE during the non-growing season conforms to the van’t Hoff model, suggesting that NEE increases in response to elevated Ta during this timeframe.