Many sedimentary metal deposits are rich in organic matter, and some organic deposits are metal deposits themselves. These metal deposits, represented by MVT Zn-Pb deposits, are widely distributed in the world, and their industrial types (Pb + Zn > 5 %) are also very rich, which has important theoretical and economic significance. However, there is no systematic research on the metallogenic mechanism of MVT Zn-Pb deposits in the participating organic matter. Therefore, geochemical simulation experiments and thermodynamic calculations have been used to examine the role of organic matter in reconstructing the ore-forming process of MVT Zn-Pb sulfide deposits. Additionally, the precipitation chemistry of sphalerite under the mineralization process mediated by organic matter is explained. Guided by the evidence on the homogenization temperature of fluid inclusion, the temperature range, from 60 °C to 200 °C, has drawn a close relationship between the temperature and the ore-forming mechanism. Notably, this reflects sphalerite’s chemical composition and morphology, even assisting in interpreting its chemical formation. The results showed that 80 °C is the beginning temperature for ZnS formation calculated from the inductively coupled plasma mass spectrometry analysis (ICP-MS) under the precipitation simulation experiments with the participation of organic matter. With the increased temperature, the rate of ZnS formation showed a trend of rapid growth in the early stage (80–140 °C) and relatively slow in the later stage (160–200 °C). This finding suggests a closer coupling relationship between metal mineralization and the maturation-genesis of organic matter, mainly the formation of the ancient oil reservoir. Following the continued increase in temperature, the morphology of ZnS was significantly different from that of the scanning electron microscope (SEM), with an apparent fine granular structure at high temperature. Likewise, more ZnO was also found through the X-ray photoelectron spectroscopy analysis (XPS), reducing the purity of the ZnS in sphalerite. Thus, this study helps reveal the metallogenic mechanism of MVT Zn-Pb sulfide deposits from the viewpoint of sphalerite chemistry under the participation of organic matter from temperature change. Furthermore, it provides a valuable theoretical and experimental basis for the later prospecting of the metal deposits.