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Results of previous studies about the acute effects of fine particulate matter (PM_2.5) on blood lipids were inconsistent. This study aimed to quantify the short-term effects of PM_2.5 on blood lipids and estimate the modifying role of insulin resistance, reflected by the homeostasis model assessment of insulin resistance (HOMA-IR). From September 2019 to January 2020, the study recruited 70 healthy adults from Wuhan University for a total of eight repeated data collections. At each visit, three consecutive days were monitored for personal exposure to PM_2.5, and then a physical examination was carried out on the fourth day. The linear mixed-effect models were operated to investigate the impact of PM_2.5 over diverse exposure windows on blood lipids. With the median of the HOMA-IR 1.820 as the cut-off point, participants were assigned to two groups for the interaction analyses. We found the overall mean level (standard deviation, SD) of PM_2.5 was 38.34 (18.33) μg/m³. Additionally, with a 10 μg/m³ rise in PM_2.5, the corresponding largest responses in triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), as well as high-density lipoprotein cholesterol (HDL-C), were −0.91% (95% confidence interval (CI): −1.63%, −0.18%), −0.33% (95% CI: −0.64%, −0.01%,), −0.94% (95% CI: −1.53%, −0.35%), and 0.67% (95% CI: 0.32%, 1.02%), respectively. The interaction analyses revealed that a significantly greater reduction in the four lipids corresponded to PM_2.5 exposure when in the group with the lower HOMA-IR (<1.820). In conclusion, short-term PM_2.5 exposure over specific time windows among healthy adults was associated with reduced TG, TC, as well as LDL-C levels, and elevated HDL-C. Additionally, the association of PM_2.5–lipids may be modulated by insulin resistance.