Dyslipidemia is a metabolic disorder that can increase the risk of cardiovascular diseases. This study aims to investigate the association between serum 25- hydroxyvitamin D(25(OH)D) levels and blood lipid abnormalities in middle-aged and elderly residents of Anning District, Kunming City. It provides a theoretical basis for the prevention and treatment of cardiovascular diseases in the region. A total of 6,391 middle-aged and elderly individuals from the First People's Hospital of Anning City, Yunnan Province, were categorized based on serum 25(OH)D levels. Pearson correlation analysis, multiple linear regression, and other methods were used to examine the relationship between 25(OH)D and blood lipid abnormalities. The results showed that the Body Mass Index(BMI), Total Cholesterol(TC), and Triglyceride (TG) levels in the severely deficient group, deficient group, and insufficient group were significantly higher than those in normal group (P<0.05). Furthermore, the high-density lipoprotein cholesterol (HDL-C) levels in these groups were significantly lower than normal group (P<0.05). Correlation and regression analyses showed that serum 25(OH)D was negatively correlated with gender, age, low-density lipoprotein cholesterol(LDL-C), TC, and TG, and positively correlated with HDL-C. Taken together, these findings suggest that 25(OH)D deficiency can increase the risk of dyslipidemia; therefore, regular checkups and vitamin D supplementation are significant for the prevention of dyslipidemia.